TABLE TO FARM: A SUSTAINABLE, SYSTEMS-BASED APPROACH FOR A SAFER AND HEALTHIER MELON SUPPLY CHAIN IN THE U.S.

• Sponsoring Institution: National Institute of Food and Agriculture
• Project Status: COMPLETE
• Funding Source: OTHER GRANTS
• Reporting Frequency: Annual
• Accession No.: 1013035
• Grant No.: 2017-51181-26834
• Cumulative Award Amt.: $4,409,547.00
• Proposal No.: 2017-03216
• Project Start Date: Sep 1, 2017
• Project End Date: Aug 31, 2024
• Grant Year: 2017
• Program Code: [SCRI]- Specialty Crop Research Initiative

Recipient Organization
TEXAS A&M UNIVERSITY
750 AGRONOMY RD STE 2701
COLLEGE STATION,TX 77843-0001

Performing Department
Vegetable & Fruit Imp. Ctr.

Non Technical Summary
This project aims to improve the safety and quality of U.S.-produced melons (Cucumis melo L.) and therefore addresses SCRI goals by: strengthening the competitiveness and sustainability of the U.S. specialty crop industry and bolstering consumption of regionally produced specialty crops, expanding the diversity of adapted varieties through breeding, and improving production practices. It is anticipated that this stakeholder-driven approach will lead to development of unique cultivars (with improved quality, safety, and sensory attributes) and increase consumer loyalty and repeat purchasing (SCRI focus areas 1, 3, and 5). The proposed project has five objectives: (1) Identify and analyze factors influencing consumer behaviors related to melon purchase and consumption; (2) Develop cultivars that address producer and consumer needs using genomics-assisted selection and breeding to harness genetic diversity of melon varieties and improve nutritional quality, flavor, safety, and stress resistance; (3) Develop region- and cultivar-specific agronomic practices and technologies to improve production efficiency, nutritional quality and safety attributes; (4) Identify critical control points and develop pre- and post-harvest strategies to minimize microbial contamination; (5) Establish a Center of Excellence Research, Extension, and Outreach Program for Melons with a focus on improving consumer awareness and meeting the consumer demand for safe, high-quality melons that exceed minimum quality standards thereby resulting in profitability for producers and retailers. The CAP team spans 13 disciplines and institutions from 7 states. We expect that enhancing melon quality, nutrition, and safety, while providing elite cultivars well-suited to regional production and distribution, will benefit producers, processors, distributors, and consumers nation-wide.

Animal Health Component

70%

Research Effort Categories

Basic

10%

Applied

70%

Developmental

20%

Classification

Knowledge Area (KA)	Subject of Investigation (SOI)	Field of Science (FOS)	Percent
201	0199	1081	100%

Knowledge Area
201 - Plant Genome, Genetics, and Genetic Mechanisms;

Subject Of Investigation
0199 - Soil and land, general;

Field Of Science
1081 - Breeding;

Keywords

flavor

food safety

health promoting properties

transdisciplinary and breeding

Goals / Objectives
The long-term goal of this multidisciplinary project is to enhance the sustainability and profitability of melon production in the U.S., emphasizing consumer preferences and industry-driven needs. This stakeholder-driven goal will be achieved by integrating market surveys, breeding, and improved production practices guided by consumer and industry expectations for retail quality, nutrition and safety. This systems approach will ensure co-optimization of cultivar development and agronomic practices to address multiple stakeholder needs.The long-term goal of this multidisciplinary project is to enhance the sustainability and profitability of melon production in the U.S., emphasizing consumer preferences and industry-driven needs. This stakeholder-driven goal will be achieved by integrating market surveys, breeding, and improved production practices guided by consumer and industry expectations for retail quality, nutrition and safety. This systems approach will ensure co-optimization of cultivar development and agronomic practices to address multiple stakeholder needs.The project goal will accomplish the five consumer-driven objectives.Objective 1. To identify and analyze factors influencing consumer behaviors related to melon purchase and consumption. We hypothesize that new melon varieties bred to be safer, healthy, and flavorful will result in higher consumer acceptance and willingness-to-pay.Objective 2. To develop cultivars/germplasm that address producer and consumer needs identified in Objective 1, using genomics-assisted selection and breeding to harness genetic diversity of melon varieties and improve nutritional quality, flavor, safety, and stress resistance. We hypothesize that genomics-assisted breeding with diverse germplasm (from TAMU, University of New Hampshire, six seed companies and CucCAP projects) will produce new cultivars with improved quality, yield, and disease resistance (see letters of collaboration in industry relevancy package).Objective 3. To develop region- and cultivar-specific agronomic practices and technologies to improve production efficiency and retail/nutritional quality and safety. We hypothesize that cultivar choice and mineral nutrient management can affect yield and consumer preference quality traits, and that crop management practices such as timing and method of irrigation affect fruit susceptibility to microbial contamination.Objective 4. To identify critical control points and develop pre-harvest (breeding and production) and post-harvest strategies to minimize microbial contamination. We hypothesize that: (i) lightly-netted genotypes (not completely smooth skin) will be less susceptible to microbial surface contamination/biofilms than densely-netted melons, (ii) susceptibility to contamination will be a function of contact with the environmental matrices, bioactive components, crop management practices and not genomic composition of the pathogen; (iii) plant antimicrobial washes, organic sanitizers, and antimicrobial edible films (in-contrast to currently used hot water or steam methods) will reduce surface contamination and microbial diversity on cantaloupes irrespective of netting; and (iv) electrostatic spray and fogging systems will be more effective than current treatments.Objective 5. To establish a Center of Excellence in Research, Extension, and Outreach for Melons with a focus on integrating all the objectives to improve consumer awareness and meeting the consumer demand for safe, high-quality melons that exceed minimum quality standards and thereby result in increased profits for producers and retailers. We hypothesize that an improved education and outreach program will raise consumer awareness and facilitate adoption of healthy cultivars, provide economic benefits to growers and improve socioeconomic impacts for all members of the supply chain (see the letters of support from industry stakeholders).

Project Methods
Following methods are used for specific objectives to accomplish the planned tasks. Method: 1.1 Assess potential melon cultivars based on their characteristics and consumer acceptance: Method 1.2 Assess the economic potential of different cultivars of melons by comparing costs of production with increased benefits. Enterprise budgets will be used to compare the production cost of selected cultivars with consumer acceptance and willingness to pay findings from Task 1.1, and how they affect the economic feasibility of melon production (Years 1-4).Method 2a. (i). Sensory evaluation: Consumer sensory testing will determine the flavor impact and acceptability of cultivars after postharvest handling and storage. Thirteen cultivars (Objective 1, Plant material) will be used for quality assessments in Year 1. Sensory panelists (n=5-10) have been selected and trained as defined by Meilgaard et al85 and have conducted descriptive attribute sensory evaluations of cantaloupe.86Method 2a. (ii). Chemo profiling of melons (flavor profile, phytochemicals and quality): Flavor profiles of melon fruit will be determined by electron impact and chemical ionization mode in GC-MS by headspace and solid phase micro extraction. The identification of individual components will be performed using our published methods.83, 84Method 3: Melons for transcriptome analysis will be sampled from the cultivars to be used for the sensory analysis. In Year 1, fruits will be collected in three biological replicates per cultivar at 35 days after pollination (DAP), which corresponds to the mid-ripening stage. Total RNA will be isolated using the RNeasy Plant Mini Kit (Qiagen, CA) 96 The mRNA purification, library construction (100-bp paired end) and RNA-seq will be carried out by TAMU Genomics and Bioinformatics Services.Method 4a. : Data integration and PCA/CCA: The data from Tasks 1-4a will be integrated to interpret and derive logical decisions on the selection from the different parameters. We will implement and evaluate other joint analysis methods, including CCA and novel low-rank factorization methods 92-95 that further consider regulations when deriving signature markers for compounds and genes that are significantly associated with customer sensory data.Method 5.1 Generate F1 hybrids by factorial mating design: We will create 125 experimental hybrids in a greenhouse using a North Carolina Design 2 mating scheme, by crossing 10 genetically diverse reticulatus females and 10 genetically diverse reticulatus males and crossing 5 by 5 with inodorus lines. Parent lines will have been selected for quality and disease resistance prior to Year 1.Method 5.2 Screen the entire set of F1 hybrids for selected agronomic traits: F1 hybrids created from Task 5.1 will be planted in a research field at Uvalde. The entire set of F1 hybrids will be evaluated for agronomic traits given in Task 9 in addition to yield, maturity date, concentrated harvest potential, and disease resistance. Harvest potential will be measured by counting the number of ripe fruits at one-week intervals.Method 6: In Year 3, the best 10 F1 hybrids will be selected based on parameters determined from data integration and PCA (Task 4a-Years 1 and 2).Method 7: Rind morphology of F1 hybrids from Task 6 will be examined. The methods are explained in Task 11a.Method 4b: Data integration and PCA/CCA: The data obtained from Tasks 2c, 3b, and 7 will be analyzed by PCA and CCA as discussed earlier for identification of signature markers associated with consumer preference. The analysis results will be provided to the breeder to enable more efficient selection based on genomic and physico-chemical properties.Methods: 8.1 Perform genome sequencing of selected germplasm and identify single nucleotide polymorphisms (SNPs): DNA will be extracted from parental lines M11-26 and M11-12 using DNeasy Plant Mini Kit (Qiagen, CA). DNA library construction (100-bp paired end) and DNA-seq will be carried out by TAMU Genomics and Bioinformatics Services. Sequence alignment and assembly will be performed using CLC Genomics Workbench (Qiagen, CA) and reads will be mapped and annotated using the reference melon genome (https://melonomics.net). SNPs and INDELs will be identified between two parents.Methods 8.2 Construct genetic populations for development of molecular markers and analyze phenotypes of progeny: Controlled pollinations to generate F1 hybrids, F2 (200 plants), F3 (200 plants), backcross (BC) generations (100 plants for BC1 and 100 plants for BC2), and new families in a greenhouse will be conducted at TAMU. Phenotypes of progeny will be determined (described in Method 5.2).Methods 8.3 Validate the markers identified from 8.1 using a genetic population and examine linkage disequilibrium: Genotyping of segregating population will be performed using SNP/INDEL markers identified from Method 8.1. Association between markers and phenotypes will be determined using the data from Method 8.2. Linkage disequilibrium will be estimated and characterized using SNP and Variation Suite v7.7.6 (Golden Helix, Inc., Bozeman, MT).Method 9: Experiments will be conducted in a randomized complete block design with four replications in each location. Agronomic data collected will include: marketable yield, plant developmental patterns (vine line, leaf area index, timing of flowering, fruit set, and fruit maturity), insect and disease damage ratings; incidence (1-5) and severity (%/plot)]. Data will be analyzed using GGEbiplot100 in which cultivars are entries and environments are testers.Method 10.1 Conduct a pathogen prevalence study in melon production districts. Methodology used for surveillance studies for Listeria and Salmonella will be as previously published, with modifications as needed.36, 101-110 Rind samples will be mixed in appropriate buffers to release pathogens, enrichments performed in appropriate selective broths and samples streaked on media specific for the target pathogens.Method 10.2 Determine lines carrying high and low microbial loads and test microbial attachment/biofims on cultivars Microbial populations on melon rinds will be analyzed on samples. The melons will be analyzed for microbial populations on the rind by total plate counts, coliform counts, and yeast and mold counts and Illumina amplicon sequencing of the 16S rRNA gene.Method 10.3: The role of soil and airborne dust particulates in cross-contamination will be studied using culture-based techniques and biophotonic imaging to visualize pathogen dispersal post contact. The melon rinds will be exposed to soil or dust inoculated with Salmonella or Listeria and the bacterial population transferred to the rind will be enumerated.Method 10.4 Evaluate postharvest sanitization procedures for bacterial intervention on elite melon hybrids: Studies will be conducted using an electrostatic spray system (dry and wet application), fog tunnel, immersion treatment, and antimicrobial edible films, to evaluate bacterial population reduction against variables of sanitizer concentration, contact time, storage time, etc.Method 10.5 Assess the correlation between pathogen prevalence, attachment, efficacy of interventions, flavor profile and disease resistance: The flavor profile of melon rinds will be analyzed by GC-MS (described in Method 2a.2). The components identified will be correlated with the foodborne disease resistance evaluation results from Tasks 10.1-10.4 to identify any signature flavor compounds that may have antimicrobial properties.

Progress 09/01/17 to 08/29/24

Outputs
Target Audience:Objective 1:The research outcomes reached a broad audience, including students, scientists, researchers, and industry stakeholders, through keynote and invited presentations at major conferences such as the ASHS, ACS (virtual), and the International Society for Horticultural Sciences in France, International Cucurbits conferences in Belgium, China, and Italy. Keynote and invited talks expanded the reach, while findings on phytochemicals and volatiles were published in the ACS Symposium series. Food label policymakers, melon wholesalers, and retailers also received this information. Sensory evaluation data were shared with global sensory professionals to enhance melon evaluation methods. The research benefits producers and seed companies and is regularly showcased at the National Center of Excellence for Melons at TAMU, which is visited by a diverse audience, including master gardeners, students, legislators, and industry professionals. Similarly, the UA Food Safety lab hosts tours and presentations on melon and other projects. Objective 2:General plant science community, particularly those interested in fruit metabolites. Multiple large melon growers in TX, AZ, NC, IA, GA, and CA, as well as several seed companies, were involved in the project to accomplish its goals. Objective 3:The project's primary audiences included >100 vegetable growers, seed companies, agrochemical representatives, postharvest stakeholders, food safety personnel, policymakers, and the scientific community. Research findings were shared through online postings, extension bulletins, peer-reviewed journals, annual Field Day events, and direct interactions with growers and industry members. Results from the 2018, 2019, and 2022 studies are available online, and additional findings have been published in international journals. The focus was on reaching consumers, growers, retailers, seed/agrochemical companies, home gardeners, and state representatives, with a specific emphasis on the melon production industry in NC. Objective 4:The target audiences for this project include melon growers, packers, processors, and consumers. Produce growers and manufacturers of natural antimicrobials have provided in-kind support to the project and have been informed about the results through seminars, oral and poster presentations, workshops, webinars, and demonstrations. Objective 5:The Melon Research and Outreach Workshops, held at the TAMU and UA, featured growers, private company representatives, faculty, and students. Presentations covered human health properties, breeding, crop management, and food safety. The workshop included demonstrations on safe melon cutting and the effectiveness of plant-based antimicrobials versus tap water for washing store-bought melons. Attendees included HRD scholars, general community members, consumers, retailers, public stakeholders, and food safety scientists. Results and outcomes were presented ton Master Gardners across the state of Texas during the state Master gardener conference. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Objective 1. The Nutrition and Quality Team comprised four postdoctoral research associates, four undergraduate students, seven graduate students, and one research assistant. The team received specialized training in phytochemicals, volatiles, postharvest practices, and quality attributes provided to a postdoctoral researcher (Singh), 7 graduate students (Ravi, Majithia, Biradar, Bhattarai, Balyan, Sah, Bastola), and a research assistant (Metrani). Additionally, one PhD student was trained in auction mechanisms and experimental design for consumer panels. Support for this team was enhanced by Ph.D. student Sixto Marquez, research associate Quishuo Song, and undergraduate students Cameron Grant and Ganga Bhatt. The sensory evaluation team involved four undergraduate students, six graduate students, and six part-time employees. Additional support came from one postdoctoral research associate, one undergraduate student, and two graduate students. Overall, the project benefited from a diverse group of researchers and students, all contributing to various aspects of training, evaluation, and project activities. Objective 2. Postdoctoral researcher Dr. Yukihiro Nagashima received training in RNA-seq analysis and data integration and participated in the analysis of melon fruit transcriptome and metabolome data. Another postdoc under Dr. Crosby's supervision learned to collectMonosporascusroot rot samples through controlled inoculations and conducted backcrosses to transfer genes into elite inbred lines of the western shipper type. A new student initiated a project to screen breeding lines for Fusarium wilt and gummy stem blight resistance. Graduate students: Nami Niyakan is analyzing RNA-seq and metabolic volatile data. Katherine Davidson received training in bioinformatics, molecular cloning, and recombinant protein expression for monoterpene synthase analysis. Kai He and Nami Niyakan developed the RNA-seq data analysis pipeline and performed differential gene expression and temporal RNA-seq analyses. Kai graduated in the Summer of 2020 and joined Qualcomm. Additionally, one postdoctoral researcher, two graduate students, one visiting PhD student, one research assistant, and two undergraduate students were trained in melon breeding, selection, quality analyses, and RNA sequencing. Objective 3. Arizona: Martin Porchas, hired as a Farm Attendant in Yuma, managed the field trial with coordination from Sonnet Nelson and oversight by Paul Brierley. He gained advanced skills in melon field trials and problem-solving, including addressing issues with fly larvae and vine decline by consulting with experts and selecting a new field site. The Senior Farm Attendant presented findings at the July 2022 Melon Research & Outreach Workshop. California: Laboratory Assistant Herman Daniel Delgado enhanced his skills in melon quality evaluation. One program technician and three technicians at UC WSREC improved their knowledge in this area. Georgia: Charlie Proudfoot, a student from the University of Georgia, contributed to the project and will present the findings at growers' meetings in winter 2020. North Carolina: The study involved six undergraduate or post-baccalaureate students and one research associate, with additional support from four full-time employees at the Central Crops Research Station. These individuals gained experience in melon development, cultivar evaluation, and data collection, potentially benefiting their future careers. Indiana: The project trained ten undergraduate students and one technician in melon production, harvest, and quality measurement. Texas-Uvalde: One undergraduate student was trained in melon field production, another in melon quality evaluation, and one graduate student collected data for their Master's thesis. Texas-Weslaco: Two undergraduate student interns and two agricultural technicians were hired and trained to assist with project activities. In summary, across multiple states, personnel were trained and gained experience in various aspects of melon research, providing valuable professional development for the participants. Objective 4. Dr. Ravishankar's lab trained one research specialist, four research technicians, seven graduate students, thirty-two undergraduate students, and seven high school students in numerous techniques and skills related to food safety research, including preparing media and buffers, dilutions, plating and enumeration techniques, sampling melons and environmental samples, measuring bacterial attachment strength, assessing pathogen transfer rate, antimicrobial treatment, biophotonic imaging, sample preparation for scanning electron microscopy, and microbiome analysis. Additionally, the research technicians, as well as graduate and undergraduate students involved in the project, were trained in oral and written communication by participating in outreach activities and preparing scientific manuscripts for publication in peer-reviewed scientific journals. In the Cooper Lab, the project trained two research technicians, two graduate students, and six undergraduate students in numerous techniques and skills related to food safety research, including microbiome studies and analysis and pathogen detection, specifically, culturing pathogens from different types of samples, collecting and processing microbiome samples from the field, DNA extraction, microbiome sequence library preparation, and microbiome data analysis. Additionally, it provided the opportunity to work and interact with different individuals in the melon industry and learn different aspects of melon production. Objective 5. The National Center for Excellence for Melons provided training opportunities to a new generation of students and scientists. Training included practical experience in melon nutrition, analytical skills, and stakeholder interactions, while others received training in food safety and field sample collection. Throughout this project, the transdisciplinary team of scientists across all participating locations trained over 40 undergraduate/post-baccalaureate students, 18 graduate students, 8 post-docs, and 2 scientists on various aspects of analytical skills, vegetable production/cultural management, diversity among melon cultivars/types, good crop husbandry practices, fruit maturity determination, and techniques for fruit quality assessments. Many of these individuals were co-authors of project publications. Some of the undergraduate and graduate students have pursued further education in specialty crop production. The research team presented results to the Advisory Board, discussing priority areas and potential impacts. How have the results been disseminated to communities of interest?Objectives 1. Dr. Patil and his team presented their research at several prestigious conferences. Patil delivered keynote presentations at International Cucurbit Conferences in Belgium, China and Italy and the American Chemical Society (ACS). His team presented at the American Society for Horticultural Sciences (ASHS) Annual Conference and the International Postharvest Conference in Wageningen. Patil's work on phytochemicals and volatile analysis was featured in the ACS symposium series. Other presentations by Patil included the Global Conference on Obesity, the 5th IUPHAR World Conference, and the Indian Horticulture Congress. Additionally, Dr. Cliff Lamb of TAMU spoke about the melon project at the Farm Bureau. Objective 2. Presentations and field demonstrations included: seed company PDs observed melons in Weslaco field plots. Student, Fix, showcased the melon breeding project with two posters at the Cucurbitaceae conference in Davis, CA, and the ASHS conference in Las Vegas. Field trials were conducted with J&D Produce and Mandujano Farms in Texas, with positive feedback from J&D and ongoing data analysis. Published research included transcriptome analysis in the ACS Book,Monosporascusresistance in Agriculture and Life Sciences Journal, melon monoterpene synthase family characterization in Molecular Horticulture, and fruit-specific transcription factors in a proceeding. Two new cantaloupe cultivars withMonosporascusresistance and high quality are set for release in HortScience (in press). Melon seeds were provided to six growers across Texas, Arizona, and California for further trials, and research was presented at the SR-ASHS Annual Conference in Oklahoma City in February 2023. Additionally, a melon field day at Weslaco was held in June 2024. Objective 3. Georgia:Results were featured at meetings throughout southwest Georgia in 2019 and 2020 by Co-PIs Coolong and da Silva. Melon trial results were featured at 4 meetings with approximately 80 total attendees during the project period. Further, one popular press article has been published. Trial results were featured on our Extension vegetable websitehttps://vegetables.caes.uga.edu/trial-results/cantaloupe.html, and recommendations were included for UGA Extension publications Cantaloupe and Specialty Melons Bulletin 1179. North Carolina:Results for North Carolina melon cultigen evaluations from 2018-2021 are available onlinehttps://rb.gy/s7uzah,https://rb.gy/0ky0g9andhttps://rb.gy/sy183c. The main audience for these studies is NC vegetable growers. Notable contributions include field visits from Seminis Seed Company representatives, and publications detailing evaluations from 2018 to 2021. Indiana:We presented results at growers' meetings, authored publications and hosted field days. We produced a video highlighting watermelon and melon variety trials conducted at the Southwest Purdue Ag Center as part of the 2021 Southwest Purdue Ag Center virtual field day, on June 24, 2021. We also developed a video highlighting specialty melon production in Indiana, for the 2021 small farm education webinar series, Aug. 2021. Texas - Weslaco:Team members-maintained dialogue with participating farmers, key stakeholders, and the Project Advisory Board Committee members, allowing the team to incorporate feedback from these groups. Our results were communicated through regular progress reports, presentations at industry meetings, field days and presentations at scientific conferences, such as the annual ASHS meetings, the ACS, and others. Several peer-reviewed publications and >5 extension bulletins were published based on data from Objective 3. We have and continue to post project results and interpretations of this project on websites frequently used by our stakeholders and consumers. In addition, the project team recently organized two field day events, (1) The Vegetable Spring Field Day at the TAMU AgriLife Research & Extension Center at Uvalde, TX, May 2024 and (2) The Melon and Cucurbit Field Day held at the TAMU AgriLife Research & Extension Center at Weslaco, TX, June 2024. Objective 4. Our results have been presented as posters at the Annual Meeting of the International Association for Food Protection (2019, 2020, and 2022), Southwest Agricultural Summit, Animal and Comparative Biomedical Sciences (ACBS) Undergraduate Poster Session, Agriculture, Life and Veterinary Sciences, and Cooperative Extension Research Poster Showcase, and Annual Food Safety Conference. The research findings were also published in peer-reviewed journals. Oral presentations of the project findings were given at the ACBS seminar series, ACS symposium series, and a Melon Workshop organized by the food safety team. Demonstrations on the use of plant-based antimicrobials and other aspects of melon food safety research were conducted by Dr. Ravishankar's team for produce industry professionals, UA alumni/donors, and local high school students. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Objective 1:We evaluated melon hybrids in diverse environments across North Carolina (NC), Texas (TX), Arizona (AZ), Indiana (IN), and Georgia (GA). These hybrids, including TX orange flesh and honeydew varieties, were compared with commercial honeydew and orange flesh melons and assessed for yield and quality in the field. Additionally, melons were sent to Texas A&M University (TAMU) for chemical and sensory evaluations; samples of the same varieties were shipped to the University of Arizona (UA) for microbial analysis, focusing on food safety. Consumer acceptance test and economic potential: We examined the value of health versus taste information and how the cost of acquiring this information (freevspaid) influences consumers' willingness to pay (WTP). The experiment, conducted with 290 participants from May to December 2022 at TAMU Human Behavior Laboratory, revealed that participants valued taste information ($1.46) significantly (p < 0.001) more than health information ($0.87). Although the cost of acquiring information did not significantly impact WTP on average, a higher WTP for taste information was associated with increased WTP for food. The study utilized a between-subject design with four conditions. Findings suggest that taste information could influence pricing strategies. Eye-tracking data corroborates that information is relevant and captures visual attention, supporting the notion that taste information might enhance overall food valuation. Quality, phytochemicals, flavor profiling, and sensory evaluation: We analyzed 164 volatile compounds in Cucumis melo samples from AZ, NC, Texas Pecos (TXP), and Texas Weslaco (TXW), including esters (crucial for ripe melon aroma), alcohols, flavor compounds like limonene and benzaldehyde, antioxidants such as β-carotene, amino acids, and total soluble solids (TSS), indicating sweetness. Bioinformatics linked metabolite profiles to sensory attributes and consumer preferences; gene expression studies focused on flavor-related monoterpene synthase genes. Consumer tests showed a strong preference for intense flavors, with F39 favored for appearance and taste and TTDV excelling overall. Sensory evaluation: Over 3 years, 6 melon and 2 honeydew varieties were planted across 7 U.S. locations (AZ, CA, NC, GA, IN, Uvalde, TX, and TXW). An expert panel evaluated whole and cut melons for sensory attributes (appearance, aroma, flavor, texture) using 16-point intensity scales and consumer preferences using 9-point hedonic scales. Whole melon appearance and aroma were poorly correlated with consumer liking for cut melons. Instead, attributes like sweetness and honey descriptors in cut melons were strongly linked to consumer preferences, whereas hard melons with musty, earthy, green, bitter, and sour notes were less favored. Juicy melons with higher surface moisture and overripe, fermented flavors were preferred. These results underscore the importance of descriptive sensory attributes in developing melon varieties with improved consumer appeal, emphasizing favorable flavor and texture traits. Objective 2:The breeding group conducted field trials for experimental melon hybrids at 5 sites in TX, 1 in AZ, 2 in NC, 1 in GA, 1 in IN, and 1 in CA, evaluating fruit quality and disease resistance. Elite hybrid seeds were produced in a College Station, TX greenhouse and through a commercial seed company in Thailand, with eight new Western shipper hybrids showing promise in TX trials. Controlled pollinations and backcrosses aimed to enhanceMonosporascus root rot resistance. Field trials at Weslaco and Uvalde, involving 34 hybrids and 28 elite inbreds, demonstrated high fruit quality, with four hybrids achieving flesh firmness over 80 N and 14 reaching a Brix of 11°. We produced 46 new F1 hybrids and evaluated 12 hybrids plus 40 elite parent lines for traits such as disease resistance and fruit characteristics, with most hybrids meeting our criteria. RNA-seq of 21 fruit samples identified ~22,000 unique transcripts and key genes related to maturation, carotenoid levels, and disease resistance. Objective 3:The project aimed to develop region-specific agronomic practices to enhance melon production efficiency and quality. We hypothesized that cultivar choice, location, and management practices affect productivity and consumer preferences, with irrigation timing and methods influencing microbial contamination. From 2018-2024, new and commercial melon cultivars were evaluated in AZ, CA, FL, GA, IN, NC, and TX under standard practices. Field trials assessed productivity and quality metrics like size, rind thickness, firmness, sweetness, and disease resistance. New cultigens from TAMU and commercial varieties were compared, with some new hybrids showing superior quality, including esters, alcohols, flavor compounds like d-limonene and benzaldehyde, antioxidants such as β-carotene, amino acids, and total soluble solids (TSS), indicating sweetness. Bioinformatics linked metabolite profiles to sensory attributes and consumer preferences, adaptability, and food safety parameters were evaluated. Data from these trials supported the development of two new hybrid cultivars, 'Supermelon' and 'Flavorific,' noted for improved disease resistance and consumer preference. Field studies in NC revealed variability in yield and quality among cultigens, with commercial varieties generally outperforming TAMU lines in shelf life and fruit size. Overall, the findings highlighted the importance of local adaptation and effective management practices for optimizing melon production and quality. Objective 4: UAtested 1,001 cantaloupes from 11 commercial fields, along with 55 soil, 55 rhizosphere, 18 water, and 33 air samples. SalmonellaandListeria were not detected in the cantaloupes or environmental samples, but cantaloupes had 1.00-3.91 log CFU/mL enterococci and 1.00-5.23 log CFU/mL coliforms. Assessing 18 melon cultivars from various regions for pathogen transfer showed that soil facilitated higher pathogen transfer than dust. Attachment studies revealed thatSalmonellaadhered more strongly to cantaloupes andListeriato hybrids. Plant-based antimicrobials, particularly oregano oil, effectively reduce pathogens. Microbiome analyses of >600 melon samples showed regional differences in bacterial diversity, with AZ melons having a lower initial microbial load than other regions. Additional testing revealed Salmonella in a few CA cantaloupes. Hybrids had higher pathogen transfer from the soil, with honeydews showing different patterns than cantaloupes. Dense netting on cantaloupes retained more bacteria, and scanning electron microscopy provided insights into fruit washing efficiency. The study also evaluated TAMU and commercially sold melon hybrids for food safety, contributing to improved melon safety practices.

Publications

• Type: Journal Articles Status: Published Year Published: 2024 Citation: Continuous flow high-pressure homogenization for preserving the nutritional quality and stability of watermelon juice under simulated market storage conditions (2024) J Adhikari, R Singh, K Adhikari, BS Patil, Innovative Food Science & Emerging Technologies, 103783.
• Type: Journal Articles Status: Published Year Published: 2024 Citation: Bacterial diversity and composition on the rinds of specific melon cultivars and hybrids from across different growing regions in the United States (2024) M Goforth, V Obergh, R Park, M Porchas, KM Crosby, JL Jifon, Plos one 19 (4), e0293861
• Type: Journal Articles Status: Published Year Published: 2024 Citation: Phthalic Acid and Its Role in Roots of Melon Plants (Cucumis melo) (2024) SA Marquez, KM Crosby, BS Patil, AMH Ibrahim, CA Avila, HP Pessoa, PhytoFrontiers" 4 (2), 121-125.
• Type: Journal Articles Status: Published Year Published: 2023 Citation: Genetic and geographical inputs that shape Metabolomic and transcriptomic profiles of melon fruits (2023) S Niyakan, Y Nagashima, J Singh, R Metrani, KM Crosby, JL Jifon, Scientia Horticulturae 321, 112337
• Type: Journal Articles Status: Published Year Published: 2024 Citation: Nondestructive assessment of maturity in cantaloupe using Raman spectroscopy with carotenoids as biomarkers (2024) GK Sah, N Goff, J Singh, KM Crosby, D Kurouski, BS Patil. Food Chemistry Advances 4, 100698.
• Type: Journal Articles Status: Published Year Published: 2023 Citation: Physicochemical, mechanical, and antimicrobial properties of sodium alginate films as carriers of zein emulsion with pelargonic acid and eugenol for active food packing (2023) BD Lade, N Dhowlaghar, SS Pillai, BS Patil Food Packaging and Shelf Life 40, 101202.
• Type: Journal Articles Status: Published Year Published: 2024 Citation: Continuous-Flow High-Pressure Homogenization of Blueberry Juice Enhances Anthocyanin and Ascorbic Acid Stability during Cold Storage (2024) J Adhikari, LR Araghi, R Singh, K Adhikari, BS Patil. Journal of Agricultural and Food Chemistry.
• Type: Journal Articles Status: Published Year Published: 2024 Citation: Lactobacillus-isolated exopolysaccharide as emulsifier ensures extended stability of eugenol encapsulation, potent anti-microbial activity, and application on fresh produce. (2024) S Balyan, V Dadwal, BS Patil Food Bioscience 61, 104632
• Type: Conference Papers and Presentations Status: Other Year Published: 2024 Citation: 2. Patil, BS., Crosby, K., Avila, C., Dadwal, V., Jha, DK., Climate Resilience and Its Influence on Bioactive Compounds in Fruits and Vegetables: Implications for Health and Gut Microbiota. Plenary presentation at the ACMAP 2024 Conference, Prairie View, Texas, USA
• Type: Conference Papers and Presentations Status: Other Year Published: 2024 Citation: Bhattarai, S., Jha, DK., and Patil, BS. Controlled environment agriculture: supplemental blue and UV-B light modulates tomato phenolic compounds and antioxidant activity in a variety-dependent manner- Oral presentation at the ACS 2024 Fall Conference, Denver USA.
• Type: Conference Papers and Presentations Status: Other Year Published: 2024 Citation: Jha, DK., Dadwal, V., Patil, BS., From Stress to Strength: Exploring Stress-induced Secondary Metabolites Production in Plants and Their Potential Impact on Gut Health Keynote presentation at the ACMAP 2024 Conference, Prairie View, Texas, USA
• Type: Conference Papers and Presentations Status: Other Year Published: 2024 Citation: Jha, DK., Singh, J., Patil, BS., Seed nanopriming with nitrogen-doped carbon dots enhances yield, quality, and phytochemicals with potential health-promoting attributes in tomato fruit. Presentation at the ACMAP 2024 Conference, Prairie View, Texas, USA
• Type: Conference Papers and Presentations Status: Other Year Published: 2024 Citation: Bhattarai, S., Jha, DK., Zhen, S., and Patil, BS. Supplemental Blue and UV-B Light Enhances Amino Acid-Derived Flavor Compounds in Greenhouse-Grown Tomatoes- Oral presentation at the ASHS 2024 Conference, Hawaii USA.
• Type: Conference Papers and Presentations Status: Other Year Published: 2024 Citation: Bhattarai, S., Jha, DK., Zhen, S., and Patil, BS. Supplemental lighting influences the antioxidant content and associated kinetic attributes in greenhouse-grown tomatoes- Oral presentation at the ACMAP 2024 Conference, Prairie View, Texas, USA.
• Type: Conference Papers and Presentations Status: Other Year Published: 2023 Citation: Balyan S., N. Dhowlaghar, and Patil, B.S., (2023). Enhancing fresh produce safety through synergistic antimicrobial activity of probiotic-derived exopolysaccharide and eugenol emulsion against foodborne pathogens. ASHS 2023 Annual Conference Orlando, Florida, USA July 31-August 4, 2023.
• Type: Conference Papers and Presentations Status: Other Year Published: 2023 Citation: Balyan S., N. Dhowlaghar, and Patil, B.S., (2023). Probiotic biopolymer-based encapsulation enhances limonene oil stability and antimicrobial efficacy. ACS Conference Fall 2023 San Francisco, California, USA August 13-17, 2023.
• Type: Conference Papers and Presentations Status: Other Year Published: 2024 Citation: Balyan S., Dadwal V., and Patil, B.S., (2024). Nanoemulsion with Lactobacillus-derived exopolysaccharide potentiates the antimicrobial activity of eugenol to protect fresh produce against foodborne pathogens and biofilms. ACS Conference Fall 2024, Denver, Colorado, USA August 1722, 2024. (Poster)
• Type: Conference Papers and Presentations Status: Other Year Published: 2024 Citation: Balyan S., Dadwal V., and Patil, B.S., (2024). Nanoemulsion with Lactobacillus-derived exopolysaccharide potentiates the antimicrobial activity of eugenol to protect fresh produce against foodborne pathogens and biofilms. ACS Conference Fall 2024, Denver, Colorado, USA August 17-22, 2024. (Presentation)
• Type: Conference Papers and Presentations Status: Other Year Published: 2024 Citation: Balyan S., Dadwal V., and Patil, B.S., (2024) Inhibiting Foodborne Pathogens and Biofilms on Fresh Produce by Nano-emulsions Developed from Lactobacillus-Derived Exopolysaccharide and Eugenol. ASHS 2024 Annual Conference Honolulu, Hawaii September 22-28, 2023
• Type: Conference Papers and Presentations Status: Other Year Published: 2024 Citation: Balyan S., Dadwal V., and Patil, B.S., (2024) Utilizing biopolymers from lactic acid bacteria to enhance the stability, bioactivity, and antimicrobial efficacy of eugenol. American Council for Medicinally Active Plants 2024 Annual Conference, Prairie View, Texas, USA October 3-6, 2024.
• Type: Conference Papers and Presentations Status: Other Year Published: 2024 Citation: Jha, DK., and Patil, B.S., (2024) Seed priming with nitrogen-doped carbon dots enhances production, reduces pungency, and improves the postharvest quality and storability of onion bulbs. ACS Conference Fall 2024, Denver, Colorado, USA August 17-22, 2024. (Presentation)
• Type: Conference Papers and Presentations Status: Other Year Published: 2023 Citation: Emulsions of Probiotic-Derived Exopolysaccharide and Eugenol to Improve the Safety of Fresh Produce Against Foodborne Pathogens (2023) S Balyan, N Dhowlaghar, BS Patil 2023 ASHS Annual Conference

Progress 09/01/22 to 08/31/23

Outputs
Target Audience:Objectives 1. Audiences were students, scientists, researchers and produce industry stakeholders. Objective 2 General plant science community, particularly those interested in fruit metabolites. Multiple large melon growers in Texas, AZ and CA and several seed companies. Objective 4 The target audiences for this project include: melon growers, packers, processors, and consumers. Produce growers and manufacturers of natural antimicrobials have been involved in this project by providing in-kind support to the project. They have been informed periodically on the results of this project through various forums such as seminars, oral and poster presentations, workshops, webinars, and demonstrations. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Objective 1. Nutrition and quality team: one Post-Doctoral Research associates, one undergraduate student worker, and two graduate students assisted with the project activities. Objective 2. A postdoc Dr. Yukihiro Nagashima has been trained for RNA-seq analysis and data integration. Another post doc under the supervision of Dr. Crosby was trained to collect Monosporascus root rot samples in the greenhouse by controlled inoculations and several backcrosses were conducted to transfer the genes into elite inbred lines of western shipper type. A new student began a project to screen the breeding lines for Fusarium wilt and gummy stem blight resistance. Objective 4. Dr. Ravishankar's lab has provided the opportunity for one research specialist, five graduate students, and two undergraduate students to learn numerous techniques and skills related to food safety research, including but not limited to sampling of melons and environmental samples for foodborne pathogens and indicator microorganisms, natural antimicrobial interventions for melon post-harvest treatment, measurement of bacterial attachment strength, assessment of transfer of pathogens, antimicrobial treatments, sample preparation for scanning electron microscopy and assessment of initial background microbiota on post-harvest melon rind surface. They were also trained in oral and written communication skills by participating in various outreach activities and preparing scientific manuscripts for publication in peer-reviewed scientifically journals. How have the results been disseminated to communities of interest?Objectives 1. Patil was also invited by the National Academies of Sciences, Washington, D.C. to give presentation. He was also invited to give Keynote presentation by International Cucurbits Conference in China (hybrid). Patil and his team members presented research outcomes as invited/ plenary/keynote speakers at the American Chemical Society, and ASHS Annual Conferences, Orlando, FL. He was also invited to give presentations at the International Postharvest conference, Wageningen, Netherland. Dr, Cliff Lamb, Director of Texas A&M University Texas AgriLife Research was invited to give presentation by the Farm Bureau and he presented about the melon project along with other AgriLife Research projects, Objective 2. Melon monoterpene synthase family characterization was published in Molecular Horticulture. Characterization of melon fruit-specific transcription factors were published in a proceeding. Release of two new cantaloupe cultivars with Monosporascus resistance and good quality, HortScience (In Press). Provided melon seed to 6 growers in Texas, Arizona and California for field trials. Presented research at SR-ASHS annual Conference in Oklahoma City, Feb 2023. Objective 4. Students, research technician and research specialist in Dr. Ravishankar's lab presented their food safety research on the melons project at the 2022 Melon Research and Outreach Workshop hosted by Dr. Ravishankar at the University of Arizona. This conference was attended by local food industry professionals, academic scientists, and the general public. Audience included more than 50 people from across the United States. The workshop also included speakers from both the University of Arizona, Yuma Center for Excellence in Desert Agriculture and Texas A&M covering a broad range of topics including growing melons, consumer and quality research and food safety research. There were also live demonstrations on washing melons, cutting melons safely and creating a melon smoothie. Dr. Cooper's graduate student presented their research results from the microbiome work. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Objective 1: Phytochemicals (volatile and non-volatile) data from GCMS and HPLC analysis, respectively were collected. Multivariate analysis was performed to determine the correlation between the phytochemicals metabolites with sensory and food safety parameters. Objective 2. The breeding group planted 5 field trials in Texas, one in Arizona and one in California with our elite, experimental melon hybrids. Fruit quality data and disease resistance data have been collected from the two early trials in south Texas. Despite excessive rainfall, several new cantaloupe hybrids and elite inbred lines produced high quality fruits with 12 °Brix or higher and firm flesh. Three trials in Texas, at Carrizo Springs and near Pecos were destroyed by hail. Seed of elite hybrids has been produced in the greenhouse at College Station and with a commercial seed company in Thailand. Seeds of 4 new western shipper hybrids were produced by controlled pollination in the greenhouse. These were included in the spring 2023 trials and hybrids 18 and 22 looked very promising in south Texas. Multiple populations were screened for Monosporascus root rot in the greenhouse by controlled inoculations and several backcrosses were conducted to transfer the genes into elite inbred lines of western shipper type. Data from these experiments were published. A new student began a project to screen the breeding lines for Fusarium wilt and gummy stem blight resistance. Objective 4. A total of 1,001 cantaloupes were harvested and analyzed from 11 commercial melon fields in Arizona, along with 55 soil, 55 rhizosphere, 18 water (10 liters each) and 33 air samples. All samples were transported to the University of Arizona for prevalence studies. Three cantaloupe melons were peeled and placed in a sterile stomacher bag to form one composite. Each composite and environmental sample was enriched in twice strength universal pre-enrichment broth, then a selective enrichment of tetrathionate broth and M-broth for Salmonella species or Listeria enrichment broth for Listeria species. Neither Salmonella or Listeria species were detected in either the melon composites or environmental samples. The countable ranges for enterococci and coliform for melon composite samples were between 1.00 - 3.91 and 1.00 - 5.23 log CFU/mL, respectively. Ranges for the countable enterococci for soil, rhizosphere, water, and air were 1.00 - 3.29, 1.00 - 3.59, 1.30 - 2.59 and 0.38 - 2.44 log CFU/mL, respectively. Ranges for the countable coliform counts were 1.00 - 3.29, 1.00 - 3.59, 1.30 - 2.59 and 0.86 - 1.26 log CFU/ml, respectively.

Publications

• Type: Journal Articles Status: Published Year Published: 2023 Citation: 13. Metrani. R., J. Singh, J., G.K. Jayaprakasha, K.M. Crosby, J.L. Jifon, S. Ravishankar, P.Brierley, D.I. Lescovar, T.A. Turini, J. Schultheis, T. Coolong, W.Guan, and B.S. Patil. 2023. Multi-location evaluation of cantaloupe (Cucumis melo L.) cultivars for their aroma and flavor related volatile composition using a metabolomics approach. Food Chemical Advances. 2: 100223.
• Type: Journal Articles Status: Published Year Published: 2023 Citation: 14. Dhowlaghar, N., T. Dhanani, S.S. Pillai, and B.S. Patil. 2023. Accelerated solvent extraction of red onion peel extract and its antimicrobial, antibiofilm, and quorum-sensing inhibition activities against Listeria monocytogenes and Chromobacterium violaceum. Food Bioscience. 53: 102649.
• Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: 15. Singh J., and Patil, B.S., (2023). A sustainable ultra-high performance liquid chromatography method for analysis of carotenoids using GRAS solvents. ACS Spring 2023, ACS Meeting & Expo, Indianapolis, March 26-30.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2023 Citation: 16. Patil, B.S. and J. Singh (2023). Chemoprofiling of common and underutilized crops to address food and nutritional insecurity in the face of climate change. ACS Fall 2023, ACS Meeting & Expo, San Francisco, Aug 13-17.
• Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: 17. Patil, B.S., J. Singh, R. Metrani, K.M. Crosby, J.L. Jifon, and S. Ravishankar Exploring the interplay between sensory attributes, food safety, bioactives, and genetics in melon cultivars: a comparative study. ISHS, VII International Symposium on Cucurbits, June 11-15.
• Type: Journal Articles Status: Published Year Published: 2023 Citation: 24. Davidson, K.H., Pillai, S.S., Nagashima, Y., Singh, J., Metrani, R., Crosby, K.M., Jifon, J., Patil, B., Niyakan, S., Qian, X., and Koiwa, H. (2023). Melon (Cucumis melo) fruit-specific monoterpene synthase. Molecular Horticulture 3, 3.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2023 Citation: 25. Patl. B.S. From the Ground Up: Unearthing the Powerful Connection Between Soil, Plant Phytochemicals, and Human Health. Invited presentation by the National Academies Sciences, Washington, D.C.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2023 Citation: 26.Ravindranath, Metrani, R., Singh, J., Jayaprakasha, G..K., Patl. B.S. A chemometric approach to investigate post-harvest behavior of Cucumis melo phytonutrients
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2023 Citation: 18. Bhattarai, S. and Patl. B.S. Supplemental Blue and UV-B Lighting Enhances Tomato Fruit Firmness and Antioxidant Compound Contents. Orlando, Florida, ASHS July 31- Aug 4.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2023 Citation: 19. Bhattarai, S. and Patl. B.S. Supplemental Blue and UV-B Lighting Enhances Volatile Organic Compounds in Greenhouse-grown Tomatoes. Orlando, Florida, ASHS July 31- Aug 4.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2023 Citation: 20. Sah, G., Crosby, K., Dadwal, V., and Patil. B.S. Influence of maturity on volatile organic compounds and amino acid composition in the rinds of different cantaloupe cultivars. Orlando, Florida, ASHS July 31- Aug 4.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2023 Citation: 21. Balyan, S., Dhowlagar, N. and Patil, B.S. Enhancing fresh produce safety through synergistic antimicrobial activity of probiotic-derived exopolysaccharide and eugenol emulsion against foodborne pathogens. Orlando, Florida, ASHS July 31- Aug 4.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2023 Citation: 22. Balyan, S., Dhowlagar, N. and Patil, B.S. Probiotic biopolymer-based encapsulation enhances limonene oil stability and antimicrobial efficacy. ACS Fall 2023, ACS Meeting & Expo, San Francisco, Aug 13-17.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2023 Citation: 23. Sah, G., Goff, N., Singh, J., Crosby, K.M.,Kurouski, D., and Patil. B.S. Nondestructive assessment of maturity in cantaloupe rind using Raman Spectroscopy with carotenoids as biomarker. ACS Spring 2023, ACS Meeting & Expo, Indianapolis, March 26-30.
• Type: Journal Articles Status: Published Year Published: 2023 Citation: 1. Marquez, S., Jifon, J., Crosby, K., Avila, C., and A. Ibrahim. 2023. Heterosis of Vine Decline Disease Resistance Caused by the Fungus Monosporascus
• Type: Journal Articles Status: Published Year Published: 2023 Citation: 2. Marquez, SA., Crosby, K., Patil, B., Avila, C., Ibrahim, AMH., Pessoa, H and Singh, J. 2023. Hydroxy proline and gamma aminobutyric acid: markers of susceptibility to vine decline disease caused by the fungus Monosporascus cannonballus in melons (Cucumis melo L). PeerJ 11: e14932 http://doi.org/10.7717/peerj.1432.
• Type: Other Status: Other Year Published: 2023 Citation: 3. Crosby, K., Marquez, S., Jifon, J., Leskovar, D., Singh, J., Isakeit, T., and B. Patil.2023. Supermelon and Flavorific: two new hybrid muskmelon cultivars with resistance to Monosporascus cannonballus from Texas A&M AgriLife Research. HortScience: In press.
• Type: Other Status: Other Year Published: 2021 Citation: 4. Crosby, K., Marquez, S., Leskovar, D., Jifon, J., Singh, J., and B. Patil. 2023. New melon cultivars from Texas A&M AgriLife Research. HortScience: In Press.
• Type: Conference Papers and Presentations Status: Other Year Published: 2022 Citation: 6. Rowlands, D., Raygoza, K., Bains, K., Porchas, M., Brierley, P., Crosby, K., Patil, B. and Ravishankar, S. Attachment Strength of Salmonella enterica serovar Newport and Listeria monocytogenes on Melon Hybrids from Five Growing Regions in the United States. Poster presentation given at the Annual Meeting of the International Association for Food Protection, Pittsburgh, PA, August 1, 2022.
• Type: Conference Papers and Presentations Status: Other Year Published: 2022 Citation: 7. Ravishankar, S. Emerging Foodborne Microbial Threats: Challenges and Control Strategies. Invited virtual oral presentation at the International Microorganism Day  2022 as part of an International Webinar on The Importance of Microorganisms in Food Safety, hosted by the Sathyabama Institute of Science & Technology, India, September 23, 2022
• Type: Other Status: Other Year Published: 2022 Citation: 8. Park, R., Why We Peel Melons: A Peek into the Prevalence of Foodborne Pathogens Among Field-Grown Melons in Arizona and Environmental Risk Factors for Cross-Contamination. International Webinar on The Importance of Microorganisms in Food Safety, Virtual, October 2022.
• Type: Conference Papers and Presentations Status: Other Year Published: 2022 Citation: 9. Rowlands, D., Raygoza, K., Bains, K., Porchas, M., Brierley, P., Crosby, K., Patil, B., and Ravishankar, S. Attachment Strength of Salmonella enterica serovar Newport and Listeria monocytogenes on Melon Hybrids from Five Growing Regions in the United States. Annual Food Safety Conference, Tucson, AZ, October 2022.
• Type: Conference Papers and Presentations Status: Other Year Published: 2022 Citation: 10. Ravishankar, S. Emerging Foodborne Microbial Threats: Challenges & Control Strategies. Invited virtual oral presentation at the 2022 Arizona Virtual Annual Sanitarians Conference, November 2, 2022.
• Type: Conference Papers and Presentations Status: Other Year Published: 2022 Citation: 5. Park, R., Rowlands, D., Porchas, M., Brierley, P., Patil, B., and Ravishankar, S. Cross-Contamination from Environmental Matrices: A Vehicle for Transfer of Foodborne Pathogens to Melons Grown in Various Regions of the United States. International Association for Food Protection Annual Conference, Pittsburgh, Pennsylvania July-August 2022.
• Type: Conference Papers and Presentations Status: Other Year Published: 2022 Citation: 11. Ravishankar, S. Emerging Foodborne Microbial Threats in Produce: Challenges & Control Strategies. Invited virtual oral presentation at the 2022 Maritime Silk Road International Conference on the Cooperation and Integration of Industry, Education, Research and Application, hosted by the Fujian Agriculture & Forestry University, Fuzhou, China, November 22, 2022.
• Type: Conference Papers and Presentations Status: Other Year Published: 2022 Citation: 12. M. Goforth, V. Obergh, R. Park, S. Ravishankar, C.T. Parker, and K.K. Cooper. Microbial diversity of different melon types grown in different regions in the United States (Poster), International Food Protection Association Annual Meeting, Pittsburgh, PA (July).
• Type: Theses/Dissertations Status: Published Year Published: 2023 Citation: 13. M. Goforth. (2023). Characterization of fruit carposphere within the United States, masters thesis. The University of Arizona, Tucson, AZ.

Progress 09/01/21 to 08/31/22

Outputs
Target Audience:Objective 1. The outcomes of the research were disseminated to a global audience through three professional conferences: American Society for Horticultural Sciences, Chicago, IL; American Chemical Society (virtual); and the International Society for Horticultural Sciences, Angers, France. Being keynote/invited presentations helped to cover a larger audience. The phytochemicals and volatile analysis output has been published in the American Chemical SocietySymposium series. Food label policymakers, melon wholesalers, and retailers also received the information. Objective 2. The general plant science community, particularly those interested in fruit metabolites. Multiple large melon growers in Texas and several seed companies Objective 3. Consumers, growers, retailers, seed/agrochemical companies, the scientific community, home gardeners, and state (Texas Department of Agriculture) representatives. North Carolina: Target audience to be reached is the industry in melon production. Objective 4. The target audiences for this project include melon growers, packers, processors, and consumers. Melon growers and manufacturers of natural antimicrobials have been involved in this project by providing in-kind support to the project. They have been informed periodically about the results of this project through various forums such as seminars, oral and poster presentations, workshops, webinars, and demonstrations. We have presented the results to numerous members of the produce industry, federal regulatory agency representatives, academic researchers, and federal government researchers, which has allowed us to discuss the results of the studies with them in detail and address any questions that they had about the results. Objective 5.The hybrid Melon Research and outreach workshop was arranged at the University of Arizona, Tucson. Participants include growers, private company representatives, faculty, and students. Researchers and students presented their research related to human health properties, breeding, crop management, and food safety. The workshop also included demonstrations of 'how to cut melons safely' as well as the 'Effectiveness of Washing your Melon from the Store Using Plant-based Antimicrobials vs. Tap Water. Presented outcomes of the research to local food industries as a part of the Food Safety Consortium at UA on May 18, 2022. Participated in Yuma Fresh Veg Association Southwest Ag summit 2022 and presented research outcomes by the food safety team. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Objective 1 Nutrition and quality team: Four post-doctoral research associates, one undergraduate student worker, five graduate students, and one research assistant assisted with the project activities. One Ph.D. student (Sixto Marquez), one research associate (Quishuo Song), and two undergraduate students (Cameron Grant and Ganga Bhatt) also assisted with the project activities. Sensory team: part-time employees participated in the sensory evaluation study. Objective 2 A graduate student (Nami Niyakan) has been developing the RNA-seq and metabolic volatile data analysis and performed condition-dependent differential expression and association analyses for both data types. A graduate student (Katherine Davidson) has been trained in bioinformatics, molecular cloning, and recombinant protein expression for monoterpene synthase analysis. A postdoc (Dr. Yukihiro Nagashima) has been trained for RNA-seq analysis and data integration. Objective 3 Arizona - Martin Porchas, YCEDA Sr. Farm Attendant, presented information on the Yuma melon trials at the July 2022 Melon Research & Outreach Workshop with Dr. Ravishankar's research staff. North Carolina - This study provided the opportunity to engage and teach 4 recently graduated undergraduates, two undergraduate students, two graduate students, and a research associate. These employees became familiar with general melon development over the season, experienced and learned the differences between the various hybrids and cultivars, and learned the importance and how to take various quality measures such as soluble solids and flesh firmness, as well as experimental setup and its importance in conducting field research. All four graduated undergraduate students have found successful employment. Texas-Weslaco: Three undergraduate student interns and two ag technicians were hired/trained to assist with project activities. Texas-Uvalde: Two undergraduate student interns and one technician were trained/hired (partial funds) to assist with project activities. Objective 4 Food safety team: one research specialist, one research technician, two graduate students, and seven undergraduate students to learn numerous techniques and skills related to food safety research, including sampling of melons and environmental samples for foodborne pathogens and indicator microorganisms, natural antimicrobial interventions for melon post-harvest treatment, measurement of bacterial attachment strength, assessment of transfer of foodborne pathogens, antimicrobial treatments, sample preparation for scanning electron microscopy and assessment of initial background microbiota on post-harvest melon rind surface. They were also trained in oral and writing communication skills by participating in various outreach activities and preparing scientific manuscripts for publication in peer-reviewed scientifical journals. In the Cooper lab, two new graduate students and six undergraduate students were trained on the proper techniques for collecting microbiome samples, processing the samples, DNA extraction microbiome sequence library preparation, and microbiome data analysis. How have the results been disseminated to communities of interest? Objective 1. Dr. Bhimu Patil and his team members presented research outcomes as invited/ plenary/keynote speakers at the American Society for Horticultural Sciences Annual Conference as well as Agriculture and Food Chemistry of the American Chemical Society, International Society for Horticultural Sciences. The phytochemicals ad volatile analysis output has been published in the American Chemical Society symposium series. Dr. Marco's team's research outcome will help policymakers and promotion boards to prescribe some guidelines for the selection between health and tasteinformationon the label of food products and to be aware of the impact of costlyinformationon food choices. Objective 2. The transcriptome analysis has been published in the American Chemical Society Book. Publication of Monosporascus resistance in Agriculture and Life Sciences Journal. Provided seed to four growers in TX and CA for field trials. Objective 3. The results of this research will be made available in an online extension publication that will be published in the next several weeks. Objective 4. Food safety team members had the opportunity to participate in the poster session at the 2022 Southwest Agricultural Summit held in Yuma, AZ, to provide information to the stakeholders and public on melon safety issues. Demonstrations of the use of plant antimicrobials and other aspects of melon food safety research were conducted for produce industry professionals, members of the scientific community, and high school students throughout the year. Objective 5 .We disseminated information to the scientific community, industry, and the public through a workshop at the University of Arizona in Tucson, AZ in July 2022. The results from the Cooper lab have been disseminated through poster presentations at two food safety conferences, discussions directly with individuals in the produce industry, and an invited oral presentation to numerous researchers at the USDA, ARS, WRRC in Albany, CA.Results were shared with melon growers and other producers through the respective state growers' associations meetings and conferences. Presentations to Highschool students as part of USDA Ag Discovery Camp- June 2022 and SafeFoodCats Highschool students camp on June 10, 2022 at University of Arizona. Documnetary was prepared on Dr. Ravishanker and their researchby Arizona Public Media on Aug 11, 16 and 24, 2022. What do you plan to do during the next reporting period to accomplish the goals?Objective 1.All the phytochemical analysis for melons grown collaborators states be conducted.sensory data will be analyzed and establish a correlation among all the parameters including quality. . The statical analysis will be performed to interpret outcomes. Objective 2.We will finalize the metabolite expression analysis pipeline, including the customized imputation, differential expression, and condition-dependent profile analyses to help identify the metabolites uniquely enriched/absent in each cultivar/location investigated in this project. The prioritized metabolites will be further investigated to identify associated genes and/or pathways as potential genetic markers under different conditions by joint analysis of metabolite and RNA-seq expression, and sensory data. Objective 3.Texas Uvalde, and Weslaco: Yield and quality data from all the previous trials will be pooled and analyzed to identify lines with consistently superior performance under local production conditions. North Carolina: As planned, a melon field study in NC was conducted in 2021. Yields and quality attributes were determined. Shipped fruits for internal chemistry and sensory analysis to further characterize the effects of environment/location on these attributes. Plans are to complete and post the results of this study online. Data from all trial sites (AZ, NC, and TX) will also be subjected to a combined analysis to identify lines that are suitable for each production region. The results of this research will be made available in on-line extension publication that will be published in the next several weeks. Objective 4. We plan to complete publications using the data collected from the project in the labs of Dr. Ravishankar and Dr. Cooper. Food safety team members are working on publications using the data collected. Four publications from the research conducted by the Cooper laboratory are in preparation. This includes a co-publication with the Ravishankar laboratory (currently being written) and three different microbiome publications. Objective 5 .We plan to conduct an in-person stakeholder engagement conference. Log-Model of all inputs, short term and long term goals will be compiled.

Impacts
What was accomplished under these goals? Objective 1. We identified the characteristic impact flavor or aroma compounds (CIFAC) in Cucumis melo, with 164 volatile compound profiles generated across melon samples from Arizona (AZ), North Carolina (NC), Texas Pecos (TXP), and Texas Weslaco (TXW). The total esters that contribute to the aroma of ripe melons were the highest in TH-18 from NC and the lowest in TH-13 from TXW. The individual melon esters ethyl butanoate, ethyl 2-methyl butanoate, and ethyl hexanoate were the highest in TH-10 from AZ and benzyl acetate was the highest in TH-1 from TXP. The total alcohols were the highest in TH-6 from TXP and lowest in F39 from AZ where (Z)-6-nonen-1-ol was highest in HTIG and TH-17 from AZ. The aldehydes were found to be highest in TH-21 from AZ and lowest in NUN from TXP, among which (Z)-6-nonenal was highest in TH-17 from AZ, (E)-2-nonenal and (E, Z)-2,6-nonadienal were highest in TH-12 from AZ. Additionally, limonene and benzaldehyde, which impart flavor and aroma, and possess antimicrobial properties, were also found in melon samples with the highest limonene measured in TTDV from TXW and the highest benzaldehyde measured in TH-9 from AZ. β-Carotene was significantly higher in the selected melon hybrids and cultivars (TH-5, TH-6, and TH-9) grown in TXP and AZ. Total soluble solids (TSS, °Brix) measure the sweetness and help in determining the ripeness of fruits. The hybrids TH-6, TH-9, TH-11, TH-13, TH-16, and TH-20 grown in different locations showed higher TSS contents than other cultivars. Using bioinformatics approaches, metabolites uniquely enriched/absent in each cultivar/location are being identified. The relationship of individual metabolites with sensory parameters is being evaluated to identify factors that determine consumer preference. Gene expression profiles are being analyzed to determine the suites of transcripts supporting the flavor profile. Nine monoterpene synthase genes were analyzed and isoforms responsible for terpineol and eugenol synthesis were identified. We are using eye-tracker technology in a laboratory experiment with grocery shoppers (nonstudents) to further decode the importance of information acquisition and how it impacts the willingness to pay for melons. The current study is at the data collecting stage. We expect to recruit a total of 240 participants from the general population in Bryan and College Station areas to complete an experiment conducted at the Human Behavior Lab at Texas A&M University. Until now, we have collected responses from 163 participants. Objective 2. The breeding group planted and evaluated three field trials in Texas and one in California with our elite, experimental melon hybrids. Fruit quality and disease resistance data have been collected for publication. Seed for the four best hybrids has been produced in the greenhouse at College Station and with a commercial seed company in Thailand. Seeds of eight new Western Shipper hybrids were produced by controlled pollination in the greenhouse. These were included in the spring 2022 trials and two looked very promising in South Texas. Multiple populations were screened for Monosporascus root rot in the greenhouse by controlled inoculations and several backcrosses were conducted to transfer the genes into elite inbred lines of Western Shipper type. Data from these experiments were published. We have developed bioinformatics analysis pipelines, particularly condition-dependent differential expression and association analyses, for both RNA-seq and metabolite expression data for melon fruit of different types and locations. These analyses aim to identify potential genetic markers for metabolites of interest, particularly those related to fruit characteristics directly affecting melon supply and market. Objective 3. During the spring 2022 growing season, eight new breeding lines and new hybrid melon genotypes were grown in TXW. A limited number of commercial melon genotypes were previously evaluated in 2021 and parent lines of the new hybrids were also evaluated. Melon varieties 'Infinite Gold', a commercial Harper-type, Western Shipper, and 12 breeding lines (TH-5, TH-6, TH-9, TH-12, TH-13, TH-14, TH-16, TH-17, TH-18, TH-19, TH-20, and TH-22) were grown in TX-Uvalde. Fruit samples from these field trials were analyzed for retail, and quality traits. Fourteen melon hybrids were evaluated for yield, fruit size, and internal qualities in NC. Melon samples were shipped for phytochemical analysis to College Station, TX, and food safety evaluation to Tucson, AZ. Commercial melons were harvested from two different fields in the Yuma, AZ area. The melon samples were delivered to Dr. Ravishankar's food safety lab, AZ, for analysis along with corresponding environmental samples (air, water, soil, rhizosphere) from the harvested fields. Objective 4 . Different melon cultivars (TH-1, TH-5, TH-6, TH-9, TH-10, TH-12, TH-13, TH-14, TH-16, TH-17, TH-18, TH-19, TH-20, TH-21, F-39, Davinci SAKATA, Infinite Gold, and Harper Type from four growing locations (AZ, TX, North Carolina [NC], and California [CA]) were received for food safety studies. Melon rinds were cut into rectangular pieces and inoculated with Salmonella enterica or Listeria monocytogenes in soil and dust. Percent transfer of L. monocytogenes was highest for soil-inoculated NC TH17 and AZ TH6, and lowest for TX Infinite Gold, TX TH6, and CA TH6. Percent transfer of Salmonella was highest for soil-inoculated AZ TH19 and lowest for NC TH18. The highest percent transfer from dust for L. monocytogenes was AZ F39 and the lowest for AZ TH21. For Salmonella the highest transfer from dust was for AZ TH20 and lowest for TX Infinite Gold. Overall, the soil had a higher percent transfer for Salmonella and L. monocytogenes than dust. In general, Salmonella had a higher percent transfer than Listeria from soil and dust. The attachment strength of Listeria monocytogenes and Salmonella Newport on melon hybrids were evaluated. The strongest average attachment strength for Salmonella was on TH12 and the lowest was on TH6. The strongest average attachment strength for L. monocytogenes was on TH21, while the lowest was on Harper MegaPunk. In general, the attachment strength of Salmonella was slightly higher on the cantaloupe varieties than the hybrids, while L. monocytogenes was higher in the hybrids than the cantaloupe varieties. Melon rinds were immersed in a plant-based antimicrobial solution of apple extract or oregano oil to study their antimicrobial activity. Phosphate-buffered saline (PBS) was used as a control. The plant-based antimicrobials reduced S. Newport and L. monocytogenes populations on all samples, regardless of the melon types, varieties, or growing locations. In most cases, plant-based antimicrobial treatments reduced the pathogen population to below the detection limit (1 CFU/g) at day 3. In general, oregano oil had better antimicrobial activity than apple extract. Different microbiome samples collected from different melons were analyzed and the results showed that there was a statistically significant difference in bacterial diversity based on the type of melons, but no significant difference in bacterial diversity based on a region of growth.

Publications

• Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Park, R., Rao, A., Rowlands, D., Wei, Q., Brierley, P., Porchas, M., Patil, B. and Ravishankar S. (2021) Why We Peel Melons: A Peek into the Prevalence of Foodborne Pathogens Among Field-Grown Melons in Arizona. Oral presentation given at the Annual Food Safety Conference, Tucson, AZ, October 21, 2021.
• Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Alshahrani, S., Getchell, K., Raygoza, K., Bains, K., Murell, V., Brierley, P., Porchas, M., Patil, B. and Ravishankar, S. (2021) A Comparison of Initial Background Microbiota on Surface of Melons Grown in Different Regions of the United States. Poster presentation given at the Annual Food Safety Conference, Tucson, AZ, October 20, 2021.
• Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Ravishankar, S. 2022. Emerging Foodborne Microbial Threats: Challenges & Control Strategies. Virtual presentation given at the Microbiology Consortium of the Philippines and the Philippine Society for Microbiology Spring Webinar Series, April 29, 2022.
• Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Ravishankar, S. 2022. Current Trends in Biotechnological Interventions in Food Safety. Invited oral presentation given at the Amity University, Mumbai, India on May 31, 2022.
• Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Goforth, M., Obergh, V., Park, R., Ravishankar, S., and Cooper K.K. (2021). Microbial diversity of different melon types grown in different regions in the United States. UArizona Food Safety Conference. Oct. 2021.
• Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Cooper, K.K. (2022). Characterizing the microbiome of fruits and vegetables to address food safety issues. Invited talk at USDA, ARS, WRRC in Albany, CA. May 2022.
• Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Goforth, M., Obergh, V., Park, R., Ravishankar, S., and Cooper K.K. (2022). Microbial diversity of different melon types grown in different regions in the United States. International Association of Food Protection Annual Meeting. Aug 2022.
• Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Van, H. T. M., Nafukho, F. M., & Patil, B. (2021). Application of the logic models of evaluation to multidisciplinary research  The case of a federally funded project. Proceedings of the 2021 Academy of Human Resource Development Conference. Academy of Human Resource Development International Research Conference. St. Paul, MN: Academy of Human Resource Development. 2021.
• Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Jia, R., Palma, M.A., Bryant, H., Fischer, B. (2022). Costly information acquisition and WTP for melon Agricultural and Applied Economics Association(AAEA) Annual Conference Aug. 2022
• Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Singh J., Metrani, R., Crosby, K.M., Jifon, J., Ravishankar, S., Paul, B., Leskovar D., Guan, W., Miller, R., Patil, B.S., (2022). Sensory and Phytochemical Attributes of Muskmelon Cultivars as a Function of Production Locations. ASHS Annual Conference Chicago, July 30 to Aug 3, 2022.
• Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Singh J., Metrani, R., Crosby, K.M., Jifon, J., Miller, R., Patil, B.S., (2022) Variation in Sensory Traits And Phytochemical Composition Among Muskmelon Cultigens Grown In Semi-Arid Conditions In South Texas. ASHS Annual Conference Chicago, July 30 to Aug 3, 2022.
• Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Crosby,K., Jifon,J., Marquez, S., Leskovar, D., Grubbs, H., and B. Patil. 2021 Developing High Quality Muskmelon Hybrids with Monosporascus cannonballus Resistance. HortScience 56 (9): S126.
• Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Marquez, S., Marquez, S.,Crosby,K., Avila, C., Patil, B., and A. Ibrahim. 2021. Generation Means Analysis of Vine Decline Resistance in Melons (Cucumis melo L.). HortScience 56 (9): S104.
• Type: Journal Articles Status: Accepted Year Published: 2022 Citation: . M�rquez, S., K, Crosby., C, Avila., B, Patil and A, Ibrahim. 2022. Generation means analysis of vine decline disease resistance in melons (Cucumis melo L). Agriculture and life sciences. In press.
• Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Rowlands, D., Raygoza, K., Brierley, P., Porchas M., Patil, B. and Ravishankar, S. (2021) Attachment Strength of Salmonella enterica serovar Newport and Listeria monocytogenes on Melon Hybrids Grown in Five Different Regions of the United States. Poster presentation given at the Annual Food Safety Conference, Tucson, AZ, October 20, 2021.
• Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Alshahrani, S., Getchell, K., Raygoza, K., Bains, K., Murrell V., Brierley, P., Porchas, M., Patil, B. and Ravishankar S. (2022). A Comparison of Initial Background Microbiota on the Surface of Melons Grown in Different Regions in the United States. Poster presentation given at the Southwest Agriculture Summit 2022 Student Poster Session, Yuma, AZ, February 23, 2022.
• Type: Book Chapters Status: Published Year Published: 2022 Citation: Manchali, S., Murthy, K.N.C., Patil. B.S. 2022, Native Dietary Plant Crops: Treasure House of Nutrition and Bioactive Molecules, Chapter 4 In Patil, B.S., Dhowlaghar, N., Singh, J. and Pillai, S. Melon breeding and Genetics. ACS Symposium Series 1415 DOI: 10.1021/bk-2022-1415.ch002. p. 55-66.
• Type: Journal Articles Status: Published Year Published: 2021 Citation: Manchali, S., Murthy, K.N.C., Vishnuvardana, Patil. B.S. 2021. Nutritional Composition and Health Benefits of Various Botanical Types of Melon (Cucumis melo L.). Plants 2021, 10(9), 1755.
• Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Park, R., Rowlands, D., Porchas, M., Brierley, P., Crosby, K., Patil, B., and Ravishankar, S. Cross-contamination from environmental matrices: A vehicle for transfer of foodborne pathogens to melons grown in various regions in the United States. International Association for Food Protection Annual Meeting. Aug 2022
• Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Rowlands, D., Raygoza, K., Bains, K., Porchas, M., Brierley, P., Crosby, K., Patil, B., and Ravishankar, S. Attachment strength of Salmonella enterica serovar Newport and Listeria monocytogenes on melon hybrids from five growing regions in the United States. International Association for Food Protection Annual meeting. Aug 2022.
• Type: Journal Articles Status: Published Year Published: 2022 Citation: Metrani. R., Jayaprakasha, G.K., and Patil., B.S. Optimization of Experimental Parameters and Chemometrics Approach to Identify Potential Volatile Markers in Seven Cucumis melo Varieties Using HSSPMEGCMS. Food Analytical Methods, 15 (3), 607-624.
• Type: Journal Articles Status: Published Year Published: 2022 Citation: Zhu, L., Wei, Q., Porchas, M., Brierley, P., Friedman, M., Crosby, K., Patil, B., and Ravishankar, S. 2022. Plant-Based Antimicrobials Inactivate Listeria monocytogenes and Salmonella enterica on Melons Grown in Different Regions of the United States. Food Microbiology 101:103876.
• Type: Journal Articles Status: Published Year Published: 2021 Citation: Singh, J., R. Metrani, G.K. Jayaprakasha, K.M. Crosby, J.L. Jifon, S. Ravishankar, P.Brierley, D.L. Lescovar, T.A. Turini, J. Schultheis, T. Coolong, W.Guan, and B.S. Patil. 2022. Profiling carotenoid and sugar contents in unique Cucumis melon L. Cultigens harvested from different climatic regions of the United States. J. Food Composition and Analysis. 106:1-8
• Type: Journal Articles Status: Published Year Published: 2022 Citation: Birader. K., Singh, J., Pillai, SyamKumar S., and Patil. B.S.. 2022. Separation of nordihydrocapsiate from capsiate and major capsaicinoid analogues using ultra high performance liquid chromatography. Food Chemistry, 382, 132585.
• Type: Journal Articles Status: Published Year Published: 2021 Citation: Majithia D., Metrani. R., Dhowlaghar. N., Crosby. K.S., and Patil. B.S. Assessment and classification of volatile profiles in melon breeding lines using headspace solid-phase microextraction coupled with gas chromatography-mass spectrometry. Plants, 10 (10), 2166.
• Type: Journal Articles Status: Published Year Published: 2021 Citation: Acharaya P., Singh J.S., Jayaprakasha G.K., Jifon, J.L., Crosby, K.M., Paitl, B.S. Impact of storage period and nanoparticle treatment on phytochemical composition of watermelons (Citrullus lanatus). Journal of Food Composition and analysis, 104, 104139.
• Type: Book Chapters Status: Published Year Published: 2022 Citation: Adhikari. J., Singh, J., Patil. B.S. 2022. Influence of Processing and Storage on the Blueberry Phytonutrients, Chapter 8 In Patil, B.S., Dhowlaghar, N., Singh, J. and Pillai, S. Melon breeding and Genetics. ACS Symposium Series 1415 DOI: 10.1021/bk-2022-1415.ch002.p. 117-126.
• Type: Book Chapters Status: Published Year Published: 2022 Citation: Singh, J., Metrani. R. Patil, B.S. Effect of Locations on Phytonutrients Contents in Muskmelons: A Review, Chapter 2 In Patil, B.S., Dhowlaghar, N., Singh, J. and Pillai, S. Melon breeding and Genetics. ACS Symposium Series 1415 DOI: 10.1021/bk-2022-1415.ch002. p. 23-36.
• Type: Book Chapters Status: Published Year Published: 2022 Citation: Metrani. R, Singh, J., Dhowlaghar, N., Patil. B.S. Profiling of Volatile Compounds in Melons and Their Implication on Flavor, Aroma, Quality, and Food Safety, Chapter 3 In Patil, B.S., Dhowlaghar, N., Singh, J. and Pillai, S. Melon breeding and Genetics. ACS Symposium Series 1415 DOI: 10.1021/bk-2022-1415.ch002. p. 37-54.
• Type: Book Chapters Status: Published Year Published: 2022 Citation: Nagashima, Y., Niyakan,S., He, K., Singh, J., Metrani, R., Crosby, K.M., Jifon, J., Jayaprakasha, G.K., Patil, B.S., Qian, X., Koiwa, H. Differential Expression of Transcription Factors, Chapter 1 In Patil, B.S., Dhowlaghar, N., Singh, J. and Pillai, S. Melon breeding and Genetics. ACS Symposium Series 1415 DOI: 10.1021/bk-2022-1415.ch002.p. 3-21.
• Type: Book Chapters Status: Published Year Published: 2022 Citation: Park, R., Rowlands, D., Wei, Q., Brierley, P., Porchas, M., Crosby, K., Patil, B., Friedman, M., and Ravishankar S. Understanding Cross-Contamination of Listeria monocytogenes and Salmonella enterica via Environmental Matrices onto melons Grown in Different Regions of the United States, Chapter 5, In Patil, B.S., Dhowlaghar, N., Singh, J. and Pillai, S. Melon breeding and Genetics. ACS Symposium Series 1415 DOI: 10.1021/bk-2022-1415.ch002. p.69-87.
• Type: Book Chapters Status: Published Year Published: 2022 Citation: Dhowlaghar, N. and Patil, B.S. 2022, Factors Affecting Foodborne Pathogen Contamination in Melons during Production and Their Management, Chapter 6, In Patil, B.S., Dhowlaghar, N., Singh, J. and Pillai, S. Melon breeding and Genetics. ACS Symposium Series 1415 DOI: 10.1021/bk-2022-1415.ch002. p. 89-97.

Progress 09/01/20 to 08/31/21

Outputs
Target Audience:Objective 1: Due to the COVID-19 pandemic, the center had no visitors. But the results were presented at various virtual conferences, 2021 ASHS Annual Conference, ACS National Meeting & Exposition, III Asian Horticultural Congress - AHC2020. Objective 2: General plant science community. Objective 3: North Carolina: Professionals at the ACS conference. Texas - Weslaco: Consumers, growers, retailers, seed/agrochemical companies, the scientific community, home gardeners, and state (Texas Department of Agriculture) representatives. Objective 4: The target audiences for this project include: melon growers, packers, processors, and consumers. Produce growers and manufacturers of natural antimicrobials have been involved in this project by providing in-kind support to the project. Industry and consumers have been informed periodically on the results of this project through various forums such as seminars, oral and poster presentations, workshops, webinars, and demonstrations. Objective 5: HRD scholars and practitioners, General community such as consumers and retailers and other public stakeholders, Food safety scientist's community. Changes/Problems:NIL What opportunities for training and professional development has the project provided?Objective 1: Nutrition and quality team: Four Post-Doctoral Research Associates, four undergraduate student workers, seven graduate students, and one Research Assistant assisted with the project activities. Sensory team: part-time employees participated in the sensory evaluation study. Objective 2: A Postdoctoral fellow participated in data analysis of melon fruit transcriptome and metabolome. Two graduate students, Kai He and Nami Niyakan, have developed the RNA-seq data analysis pipeline and performed differential gene expression and temporal RNA-seq data analyses. Kai has graduated in Summer 2020 and joined Qualcomm. Objective 3: Arizona: Senior Farm Attendant has learned advanced growing and irrigating techniques each year of the project. North Carolina: The studies in NC have provided the opportunity to involve 4 recently graduated undergraduates, two undergraduates, two graduate students, and a Research Associate. Each of these individuals will be educated as to the development of melons, new cultivar development, evaluation of quality characteristics (i.e. Brix), and collection of yield and fruit size data. This experience may provide opportunities for new jobs. Texas-Uvalde: Two undergraduate student interns and one technician were trained/hired (partial funds) to assist with project activities. Texas-Weslaco: Three undergraduate student interns and two agricultural technicians were hired/trained to assist with project activities. California: One program technician and three technicians that work for the UC WSREC have increased their knowledge of melon quality evaluation. Objective 4: Food safety team: Training was provided to nine undergraduate student volunteers and a high school student in media making, preparation of dilution buffers, dilutions, plating, and enumeration techniques. In addition, three undergraduate students and a research scientist were trained in microbiological techniques and the rest were involved in attachment strength experiments, calculation of attachment strength of foodborne pathogens, and preparation of samples for scanning electron microscopy. Six of the undergraduate volunteers were also trained in oral and writing communication skills by participating in various outreach activities and hands-on demonstrations offered for high school students. A research technician was trained in preparing scientific manuscripts for publication in peer-reviewed scientific journals. In Dr. Cooper's lab, one graduate student and two undergraduate students were trained on the proper techniques for collecting microbiome samples, processing of the samples, DNA extraction microbiome sequence library preparation, and microbiome data analysis. Objective 5: During the reporting period, the research team has recruited 01 postdoctoral research associate as a new member. Under the guidance of the supervisor, her tasks primarily include: 1) working on developing a manuscript for submission to journals and conferences and 2) establishing the logic models for evaluation of the interdisciplinary melon project. How have the results been disseminated to communities of interest?Objective 1:Dr. Bhimu Patil and his team members presented research outcomes as invited/ plenary/keynote speaker at ACS National Meeting & Exposition, III Asian Horticultural Congress - AHC2020 and 2021 ASHS Annual Conference. Objective 2: Results were disseminated as two peer-reviewed publications and two oral presentations. Objective 3: North Carolina: Results were disseminated with an abstract and by an oral zoom presentation at the ACS meeting in April. Indiana: We produced a video highlighting watermelon and melon variety trials conducted at the Southwest Purdue Ag Center. The video was part of the 2021 Southwest Purdue Ag Center virtual field day, which was held on June 24, 2021. We also developed a video highlighting specialty melon production in Indiana, the video was available for the 2021 small farm education webinar series which was held in Aug. 2021. Texas-Weslaco and Uvalde: Results of field evaluations from 2019 and 2020 evaluations were presented at ACS Conference. Agriculture and Food Chemistry Division Symposium. Apr 2021. Objective 4: Food safety team members disseminated the project findings at several events. Students, research technician, and research specialists presented their food safety research on the melon project at the 2020 virtual Annual Food Safety Conference hosted by the University of Arizona Food Safety Consortium in Tucson, Arizona. One graduate student and two undergraduate students participated in the poster competition and two of them won the second and third place awards. This conference was attended by the local produce and food industry professionals, regulatory agency scientists, and academic scientists. The team was interviewed by a Reporter for a 60 min Daily Wildcat Podcast regarding her Interest, career, and research in food safety and during this interview, she cited the melon project and the main focus of the project. Food safety team members presented the project findings at the virtual Annual Meeting of the International Association for Food Protection (IAFP). Richard Park presented a seminar on the project findings from the cross-contamination due to environmental matrices and the prevalence work at the School of Animal and Comparative Biomedical Sciences Fall Seminar Series at the University of Arizona. Objective 5: The research team has the following activities for dissemination purposes. 1) In October 2020, the research team attended the Virtual Food Safety Conference, organized by the University of Arizona. The technical and communicative aspects of food safety science highlighted the interdisciplinary nature of the Melon Project and the importance of dissemination of the melon benefits for the public good. 2) In February 2021, the research team presented at the virtual Academy of Human Resource Development (AHRD) Research Conference in the Americas along with a manuscript in their proceedings that was entitled "Application of the Logic Models of Evaluation to Multidisciplinary Research - The Case of a Federally Funded Project". In the oral presentation, the application of logic models for evaluation in consumers' human health and wellbeing that was associated with adding melons to their diet was emphasized with the community of HRD scholars and practitioners. 3) Due to covid-19, Patil organized virtual symposium "Beyond Chemistry: Consumer Acceptance of Flavor Food Safety and Health Benefits of Fruits and Vegetables" This symposium was sponsored by the Division of Agricultural and Food Chemistry of the American Chemical Society. The symposium attracted 44 multidisciplinary researchers from different countries to present their novel cutting-edge research pertaining to consumer acceptance, food safety, health benefits, novel processing, and application of nanotechnology of various fruit and vegetable crops. The first two sessions focused on the research presentations from 22 collaborators related to the SCRI_CAPS melon project. Presentations included melon cultivation, production practices that addressed major issues and critical needs faced by US melon industries. Also, high-quality, flavorful, nutritious, and safe melon varieties using state-of-the-art breeding practices were also presented. The logic models are illustrative and easy to understand. The logic model illustration is expected to reach the consumer audience in a consumer-friendly manner. The scheduled stakeholder engagement conference was postponed to Fall 2021 due to travel restrictions. What do you plan to do during the next reporting period to accomplish the goals?Objective 1: We will continue to implement a new economics experiment with eye-tracking devices, analyze the data and prepare a new presentation of the new experiment in professional meetings. All phytochemicals including volatile profiles from melons collected from all locations will be analyzed. Objective 2:Using a large RNA-seq and metabolomic dataset generated with 2019 samples from multiple varieties and locations, we will identify transcripts whose expression levels are highly correlated with metabolites of interest. We will start with the univariate analyses based on Pearson correlation to identify genes that are significantly correlated with corresponding metabolites. Multivariate analysis that considers potential confounding factors, including varieties, locations, and other sample conditions (virus infection), will be further conducted to identify potential genetic markers for metabolites of interest, in particular the ones related to fruit characteristics directly affecting melon supply and market. Objective 3: Arizona: We will collect commercially grown melons and environmental samples from Yuma area growers during the Spring 2021 planting season and send them to the University of Arizona for food safety analyses. North Carolina: The plan is to conduct one final study which is currently ongoing in 2021 and to summarize the results on the NC Cooperative Extension cucurbit portal at NC State University. We will be shipping melons to TX for phytochemical and sensory analysis and AZ for microbial analysis. This information will be collated and analyzed in several refereed publications. Texas-Uvalde: Currently, we are conducting another cultivar trial in a generalized RCBD design. This experiment includes four replicated plots of two commercial varieties, 'Da Vinci' and 'Infinite Gold', and eight TAMU hybrids, 'TH1', 'TH5', 'TH6', 'TH9', 'TH10', 'TH13', 'TH14', and 'TH16'. We plan to take yield and quality data as described above. In addition to these measurements, a graduate student working in Dr.Bhimu Patil's lab is taking sequential samples of developing fruits to assess metabolic changes across the course of fruit maturation. Texas-Weslaco: Due to severe weather damage (Valentine's Day Freeze, 2021) and COVID-19 restrictions, field operations, plant growth and yield performance was impacted. Yield and quality data from all the previous trials will be pooled and analyzed to identify lines with consistently superior performance under local production conditions. Data from all trial sites will also be subjected to a combined analysis to identify lines that are suitable for each production region. Texas-Pecos: TAMU hybrids, "TH-1, TH-5, TH-6, TH-9, TH-13, TH-14" and two commercial varieties "6C and NUN" from the Pecos area will be collected and send to VFIC, College Station for sensory and phytochemical analysis. Objective 4: Provided the effects of the pandemic are reduced, we will begin to safely collect additional melon samples from the commercial fields in Arizona and California for the prevalence work and process them for the pathogens and indicator microorganisms in this next reporting period. In addition, the Ravishankar laboratory will continue to collect samples from the growing regions to conduct studies on natural antimicrobial interventions, the attachment strength of foodborne pathogens, the transfer of foodborne pathogens via soil and dust and will write up the results for publication and dissemination to numerous target audiences. We will also continue the outreach activities. We will finalize the commercial melon microbiome analysis and publish the results in the next few months. The Cooper laboratory will also generate additional microbiome sequencing libraries for the different melon cultivars and melon samples from commercial fields at the point of harvest for year 4 of the project, and incorporate the data with year 1-3 data for the microbiome analysis and finalize the microbiome data analysis for the project. In Dr. Vitha's lab, the plan is to complete the analysis of selected samples and compare the surface texture results with the attachment study. Objective 5: Complete the logic model drawing for other objectives of the Melon project.

Impacts
What was accomplished under these goals? Objective 1: 10 commercial varieties and 9 hybrids were grown in 5 locations across the USA and shipped to TX to conduct sensory and phytochemical studies. Consumer preferred flavor attributes such as melon identity, sweet, sour, bitter, overall sweet, musty/earthy, honey, green, overripe, and fermented were evaluated. Surface moisture, hardness, and juiciness were evaluated as texture attributes. Hybrids TH5, TH6, TH1, and TH12 had the highest melon identity and sweet flavors. The characteristic melon color imparting compound β-carotene was significantly higher in the selected melon hybrids and cultivars (TH-6, TH-13, TH-14, and TT-DV) grown in Texas compared to TH-5, F-39, and HT-IG. The hybrids TH-5 and TH-6 grown in Arizona (AZ) and Texas-Uvalde (TXU) recorded higher β-carotene contents. Total soluble solids (TSS, °Brix) measures the sweetness and helps in determining the ripeness of fruits. The hybrids TH-5, TH-6, TH-13 and TH-14 grown in different locations showed stability in TSS contents. The cultivar TT-DV and HT-IG grew in Texas had higher TSS content compared to the cultivar F-39. Several "characteristics impact flavor or aroma compounds" (CIFAC) in Cucumis melo were identified from a total of 154 volatile compounds profiles generated across the melon samples. The total esters which contribute highly to the aroma of ripe melons were the highest in STA (Syngenta-Tarasco) from AZ and lowest in STR (T Rex) from TXU. The individual melon esters viz; ethyl benzoate, ethyl butanoate, benzyl acetate, and ethyl hexanoate were high in TH9, TH10, TH13, and SSU (Sugar shot) respectively, from California (CA) while ethyl 2-methylbutanoate was high in SSU from AZ. Another major aroma volatile in melon, the total alcohols was highest in TH13 from CA and the lowest in TH6 from AZ whereas (Z)-6-nonen-1-ol was the highest in TH11 from Texas-Weslaco (TXW). The aldehydes were found to be highest in TH9 and TH10 from TXU and lowest in TH13 from CA among which (Z)-6-nonenal, (E)-2-nonenal and (E,Z)-2,6-nonadienal were highest in STR from TXW, SSW (Sweet spring) and STR from AZ respectively. Besides the flavor and aroma imparting compounds, limonene and benzaldehyde which possess antimicrobial properties were also found in melon samples with the highest being in TH10 fruit from CA and TXU respectively. Objective 2: Using RNA-seq data from TXW grown melon varieties F39 and TTDV, we conducted differential gene expression analysis and identified genes enriched in fruit samples. Differentially expressed genes (DEGs) were first identified by comparing fruit gene expression profiles against leaf profiles. A total of 4469 DEGs in fruits were identified. DEGs were further analyzed for their expression trajectory during the time course of fruit development. Aromatic volatile data for corresponding fruit samples were collected, and relations between gene expression and metabolite profiles were analyzed. A shift of VOC profiles in both varieties was observed. The early-fruit (immature) profiles were rich in C5-C8 lipid-derived VOCs and the late-fruit (mature-ripened) profiles were abundant in C9 lipid-derived VOCs such as apocarotenoids, and esters. Several key transcripts in volatile biosynthetic pathway enzymes, such as CCD1 and CCD4 for the synthesis of apocarotenoids, and ripening regulatory genes such as NOR, RIN, and TAG1 homologs were identified, and their expression profile in relation to the overall metabolic shift was analyzed. Objective 3: During the spring 2021 growing season, 13 hybrids from TAMU breeding program and 5 varieties were planted in 6 locations (TX-Weslaco, TX-Uvalde, TX-Pecos, CA, NC and AZ). These varieties were grown following the commercial standard protocols used for growing melons. The trials were arranged as a randomized complete block design with four blocks. Sprinkler irrigation was used during germination, and a drip line system was used after germination to provide direct moisture to the roots while using water more efficiently. Varieties grown at these locations were harvested and shipped to College Station, TX for sensory and phytochemical analysis and to Tucson, AZ for food safety evaluation. Objective 4: 10 commercial varieties and 9 hybrids melon rinds were inoculated with either Salmonella enterica or Listeria monocytogenes in soil and dust. The melon hybrids TH9 and TH5 harbored the highest percent of Salmonella in their rinds while the percent transfer of L. monocytogenes was the highest for SMA (Mamut) and TH1, respectively. The lowest percent transfer of Salmonella and L. monocytogenes was recorded on TH1 and TH10, respectively. Overall, hybrids had higher and lower percent transfer for both Salmonella and Listeria from dust and soil compared to varieties. The attachment strength of Listeria monocytogenes and Salmonella Newport on melon hybrids were evaluated. The average attachment strength of S. Newport and L. monocytogenes on the melon hybrids was as follows: TH1, TH5, TH6, TH9, TH10, TH11, TH12, TH13, TH14, SAC (Accolade), SSW, SSA (Saurio), SMA, STR, STA, SSU. The strongest average attachment strength for S. Newport was on TH9, and the lowest was on SMA. The strongest average attachment strength for L. monocytogenes was on TH12 and the lowest was on TH1. After inoculating with S. Newport or L. monocytogenes cultures, the melon rinds were immersed in a plant-based antimicrobial solution, olive extract or oregano oil to study the antimicrobial activity. Phosphate buffered saline (PBS) was used as a control. The survivors of S. Newport and L. monocytogenes were enumerated at days 0 and 3 and compared to the control. The plant-based antimicrobial treatments caused significant log reductions in populations of Salmonella and L. monocytogenes. In general, oregano oil had better antimicrobial activity than olive extract and the treatments were more effective on Salmonella than on L. monocytogenes. The microbiome libraries for over 600 melon samples were sequenced to understand the bacterial communities present on the surface of melons at the time of purchase. Additionally, microbiomes from over 200 melon and soil samples from commercial melon fields at the point of harvest in Yuma, AZ, Imperial Valley, CA, and Huron, CA were analyzed. Objective 5: Due to covid, most outreach activities were virtually conducted. The logic model was developed based on the input from collaborators. Experiential learning opportunities to 9 undergraduate students were provided in relation to analytical skills of melon health properties and food safety issues. The research team also attended a virtual conference on Food Safety organized by the University of Arizona.

Publications

• Type: Conference Papers and Presentations Status: Other Year Published: 2021 Citation: Dhanani, T. and Patil, B. S. (2021). Nondestructive methods to determine the maturity of specific fruits and vegetables. ACS National Meeting & Exposition, April 5-16, 2021, San Antonio, TX.
• Type: Conference Papers and Presentations Status: Other Year Published: 2021 Citation: Jifon, J, Djidonou D., Leskovar D, Crosby K., and Patil B. S. (2021) Environmental and Genotypic Variation of Fruit Yield and Quality Traits in Melons. ACS National Meeting & Exposition, April 5-16, 2021, San Antonio, TX.
• Type: Conference Papers and Presentations Status: Other Year Published: 2021 Citation: Koiwa, H. (2021) Transcriptome and metabolome changes in melon fruits, ACS National Meeting & Exposition, April 5-16, 2021, San Antonio, TX.
• Type: Conference Papers and Presentations Status: Other Year Published: 2021 Citation: Qian, X. (2021) RNA-sequencing analysis to study melon transcriptome, ACS National Meeting & Exposition, April 5-16, 2021, San Antonio, TX.
• Type: Conference Papers and Presentations Status: Other Year Published: 2021 Citation: Schultheis, J.R. and K. Starke. (2021). Melon cultigens and their adaptation in the southeastern United States when grown in North Carolina. ACS National Meeting & Exposition, April 5-16, 2021, San Antonio, TX. 1. 2021
• Type: Conference Papers and Presentations Status: Other Year Published: 2021 Citation: Van, H. T. M., F. Nafukho, and B.S. Patil (2021). Application of the Logic Models of Evaluation to Multidisciplinary Research - The Case of a Federally Funded Project. Academy of Human Resource Development Annual Research Conference in the Americas. Feb 2021.
• Type: Conference Papers and Presentations Status: Other Year Published: 2021 Citation: Van, H. T. M., F. Nafukho, and B.S. Patil (2021). Communicating the Benefits of Consuming Safer and Healthier Melons via a Logic Model. ACS National Meeting & Exposition, April 5-16, 2021, San Antonio, TX.
• Type: Conference Papers and Presentations Status: Other Year Published: 2021 Citation: Park, R. (2021). Why We Peel Melons: A Peek into the Prevalence of Foodborne Pathogens Among Field-Grown Melons in Arizona and Environmental Risk Factors for Cross-Contamination. ACS National Meeting & Exposition, April 5-16, 2021, San Antonio, TX.
• Type: Conference Papers and Presentations Status: Other Year Published: 2021 Citation: Ravishankar, S. (2021). Attachment Strength of Foodborne Pathogens and Efficacy of Plant-based Antimicrobials Against Salmonella enterica and Listeria monocytogenes on Melons Grown in Different Regions of the United States. ACS National Meeting & Exposition, April 5-16, 2021, San Antonio, TX.
• Type: Conference Papers and Presentations Status: Other Year Published: 2021 Citation: Vitha, S. (2021). Surface texture of honeydew & cantaloupe melons and its correlation with bacterial attachment. ACS National Meeting & Exposition, April 5-16, 2021, San Antonio, TX.
• Type: Conference Papers and Presentations Status: Other Year Published: 2020 Citation: Rossete, J., Rao, A., Porchas, M., Patil, B., Brierley, P. and Ravishankar, S. (2020). Determination of the Shelf-Life and Keeping Quality of Melons Grown in Different Regions with Respect to Background Microflora, Prevalence of Foodborne Pathogens and Variances in Netting Densities. Presentation at the Annual Food Safety Conference, Tucson, AZ, October 21, 2020.
• Type: Conference Papers and Presentations Status: Other Year Published: 2020 Citation: Zhu, L., Wei, Q., Brierley, P., Porchas, M., Patil, B. and Ravishankar S. (2020). Plant-based Antimicrobials Inactivate Salmonella enterica and Listeria monocytogenes on Melon Rinds. Presentation at the Annual Food Safety Conference, Tucson, AZ, October 21, 2020.
• Type: Conference Papers and Presentations Status: Other Year Published: 2020 Citation: Park, R., Rowlands, D., Porchas, M., Brierley, P., Patil, B. and Ravishankar, S.(2020). Understanding the Cross-contamination of Melons Via Environmental Matrices Under Field Conditions and Prevalence of Foodborne Pathogens. Presentation at the Annual Food Safety Conference, Tucson, AZ, October 21, 2020.
• Type: Conference Papers and Presentations Status: Other Year Published: 2020 Citation: Park, R. & Ravishankar, S. (2020). Why We Peel Melons: A Peek into the Prevalence of Foodborne Pathogens Among Field-Grown Melons in Arizona and Environmental Risk Factors for Cross-Contamination. Seminar presented at the ACBS Fall 2020 Seminar Series. Tucson, AZ, October 26, 2020.
• Type: Conference Papers and Presentations Status: Other Year Published: 2020 Citation: Zhu, L., Wei, Q., Brierley, P., Porchas, M., Patil, B. and Ravishankar S. (2020_. Plant-based Antimicrobials Inactivate Salmonella enterica and Listeria monocytogenes on Melon Rinds. Presentation at the Annual Meeting of the International Association for Food Protection, October 26-28, 2020.
• Type: Conference Papers and Presentations Status: Other Year Published: 2020 Citation: Park, R., Rowlands, D., Porchas, M., Brierley, P., Patil, B. and Ravishankar, S. (2020). Understanding the Cross-contamination of Melons Via Environmental Matrices Under Field Conditions and Prevalence of Foodborne Pathogens. Presentation at the Annual Meeting of the International Association for Food Protection, October 26-28, 2020.
• Type: Conference Papers and Presentations Status: Other Year Published: 2020 Citation: Ravishankar, S. (2020). Adoption of Emerging Technologies by the Food Industry Worldwide: Issues and Challenges under the symposium session titled A Global Perspective on New Generation of Food Processing/Preservation Techniques for Food Safety: Riding the Tides of Clean Labels. Annual Meeting of the International Association for Food Protection, October 28, 2020.
• Type: Conference Papers and Presentations Status: Other Year Published: 2020 Citation: Ravishankar, S. (2020). Current Trends in Food Safety and Food Production as part of the webinar series "Novel Strategies in Drug Development virtual presentation, Hindustan College of Arts & Sciences, Coimbatore, Tamil Nadu, India, on December 3, 2020.
• Type: Journal Articles Status: Published Year Published: 2021 Citation: Singh, J., Metrani, R., Jayaprakasha, G. K., Crosby, K., Ravishankar, S. and Patil, B. S. (2021). Improved sample preparation and optimized solvent extraction for quantitation of carotenoids. Plant Foods for Human Nutrition 76 (1), 60-67.
• Type: Journal Articles Status: Published Year Published: 2021 Citation: Ravindranath, V., Singh, J, Jayaprakasha, G. K., and Patil B. S. (2021). Optimization of Extraction Solvent and Fast Blue BB Assay for Comparative Analysis of Antioxidant Phenolics from Cucumis melo L. Plants 10 (7), 1379.
• Type: Journal Articles Status: Published Year Published: 2021 Citation: Tabara, M., Nagashima, Y., He, K., Qian, X., Crosby, K.M., Jifon, J., Jayaprakasha, G.K., Patil, B., Koiwa, H., Takahashi, H., and Fukuhara, T. (2021). Frequent asymptomatic infection with tobacco ringspot virus on melon fruit. Virus Res 293, 198266.
• Type: Conference Papers and Presentations Status: Other Year Published: 2021 Citation: Metrani, R., Singh, J., Crosby, K., and Patil, B. (2021). Aroma Volatile Profile of Cantaloupes (Cucumis melo L.) from different cultivars grown in five locations in the United States. ACS National Meeting & Exposition, April 5-16, 2021, San Antonio, TX.
• Type: Journal Articles Status: Published Year Published: 2021 Citation: Nagashima, Y., He, K., Singh, J., Metrani, R., Crosby, K.M., Jifon, J., Jayaprakasha, G.K., Patil, B., Qian, X., and Koiwa, H. (2021). Transition of aromatic volatile and transcriptome profiles during melon fruit ripening. Plant Science: an international journal of experimental plant biology 304, 110809.
• Type: Journal Articles Status: Published Year Published: 2021 Citation: Zhu, L., Wei, Q., Porchas, M., Brierley, P., Friedman, M., Crosby, K., Patil, B., and Ravishankar, S. (2020). Plant-Based Antimicrobials Inactivate Listeria monocytogenes and Salmonella enterica on Melons Grown in Different Regions of the United States. Food Microbiology (2021): 103876.
• Type: Journal Articles Status: Published Year Published: 2021 Citation: Perera, W.H., Shivanagoudra, S. R., P�rez, J. L., Kim, D. M., Sun, Y., Jayaprakasha, G. K., and Patil, B. S. (2021). Anti-Inflammatory, antidiabetic properties and in silico modeling of cucurbitane-type triterpene glycosides from fruits of an Indian cultivar of Momordica charantia L. Molecules 26 (4), 1038.
• Type: Conference Papers and Presentations Status: Other Year Published: 2020 Citation: Patil, B.S., Jayaprakasha, G.K., Acharya, P., Singh, J., (2020). The Nexus of Postharvest, food processing, nutrition, malnutrition and climate change. III Asian Horticultural Congress - AHC2020. Dec 15 to 17, 2021, Bangkok, Thailand.
• Type: Conference Papers and Presentations Status: Other Year Published: 2021 Citation: Singh J., Metrani, R., Crosby, K.M., Jifon, J., Ravishankar, S., Patil, B.S., (2021). Assessment of carotenoids and sugars contents in melon cultivars harvested from five different locations in the United States. ACS National Meeting & Exposition, April 5-16, 2021, San Antonio, TX.
• Type: Conference Papers and Presentations Status: Other Year Published: 2021 Citation: Singh J., Metrani, R., Crosby, K.M., Jifon, J., Ravishankar, S., Patil, B.S., (2021). Comparative Phytochemical Profiling of different Melon Hybrids Grown in Different Locations in the United States. 2021 ASHS Annual Conference, Aug 5 to 9, 2021, Denver, Colorado.
• Type: Journal Articles Status: Published Year Published: 2020 Citation: Wang, J., Jayaprakasha, G. K., and Patil, B. S. (2020). Untargeted Chemometrics Evaluation of the Effect of Juicing Technique on Phytochemical Profiles and Antioxidant Activities in Common Vegetables. ACS Food Science & Technology 1 (1), 77-87.
• Type: Journal Articles Status: Published Year Published: 2020 Citation: Kasote, D. M., Jayaprakasha, G. K., Ong, K., Crosby, K. M., and Patil B. S. (2020). Hormonal and metabolites responses in Fusarium wilt-susceptible and -resistant watermelon plants during plant-pathogen interactions. BMC plant biology 20 (1), 1-12.
• Type: Journal Articles Status: Published Year Published: 2021 Citation: Lee, J. H. J., Jayaprakasha, G. K., Enciso, J., Avila, C. A., Crosby, K. M., and Patil B. S. (2021). Production system influences tomato phenolics and indoleamines in a cultivar-specific manner. Food Research International 140, 110016
• Type: Journal Articles Status: Published Year Published: 2021 Citation: Lee, J. H. J., Kasote, D. M., Jayaprakasha, G. K., Avila, C. A., Crosby, K. M., and Patil B. S. (2021) Effect of production system and inhibitory potential of aroma volatiles on polyphenol oxidase and peroxidase activity in tomatoes. Journal of the Science of Food and Agriculture 101 (1), 307-314.
• Type: Journal Articles Status: Published Year Published: 2020 Citation: Lee, J. H. J., Jayaprakasha, G. K., Avila, C. A., Crosby, K. M., and Patil B. S. (2020). Effects of genotype and production system on quality of tomato fruits and in vitro bile acids binding capacity. Journal of Food Science 85 (11), 3806-3814.
• Type: Conference Papers and Presentations Status: Other Year Published: 2021 Citation: Patil, B.S., Singh J., Metrani, R., (2021). Optimized methods for rapid and sensitive analysis of volatiles and carotenoids from Muskmelons (Cucumis melo L.). 2021 ASHS Annual Conference, Aug 5 to 9, 2021, Denver, Colorado.
• Type: Conference Papers and Presentations Status: Other Year Published: 2021 Citation: Ravindranath, V., Metrani, R., Leskovar, D., Singh J., Jayaprakasha, G.K., Patil, B.S., (2021). Chemometric study of post-harvest storage effects on bioactive compounds in melon cultivars. ACS National Meeting & Exposition, April 5-16, 2021, San Antonio, TX.
• Type: Conference Papers and Presentations Status: Other Year Published: 2021 Citation: Acharya, P., Singh, J., Jayaprakasha, G.K., Patil, B.S., (2021). Nanoparticle-mediated seed priming: an effective technique to improve germination, growth, yield while maintaining the quality of watermelon. ACS National Meeting & Exposition, April 5-16, 2021, San Antonio, TX.
• Type: Conference Papers and Presentations Status: Other Year Published: 2021 Citation: Perez J.L., Singh, J., Patil, B.S., (2021). Bitter melon: Exploring the nutritional potential of an underutilized cucurbit. ACS National Meeting & Exposition, April 5-16, 2021, San Antonio, TX.
• Type: Conference Papers and Presentations Status: Other Year Published: 2021 Citation: Majithia, D., Metrani, R., Dhowlaghar, N., Jayaprakasha, G. K., Crosby, K., and Patil, B. S. (2021). Characterization of volatile profiles in 28 melon breeding lines using headspace solid-phase microextraction gas chromatography-mass spectrometry. ACS National Meeting & Exposition, April 5-16, 2021, San Antonio, TX.
• Type: Conference Papers and Presentations Status: Other Year Published: 2021 Citation: Metrani, R., Dhowlaghar, N., Jayaprakasha, G. K., Crosby, K., and Patil, B. S. (2021). Study of potential antimicrobials from melon (Cucumis melo L.) extracts. ACS National Meeting & Exposition, April 5-16, 2021, San Antonio, TX.

Progress 09/01/19 to 08/31/20

Outputs
Target Audience:Objective 1:For the sensory evaluation, data were presented to sensory professionals across the globe on how to evaluate melons and relate sensory descriptive attributes to consumer liking and preference. Target audiences include scientific colleagues and students through conference talks, university seminars, and lectures. Finally, the outcome of the research will be helpful for producers and seed companies. The National Center of Excellence for Melons is physically located at the Vegetable and Fruit Improvement Center, Texas A&M University, College Station. This Center is visited by several target audienceon a routine basis. The target audiences include master gardeners, high school and potentials students, legislatures and their aids, the scientific community, industry professionals, regulatory scientists, and academicians. Similarly, UA Food Safety lab is visited by similar audiences. Investigators provide the tour and present the results of the melon project along with other Projects. Objective 2:Commercial growers and one seed company (Gowan seeds) Objective 3:Consumers, growers, retailers, seed/agrochemical companies, the scientific community, home gardeners, and state (Texas Department of Agriculture) representatives. Georgia- Agricultural professionals and those in the vegetable industry that have read the popular press article have been reached. Growers have not yet been given the results as the trial season has just been completed. North Carolina- The main target audience for these field studies is the agricultural industry which includes seed companies and producers. These audiences can easily access the results remotely through the internet. Objective 4:The target audiences for this project include melon growers, packers, processors, and consumers. Produce growers and manufacturers of natural antimicrobials have been involved in this project by providing in-kind support to the project. They have been informed periodically on the results of this project through various forums such as seminars, oral and poster presentations, workshops, webinars, and demonstrations. Changes/Problems:Due to resignation of co-investigator at University of Florida, the project at Flordia was dropped. The same resources are used to expand the project in Georgia, North Carolina and Indiana instead of original plan from 2 to 4 years. Due to Covid-19, North Carolina and Georgia could not conduct field studies in 2020. Both states have agreed to conduct field studies in the coming years. What opportunities for training and professional development has the project provided?Objective 1: Four undergraduate student workers, six graduate students, and six part-time sensory employees participated in the study for sensory evaluation. One PhD students trained in the auction mechanism and experimental design for consumer panel. A postdoctoral researcher, two graduate, one research assistant, and two undergraduate students received training in phytochemicals, volatiles, postharvest and related quality attributes. Postdoctoral research Associate trained undergraduate students. Objective 2: One postdoctoral, two graduate student, 1 visiting PhD student, one research assistant, and 2 undergraduate students were trained in all aspects of melon breeding, selection and quality analyses and RNA sequence. Objective 3: Arizona - Our Senior Farm Attendant has learned advanced growing and irrigating techniques each year of the project. This year's concerns of fly larva destroying the seedlings required him to consult with University of Arizona research entomologists to devise a growing plan to protect the crop. The vine decline which plagued the Arizona melons last year also provided a hands-on professional development opportunity for our Senior Farm Attendant as he worked closely with the Yuma Ag Center Farm Manager and Superintendent and University of Arizona research scientists to mitigate damage to the melons and select a new field site for this year. California - One program technician and three technicians that work for the UC WSREC have increased their knowledge of melon quality evaluation. Georgia- A student participated in the project as part of a young scholars program at The University of Georgia. Information generated from this project will also be used for grower meetings during the winter of 2020. North Carolina- The cultigen evaluation study conducted in 2019 in North Carolina involved one research associate, four undergraduate students and one post baccalauareate employee that were involved in the vegetable production/cultural management program. In addition, four full time employees at the Central Crops Research Station cared for the field study throughout the growing season and assisted with harvest. Indiana- A technician and about 5 undergraduate students from Horticulture department in Vincenne University have worked with this project. They are trained in melon production. Texas -Weslaco: Two undergraduate student interns and two ag technicians were hired/trained to assist with project activities. Texas -Uvalde: Two undergraduate student interns and one technician were trained/hired (partial funds) to assist with project activities. Objective 4: Food Safety team' lab has provided the opportunity for one research specialist, four research technicians, one graduate student, and nine undergraduate students to learn numerous techniques and skills related to food safety research, including but not limited to sampling of melons and environmental samples for foodborne pathogens and indicator microorganisms, natural antimicrobial interventions for melon post-harvest treatment, measurement of bacterial attachment strength, assessment of transfer rate of pathogens, antimicrobial treatments, biophotonic imaging and sample preparation for scanning electron microscopy. Cooper's lab has provided the opportunity for a new research technician to learn skills related to food safety research including culturing pathogens from different types of samples, collecting and processing microbiome samples, and sample collection of various types of samples directly from the field. The project has also provided the opportunity for two graduate students and seven undergraduate students the opportunity to learn and be trained on the proper techniques for collecting various types of samples directly from melon fields, processing of the samples, culturing the samples for pathogens, DNA extraction, microbiome sequence library preparation, and microbiome data analysis and also interacting with different individuals in the melon industry. How have the results been disseminated to communities of interest?Objective 1: Miller, presented poster in August at the Pangborn International Symposium for Sensory Evaluation. Patil has presented research outcomes as invited/plenary/keynote speaker in several national and international conferences organized by Global Conference on Obesity, National Conference on Arid Fruits, and 5th IUPHAR Word Conference on the Pharmacology of Natural Products and 51st Annual Conference of Indian Pharmacological Society Nutrition and Health Disorders. Objective 2: We planted field trials with two commercial growers in Texas, J&D Produce and Mandujano Farms. We received positive feedback from J&D and are analyzing the data while the Mandujano trial is not mature yet. Objective 3: Arizona- Arizona's Yuma Center of Excellence for Desert Agriculture shared the melon project's status at its March 2020 donor meeting. Arizona plans to host a stakeholder conference in 2022. Georgia- The results have recently been compiled and will be disseminated to growers until winter meetings. Two years of trials complete results will be disseminated in the form of extension publications. One popular press article has been published (http://www.caes.uga.edu/news-events/news/story.html?storyid=7645&story=Cantaloupes). This story has been picked up by several online outlets that reach vegetable growers. North Carolina- The results have been disseminated via an electronic publication that can be accessed on the internet 24 hours a day 7 days a week at the following link. https://cucurbits.ces.ncsu.edu/growing-cucurbits/variety-trials/2019-north-carolina-orange-flesh-and-specialty-melon-cultigen-evaluations/Indiana- The trial results were shared with melon growers and seed company representatives at the Southwest Indiana Melon and Vegetable Growers Winter Meeting on Nov. 14, 2019. The meeting had about 50 attendances. A written report was published on Midwest Vegetable Trial Report. https://docs.lib.purdue.edu/cgi/viewcontent.cgi?article=1015&context=mwvtr. Texas- Weslaco: An open field visitation day was held prior to harvests and was attended by local growers and seed company representatives. Results of field evaluations from 2019 were presented at a local grower meeting in Weslaco, TX. Texas -Uvalde: An open field day was held during the spring field day in 2019. About 80 participants toured the melon field plots prior to harvest. The field day for spring-summer 2020 was cancelled due to COVID19. Objective 4: Students, research technicians, research specialist and research scientist in Food Safety lab presented their food safety research on the melons project at the 2019 Annual Food Safety Conference hosted by the University of Arizona Food Safety Consortium in Tucson, Arizona. This conference was attended by the local food industry professionals, regulatory agency scientists, and academic scientists. Dr. Ravishankar and her team members, including research scientist, research specialist, research technicians, and students, had the opportunity to participate in the food safety poster session at the 2020 Southwest Agricultural Summit held in Yuma, Arizona to provide information to the stakeholders on melons and other produce safety issues and interventions. Demonstrations on the use of plant antimicrobials and other aspects of melons food safety research were conducted for the produce industry professionals, University of Arizona alumni/donors and local high school students throughout the year. Objective 5. Results of all the objectives were presented to the Advisory Board (Principal Stakeholders and Scientific Managing Team) by each team lead leaders. Priority areas and potential impacts were discussed at length. What do you plan to do during the next reporting period to accomplish the goals?Objective 1: Varieties across locations that were selected based on transidisciplinary experts including sensory, consumer, breeder, crop management and breeders, will be grown as defined for the last 3 years. These varieties across different locations will be evaluated for food safey, quality and health promoting properties. Phytochemicals such as carotenoids, vitamin C, phenolics, amino acids and aroma volatile compounds from two commercial varieties (TTDV and F39), nine new hybrids and seven hybrid melon lines from Syngenta seeds grown in all locations will be analyzed. Furthermore, antioxidant activity and polyphenol oxidase using different in-vitro assays from these varieties will be determined. Due to COVID 19, several partner states could not grow melons and it is expected that the coming year, we will grow melons in all partner states. This plan is still fluid due to uncertainties of resurgence of COVID 19. Each team leads and the respective members will prepare manuscripts and present their data in professional meetings and industry groups. Objective 2: Continue data analysis of RNA seq samples to obtain maturation associated transcripts in F39 and TTDV. Analyze RNA-seq data from melon samples grown at different locations to conduct association analysis. Conduct controlled crossing in the greenhouse to generate additional hybrids based on the combining ability estimates from the field trials and the fruit quality attributes of the inbred lines evaluated, focusing on firmness and brix as the key attributes while eliminating any disease susceptible parents. Plant new F1 hybrids from the GH crossing, breeding lines, and some F2 populations for field selection at Weslaco, Uvalde and Pecos. Prepare release documentation for a new hybrid and its inbred parent lines. Prepare a publication on melon fruit quality traits heritability estimates. Objective 3: Arizona- A second group of samples, consisting of only commercial varieties, will be collected during the fall 2020 growing season and shipped to food safety at The University of Arizona for testing. Corresponding environmental samples will also be collected and shipped along with the commercial varieties. YCEDA will conduct variety field trial using Arizona commercial methods during the 2021 spring growing season. A Yuma conference will be planned and held to disseminate results after the Texas conference. California- The melon field comparison at the site in western Fresno County will be monitored for seasonal development, and yield and quality of the melons will be determined. At maturity, representative melons will be selected, packed and shipped to laboratories in Texas and Arizona for additional assessments. Information will be made to clientele at an extension meeting to be held at the Research and Extension Center in Fresno County or on a Zoom extension meeting to be held in late 2020. Information on the three years of comparisons will be shared with clientele through a report to be made available through the University of California Ag and Natural Resources web site. North Carolina- Duo to COVID-19, we were not able to conduct the study. We plan to resume field testing in 2021. Indiana- Fourteen commercial cantaloupe cultivars and advanced breeding lines will be evaluated in the 2020 season at Southwest Purdue Agriculture Center in Vincennes, IN. The trial results will be shared with growers at meetings and through published trial report. Texas- Uvalde: We plan to sample multiple soil cores from the interspaces in each plot to measure root length density and specific root length. We anticipate taking these measurements twice during the remaining stages of plant development and maturation along with additional physiological measurements. We plan to replicate the methods used last year to measure yield and quality. Briefly, harvested fruits will be graded according to U.S. commercial trade standards then counted and weighed for each grade. Total and marketable yields will be estimated from this data. Randomly selected fruits will be sampled for physical (individual fruit weight, length, and diameter) and tissue quality traits (firmness, color, and soluble solids content) and other quality, safey, and health properties. Weslaco: Fruit yield and quality data from all the previous trials will be pooled and analyzed to identify lines with consistent superior performance under local production conditions. Data from all trial sites will also be subjected to a combined analysis to identify lines that are suitable for each production region. Objective 4: Due to the COVID-19 pandemic we will not be able to safely collect additional melon samples from the fields in Arizona and California and process them for the pathogens in this next reporting period, but will resume once it is safe to conduct the research. However, we will continue to collect samples from the growing regions to conduct studies on natural antimicrobial interventions, attachment strength of foodborne pathogens, the transfer of foodborne pathogens via soil and dust and will write up the results for publication and dissemination to numerous target audiences. We will also continue the outreach activities. Also, we will finalize the commercial melon microbiome analysis and publish the results. The Cooper laboratory will also finalize the sequencing libraries for the different melon cultivars and melon samples from commercial fields at the point of harvest, get the libraries sequenced, conduct the data analysis for both projects and publish the results. Objective 5 Due to Covid 19, scheduled stakeholders-engagement conference is postponed to June-July 2021. Develop bilingual short course material for Master Gardeners. Continue to provide experience learning opportunities to undergraduate students both in the lab and produce industry.

Impacts
What was accomplished under these goals? Objective 1: and 6 hybrids were grown in 7 locations and shipped to TX to conduct flavor, sensory and quality including health promoting properties as well as to AZ to conduct fruit safety analysis. Flavor attributes evaluated were melon identity, sweet, sour, bitter, overall sweet, musty/earthy, honey, green, overripe, and fermented. Surface moisture, hardness and juiciness were evaluated as texture attributes. Consumers acceptance for four varieties from Weslaco rated these melons for purchase price, appearance and aroma liking, and cut melon appearance, aroma, overall, flavor and texture liking. Varieties that had the highest melon identity and sweet flavors were liked by consumers to a greater degree. We found F39 attained the highest price in both appearance and taste bidding rounds, while TTDV received the highest bid in both bidding rounds. The GABA (?-amino bytyric acid) and vitamin C content in melons range from 572.11 to 2582.93, respectively. A total of 189 volatile compounds were identified and quantified. Esters that impart fruity aroma where positively correlated with TTDV from all the locations. Aldehydes which imparts melon green aroma were high in TH6 and TH3 from Arizona as well as honeydew varieties HD150, HD252 and OC164. On the other hand, cucumber like odor was positively correlated to HD150, HD252 and OC164. The compounds, which imparts fruity and sulphurous aroma were positively correlated with F39 and TTDV. The selected antimicrobial compounds were significantly higher in cantaloupes compared to honeydews. A total of 29 breeding lines were grown at Weslaco and 113 volatile compounds were identified which are responsible for the flavor and aroma profiles of musk-melons. The highest levels of average volatile concentrations were found in BL30 and the lowest in BL22. Objective 2: RNA-seq for 92 melons and 3 leaf samples as references was conducted and fruit maturation time course samples were processed as a pilot-scale gene expression analysis. Samples were collected as triplicates, using two varieties (F39 and TTDV), harvested as early, (day 25), middle (day 30), late (day 35) and mature (day 40) stages. We identified approximately 4000 genes differentially (>3 fold) expressed in fruits relative to leaves in each variety. The genes identified are being grouped for their expression patterns during the maturation, and their functions studied using gene ontology analysis and individual annotations. F39 and TTDV have distinct kinetics of transcriptome changes. While F39 progressively shifts from immature to mature profiles over the time, TTDV kept the immature profiles until mid-stage then quickly switch to the mature profiles. At the individual sequence levels, polymorphisms in several SNP that affect carotenoid levels and viral disease resistance was identified. Field trials at Weslaco and Uvalde of 34 new hybrids and 28 elite inbred lines was completed. Fruit quality traits: size, color, brix, flesh firmness, cavity size, rind netting, peduncle scar size, shape, and yield was collected. Four hybrids with extremely firm flesh, >80 N, based on digital penetrometer readings were identified. Additionally, 14 of the hybrids had brix of 11. Fruit size ranged from 6 to 15 and yields ranged from 1 to 2.2 boxes per 10 ft plot. Vine health and resistance to powdery mildew, downy mildew and Monosporascus root rot and vine decline were recorded in the field plots. Resistance to all three diseases was identified in several hybrids and inbred lines. In the greenhouse, production of additional hybrid seed for the best 6 hybrids and some of the elite parent lines was conducted by controlled pollinations. Additionally, 3 rounds of controlled inoculations of an F2 population and its parents was conducted with Monosporascus cannonballus to rate resistance in the roots. DNA was extracted and sequenced to conduct SNP marker development to construct a genetic linkage map and identify SNP's linked to the resistance QTL. Resistant F2 progeny were saved to backcross to the desirable muskmelon parent to introgress the resistance QTL into a high quality melon background. Objective 3: During the spring 2020 growing season, eight hybrids from TAMU breeding program and seven hybrid lines from Syngenta were planted in five states. These varieties were grown following commercial standard protocols used for growing cantaloupe. The trial was arranged as a randomized complete block design with four blocks. Sprinkler irrigation was used during germination, and a drip line system was used after germination to provide direct moisture to the roots while using water more efficiently. Varieties grown at these locations were harvested and shipped to College Station and to Tuscan for laboratory analysis and food safety evaluation. Objective 4: Melon rinds inoculated with S. Newport or L. monocytogenes culture were tested for their survivor at Day 0 and 3. The plant-based antimicrobials (5% olive extract or 0.5% oregano oil antimicrobial solution) reduced S. Newport and L. monocytogenes population on all the rind samples. Compared to the control wash (PBS), the plant-based antimicrobial treatments caused significant log reductions in populations of Salmonella and L. monocytogenes. In most cases, the plant-based antimicrobial treatments reduced the pathogen population to below the detection limit at day 3. In general, oregano oil had better antimicrobial activity than olive extract. The antimicrobial treatments exhibited better reductions on honeydews than on cantaloupes. A total of 339 cantaloupes harvested from four fields in Arizona and 300 melons from California, along with soil, rhizosphere, air, and water environmental samples were tested for the presence of Listeria monocytogenes and Salmonella. Neither Listeria monocytogenes nor Salmonella were found in either the melon composites or environmental samples from Arizona field, while 3/32 air samples and 1/285 melon samples from California were positive for Salmonella. Melon rinds were sprayed with dust inoculated with either Salmonella or L. monocytogenes. Percent transfer of dust and soil for Salmonella was the highest for HTIG-AZ and TH6-TXU respectively. Percent transfer of L. monocytogenes was highest of dust and soil for F39-AZ and TH7-AZ respectively. In general, hybrids had higher percent transfer for both Salmonella and Listeria from soil than the varieties. Hybrids also had the lowest percent transfer from dust for both Salmonella and L. monocytogenes as compared to the varieties. A comparison between cantaloupes and honeydews showed that honeydews had the lowest percent transfer for both pathogens from soil. In case of dust, honeydews and cantaloupes showed the highest % transfer for Salmonella and L. monocytogenes, respectively. Melon rind was inoculated with foodborne pathogens to calculate the attachment strength. In general, the attachment strength of Salmonella was higher on cantaloupes than honeydews, while L. monocytogenes showed higher attachment strength on honeydews than cantaloupes. L. monocytogenes showed higher attachment strength on hybrids than cantaloupe varieties, while Salmonella showed slightly higher attachment strength on the cantaloupe varieties in comparison to the hybrid. Selected samples are being analyzed for surface roughness via scanning electron microscopy. Microbiome studies: The microbiome libraries for over 400 commercial melons were sequenced to understand those bacterial communities that are present on the different melon surfaces at the time of purchase. We have also prepared the sequence libraries for most of different melon cultivars grown in the different states involved in this project.

Publications

• Type: Journal Articles Status: Published Year Published: 2020 Citation: Singh, J., Metrani, R., Jayaprakasha, G. K., Crosby, K., Ravishankar, S. and Patil, B. S.2020. Multivariate Analysis of Amino Acids and Health Beneficial Properties of Cantaloupe Varieties Grown in Six Locations in the United States. Plants 9:1058.
• Type: Journal Articles Status: Published Year Published: 2020 Citation: Vitha, S. and Ravishankar, S. (2020). Surface texture analysis of melon rind using 3D reconstruction from SEM stereo images. Texas Journal of Microscopy. 51:24.
• Type: Journal Articles Status: Published Year Published: 2020 Citation: Metrani, R., Singh, J., Acharya P., Jayaprakasha, G. K. and Patil, B. S. Comparative metabolomics profiling of polyphenols, nutrients and antioxidant activities of two red onion (Allium cepa L.) cultivars. Plants 2020, 9, 1077.
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Jayaprakasha, G. K., Chidambara Murthy, K. N., Pellati, F. & Patil, B. S. BetaSweet carrot extracts have antioxidant activity and in vitro antiproliferative effects against breast cancer cells. Journal of Functional Foods 62: 103552.
• Type: Journal Articles Status: Published Year Published: 2020 Citation: Acharya P., Jayaprakasha, G. K., Crosby, K. M., Jifon, J. L. & Patil, B. S. Nanoparticle-Mediated Seed Priming Improves Germination, Growth, Yield, and Quality of Watermelons (Citrullus lanatus) at multi-locations in Texas. Scientific Reports (Nature Publisher Group) 10:5037.
• Type: Journal Articles Status: Published Year Published: 2020 Citation: Wang, J., Jayaprakasha, G. K. & Patil, B. S. UPLC-QTOF-MS fingerprinting combined with chemometrics to assess the solvent extraction efficiency, phytochemical variation, and antioxidant activities of Beta vulgaris L. Journal of Food and Drug Analysis 28:217-230.
• Type: Journal Articles Status: Published Year Published: 2018 Citation: Cordery, A., Rao, A. P., & Ravishankar, S. Antimicrobial Activities of Essential Oils, Plant Extracts and their Applications in Foods- A Review. Journal of Agriculture and Environmental Sciences, 7, 7689.
• Type: Book Chapters Status: Published Year Published: 2018 Citation: Ravishankar, S. Plant-Based Antimicrobials for Clean and Green Approaches to Food Safety. Natural and Bio-Based Antimicrobials for Food Applications, American Chemical Society, 2018, 4561.
• Type: Book Chapters Status: Published Year Published: 2019 Citation: Rao, A. P., & Ravishankar, S. Alternatives to Pest and Disease Control in Preharvest, and Washing and Processing in Postharvest Levels for Organic Produce." Safety and Practice for Organic Food, Academic Press, 2019, 213-226.
• Type: Book Chapters Status: Published Year Published: 2019 Citation: Taylor, T. M., Ravishankar, S., Bhargava, K., & Juneja, V. K. Chemical Preservatives and Natural Food Antimicrobials, Food Microbiology: Fundamentals and Frontiers, 2019, 705-731.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2019 Citation: Patil, B.S. Jayprakasha, G.K., and Singh, J. Can we reduce Obesity burden by consuming selected fruits and vegetables? Global Conference on Obesity, Nutrition and Health Disorders, October 7-8, 2019. Dubai, UAE.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2019 Citation: Patil, B.S. Jayprakasha, G.K., Achyara, P. and Singh, J. 2019. Arid Zone Fruits: A New Resource to Improve Human Health and Food Security in the Era of Climate Change. National Conference on Arid Fruits-A way forward for sustainable production and nutritional security. University of Agricultural Sciences, Raichur, India. November 28-30, 2019.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2019 Citation: Patil, B.S. Natural products from underutilized crops Isolation, identification, and examination of their role in mitigating cardiometabolic diseases. 5th IUPHAR Word Conference on the Pharmacology of Natural Products and 51st Annual Conference of Indian Pharmacological Society, December 5-6, 2019, Hyderabad, India.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2020 Citation: Metrani, R., Singh, J., Patil, B.S., Jayaprakasha, G.K. Qualitative and quantitative assessment of flavor from cucumis melo using Gas Chromatography electron and chemical ionization mass spectrometry. 68th ASMS Conference on Mass Spectrometry and Allied Topics, May-31-June 04, 2020, Houston, TX.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2020 Citation: Singh, J., Adhikari J., Jayaprakasha, G. K., Patil, B.S. Rapid UHPLC Identification of Anthocyanins from Different Varieties of Berries using High Resolution-QTOF-Mass spectrometry. 68th ASMS Conference on Mass Spectrometry and Allied Topics, May-31-June 04, 2020, Houston, TX.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2020 Citation: Ravindranath, V., Metrani, R., Singh, J., Patil, B.S., Jayaprakasha, G.K. Post-harvest storage influences volatile aroma profiles of melon varieties identified by HS-SPME-GC-MS. 68th ASMS Conference on Mass Spectrometry and Allied Topics, May-31-June 04, 2020, Houston, TX.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2019 Citation: Singh, J., Jayaprakasha, G. K., Crosby, K., Jifon, J., Brierley, P., Ravishankar, S., Leskovar, D., Turini, T., Schultheis, J., Coolong, T., Guan, W., Patil, B.S. Metabolic profiling of amino acids in cantaloupes harvested from different growing locations. Quantum chemistry and computational methods for compound identification, Metabolomics association of North America, summer symposium, September 04, 2019, UC Davis, CA.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2019 Citation: Metrani, R., Jayaprakasha, G. K., Crosby, K., Jifon, J., Brierley, P., Ravishankar, S., Leskovar, D., Turini, T., Schultheis, J., Coolong, T., Guan, W., Patil, B.S. A metabolomics approach to identify the changes in aroma volatiles in climacteric and non-climacteric melons grown seven locations. Quantum chemistry and computational methods for compound identification, Metabolomics association of North America, summer symposium, September 04, 2019, UC Davis, CA.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2019 Citation: Mkuu, R., Palma, M., Nafukho, F. M., & Patil, B. (2019). Consumers and industry representatives perceptions of quality, safety and attributes of melons for improved health outcomes. Academy of Human Resource Development International Research Conference, February 13-16, 2019, Louisville, KY.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2019 Citation: da Silva, A.L.B., Coolong T.W., Crosby., and Patil, B.S. Cantaloupe and Honeydew Yield and Quality Performance in Georgia. 116th annual ASHS Conference, July 21-25, 2019, Las Vegas, Nevada.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2020 Citation: Patil, B., Nafukho, F. M., & Van, H. T. M. (2020,). Application of the logic model of evaluation to food safety multidisciplinary research The case of the melon project. Association for International Agriculture and Rural Development 56th Annual Conference (AIARD), May 31-June 2, 2020, Washington, DC.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2019 Citation: Park, R., Rao A., Porchas, M., Patil, B., Brierley, P. and Ravishankar, S. Understanding the Cross-Contamination of Melons via Environmental Matrices Simulating Field Conditions. International Association for Food Protection 2019 Annual Meeting, July 22-24, 2019, Louisville, KY.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2019 Citation: Wei, Q., Rowlands, D., Torres, M., Porchas, M., Fang, T., Patil, B., Brierley, P. and Ravishankar, S. Attachment Strength of Foodborne Pathogens on Different Melon Varieties from Various Regions in the United States. International Association for Food Protection 2019 Annual Meeting, July 22-24, 2019, Louisville, KY.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2019 Citation: Rao, A., Park, R. Porchas, M., Patil, B., Brierley, P. and Ravishankar, S. Microbial Quality During Storage, Prevalence of Foodborne Pathogens and Salmonella Colonization Based on Variances in Netting Densities of Melons Grown in Different Regions of the United States. International Association for Food Protection 2019 Annual Meeting, July 22-24, 2019, Louisville, KY.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2019 Citation: Park, R., Rao A., Porchas, M., Patil, B., Brierley, P. and Ravishankar, S. Understanding the Cross-Contamination of Melons via Environmental Matrices Simulating Field Conditions. Annual Food Safety Conference, October 17, 2019, Tucson, AZ.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2019 Citation: Wei, Q., Rowlands, D., Torres, M., Porchas, M., Fang, T., Patil, B., Brierley, P. and Ravishankar, S. Attachment Strength of Foodborne Pathogens on Different Melon Varieties from Various Regions in the United States. Annual Food Safety Conference, October 17, 2019, Tucson, AZ.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2019 Citation: Rao, A., Park, R. Porchas, M., Patil, B., Brierley, P. and Ravishankar, S. Microbial Quality During Storage, Prevalence of Foodborne Pathogens and Salmonella Colonization Based on Variances in Netting Densities of Melons Grown in Different Regions of the United States. Annual Food Safety Conference, October 17, 2019, Tucson, AZ.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2020 Citation: Vitha, S. and Ravishankar, S. Surface Texture Analysis of Melon Rind Using 3D Reconstruction From SEM Stereo Image. Texas Society for Microscopy Conference, February 20-22, 2020, College Station, TX.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2020 Citation: Park, R., Rao A., Porchas, M., Patil, B., Brierley, P. and Ravishankar, S. Understanding the Cross-Contamination of Melons via Environmental Matrices Simulating Field Conditions. Southwest Ag Summit, February 25-27, 2020, Yuma, AZ.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2020 Citation: Wei, Q., Rowlands, D., Torres, M., Porchas, M., Fang, T., Patil, B., Brierley, P. and Ravishankar, S. Attachment Strength of Foodborne Pathogens on Different Melon Varieties from Various Regions in the United States. Southwest Ag Summit, February 25-27, 2020, Yuma, AZ.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2020 Citation: Rao, A., Park, R. Porchas, M., Patil, B., Brierley, P. and Ravishankar, S. Microbial Quality During Storage, Prevalence of Foodborne Pathogens and Salmonella Colonization Based on Variances in Netting Densities of Melons Grown in Different Regions of the United States. Southwest Ag Summit, February 25-27, 2020, Yuma, AZ.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2019 Citation: Rao, A., & Ravishankar, S. Cleaner and Greener Alternatives for Safe Food- Its Essential! Maricopa Environmental Health Services Department Training for Health Inspectors, April 2019, Phoenix, AZ.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2019 Citation: Rao, A., & Ravishankar, S. Cleaner and Greener Alternatives for Safe Food- Its Essential! USDA-Ag Discovery Camp students at the University of Arizona, June 2019.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2019 Citation: Ravishankar, S. (2019, June). Cleaner and Greener Alternatives for Safe Food- Its Essential! Sathyabama University, June 2019, Chennai, India.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2020 Citation: Park, R. Research Opportunities: Food Safety Lab- Creating a Safer Farm to Fork Journey. Faculty Data Blitz at the University of Arizona, February 28, 2020, Tucson, AZ.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2020 Citation: Ravishankar, S. Emerging Foodborne Microbial Threats: Challenges & Control Strategies. Invited International Webinar Zoom Talk Sponsored by Sathyabama Institute of Science & Technology, Indian Association of Applied Microbiologists, and California University of Science & Medicine- Webinar Series Novel Microbes & Newer Threats, May 2020.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2018 Citation: Fix, A., Lau, J., Crosby, K., Patil, B., Leskovar, D., Rooney, W., and Smith, W. Mission Melon: Improving Qualitative traits in Cucumis melo using Phenomics. Cucurbitaceae 2018. November 12-15, 2018, Davis, CA.

Progress 09/01/18 to 08/31/19

Outputs
Target Audience:Target audiences include scientific colleagues and students through conference talks, university seminars, and lectures. Finally, the outcome of the research will be helpful for producers and seed companies. The National Center of Excellence for Melons is physically located at the Vegetable and Fruit Improvement Center, Texas A&M University, College Station. This Center is visited by several target audience on routine basis. The target audiences include master gardeners, high school and potentials students, legislatures and their aids, the scientific community, industry professionals, regulatory scientists, and academicians. Similarly, UA Food Safety lab is visited by similar audiences. Investigators provide the tour and present the results of melon project along with other Projects. Arizona - Industry professionals/growers in the Imperial Valley, CA area and Arizona. Georgia - Agricultural professionals and those in the vegetable industry that have read the popular press article have been reached.Growers have not yet been given the results as the trial season has just been completed. Indiana - Indiana melon growers were aware of the project and expressed interest in learning project results. Texas (Uvalde)- consumers, growers, retailers, seed/agrochemical companies, scientific community Texas (Weslaco): consumers, growers, retailers, seed/agrochemical companies, scientific community, home gardeners, and state (Texas Department of Agriculture) representatives. California- consumers, growers, retailers, seed/agrochemical companies, scientific community North Carolina- consumers, growers, retailers, seed/agrochemical companies, scientific community Changes/Problems:There is no majpor change. One of the investigators from Florida is no longer involved in the project. We have decided to additional year with Georgia and North Carolina. What opportunities for training and professional development has the project provided? Objective 1.A postdoctoral researcher (Singh), three graduate students (Ravi, Majithia and Biradar), Research Assistant (Metrani), three undergraduate students received training in Phytochemcials, volatiles, postharvest and related quality attributes. Objective 2.Four graduate students (Fix, Marquez, Reyes, and He) were trained in all aspects of melon breeding, selection and quality analyses and RNA seq. Also, two technicians (Jones and Benitez) were trained in melon breeding, cultivation and data collection. Objective 3. Arizona - Martin Porchas was hired as a Farm Attendant to oversee the field trial in Yuma. One Technical team member was employed at 0.25 FTE for the field trial, which was performed by Martin Porchas, coordinated by Sonnet Nelson and overseen by Paul Brierley. Martin's professional development included additional experience and knowledge gained from experiment station personnel, making him more proficient at melon field trials. Objective 3. Arizona - Martin Porchas was hired as a Farm Attendant to oversee the field trial in Yuma. One Technical team member was employed at 0.25 FTE for the field trial, which was performed by Martin Porchas, coordinated by Sonnet Nelson and overseen by Paul Brierley. Martin's professional development included additional experience and knowledge gained from experiment station personnel, making him more proficient at melon field trials. California- A Laboratory Assistant I (Herman Daniel Delgado) has increased his familiarity with melon quality evaluation. Georgia- A student, Charle Proudfoot, participated in the project as part of a young scholars program at The University of Georgia. Information generated from this project will also be used for grower meetings during the winter of 2020. North Carolina- There were six undergraduate or post baccalaureate students, and one Research Associate involved in the 2019 study. Indiana- The project trained 10 undergraduate students and one technician about melon production, harvest and quality measurement. Texas - Uvalde: Trained 1 undergraduate student in melon field production; Trained 1 undergraduate student in melon quality evaluation; one graduate student obtained data for their MS thesis. Texas -Weslaco: 2 undergraduate student interns and two ag technicians were hired/trained to assist with project activities. Objective 4.Seven research technicians, three graduate students, and nine undergraduate students were trained in numerous techniques and skills related to food safety research, including but not limited to, sampling of melons and environmental samples, measurement of bacterial attachment strength, assessment of transfer rate of pathogens, antimicrobial treatments, biophotonic imaging, microbiome analysis and sample preparation for scanning electron microscopy. Four research technicians were provided an opportunity to train several undergraduate students in these and other techniques, thus providing them with development of supervising and training skills. Additionally, the research technicians, undergraduate and undergraduate students involved in the project are getting trained in field sample collection including melons and environmental samples. The SEM expert/scientist explored new imaging and analysis tools for surface characterization. Objective 5.Six undergraduate students received experiential learning opportunities in the areas of melon nutrition, analytical skills and working stakeholders at the Texas A&M's National Center for Excellence in College Station. Also, three undergraduate students were exposed to experiential training in the areas of food safety and collecting field samples from melon producers. How have the results been disseminated to communities of interest?Objective 1.Patil has presented research outcomes as invited/plenary/keynote speaker in several national and international conferences organized by American Chemical Society (ACS), American Society for Horticultural Science (ASHS), American Council for Medicinally Active Plants (ACMAP), Cucurbits symposium as a part of the International Society for Horticultural Science (ISHS) and Indian Horticulture Congress. Objective 2.Two seed company PD's observed melons in the Weslaco field plots during the Spring 2019. Student Fix presented two posters about the melon breeding project, one at the Cucurbitaceae conference at Davis, CA and one at the ASHS conference at Las Vegas. Objective 3. Arizona- The YCEDA Annual Report contained an item on the melon project. 400 copies were published and distributed at industry conferences and meetings, as well as via website and social media; The YCEDA website has a melon project description; Progress in the field trials, such as planting and harvesting, was broadcast to producers and academics via our social media channels. Georgia - The results have recently been compiled but will not get disseminated to growers until winter meetings. Two years of trials complete results will be disseminated in the form of extension publications. Silva presented the results in the ASHS conference. One popular press article has been published (http://www.caes.uga.edu/news-events/news/story.html?storyid=7645&story=Cantaloupes). This story has been picked up by several online outlets that reach vegetable growers. North Carolina - The information reported for 2018 can be found at the following URL and is available 24/7. https://cucurbits.ces.ncsu.edu/wp-content/uploads/2019/03/2018-North-Carolina-Orange-Flesh-Melon-Cultigen-Evaluations-Horticulture-Series-227.pdf?fwd=no Indiana - Through presentation at growers' meetings, extension publication and field days. (1) Results of the 2018 melon variety trial was presented at several growers' meetings. Each of presentations attracted 20-50 growers and agricultural professionals. Southwest Indiana Melon and Vegetable Growers Annual Winter Technical Meeting 2018 watermelon and Cantaloupe variety evaluations in Southwest Indiana. Vincennes, IN, Nov. 2018. Southern Illinois Fruit and Vegetable School. Cantaloupe varieties. Mt. Vernon, IL, Feb. 2019. Annual Watermelon Growers Meeting in Missouri. Watermelon and cantaloupe varieties. Kennett, MO. Dec. 2018. 2019; Indiana Hort Congress Watermelon and cantaloupe varieties. Indianapolis, IN, Feb. 2019; 2. Southwest Purdue Agricultural Field Day was conducted on June 27, 2019, which attracted about 85 growers and agricultural professionals. Melon trial was included in the field day. Texas -Uvalde and Weslaco: - Results of the project were disseminated via field day and professional meeting presentations: A spring vegetable field Day was held in Uvalde, TX on May 10th 2019 attracting more than 70 agricultural producers, gardeners, researchers, students and others. Results were also presented at: (1) the Annual Conference of the Texas Organic Farmers & Gardeners Association, Feb. 14 - 16, 2019 in Corpus Christi, TX, (2) the 83rd Annual Conference of the Southern Region of the American Society for Horticultural Science, Jan. 31-Feb. 5, 2019 in Birmingham, AL, and (3) the 116th Annual Conference of the American Society for Horticultural Science, July 21-25, 2019 in Las Vegas, NV. Objective 4.The results from the prevalence studies on Arizona melons, and the other food safety work including attachment strength of pathogens, cross-contamination potential of soil and dust, shelf-life studies, and imaging of melons have been presented in the form of posters at the Annual Meeting of the International Association for Food Protection at Louisville, KY in July 2019. The project was also demonstrated to produce industry professionals from Yuma, scientists and investigators from Arizona, food science professionals in Arizona and high school students from various areas in Arizona who toured the Ravishankar laboratories in 2018 and 2019. Objective 5.Results of the first year were presented to the Advisory Board (Principal Stakeholders and Scientific Managing Team) by each team lead leaders. Priority areas and potential impacts were discussed at length. What do you plan to do during the next reporting period to accomplish the goals?Objective 1. Phytochemicals such as carotenoids, vitamin C, phenolics, amino acids and aroma volatile compounds from two commercial varieties (Tuscan and F-39), five hybrids and various breeding lines grown in Weslaco and Uvalde will be analyzed. Furthermore, antioxidant activity and polyphenol oxidase using different in-vitro assays from these varieties will be determined. In addition, postharvest storage study of selected varieties of melons to understand the shelf life will be conducted. Objective 2. Additional seed will be produced from the top ten F1 hybrids by controlled pollinations in the greenhouse at College Station during the Fall 2019. This seed will be distributed to the production groups for planting in the Spring 2020 and also to several industry partners (growers) in Texas, Arizona, North Carolina and California. Controlled inoculations with Monosporascus will also be conducted on selected families to determine genetics of resistance and conduct SNP development. A second year of data will be collected from these field trials to write several refereed manuscripts, including a cultivar release document. Data will also be presented at the ASHS conference in Orlando. We will use customized RNA-seq library preparation kit (Universal Plus mRNA-Seq), which includes TRSV depletion module. This will eliminate majority of viral fragment in the RNA-seq library and increase the melon transcriptome information. The new melon samples are being harvested with fruit IDs that can match the individual RNA-seq data with phytochemical data without combining/averaging multiple fruits. This will increase the number of datapoints usable for the downstream association analyses. Objective 3. Arizona -Selected novel hybrids and breeding lines from the National Center of Excellence will be grown in the next season and evaluate both yield and quality and food safety characteristics. California - The fruit will be evaluated at maturity for yield and quality in 2019. At fruit maturity, there will be an extension meeting for growers and industry representatives to look at the trial. After harvest, field assessment data will be compiled and along with the 2018 data, the information will be extended to clientele at a meeting in December. Georgia-The plan prior to the next reporting period is to enter and statistically analyze the yield and quality data collected from the 2019 melon field study. We plan to produce a similar in-house publication that contains yield and quality information as was done in 2018. North Carolina - The plan prior to the next reporting period is to enter and statistically analyze the yield and quality data collected from the 2019 melon field study. We plan to produce a similar in-house publication that contains yield and quality information as was done in 2018. Indiana- Inform growers of the results in the second year at several growers' meetings as well as publish the result on Midwest Vegetable Trial Report and through our Newsletter, Vegetable Crops Hotline. Texas (Uvalde)- Selected novel hybrids and breeding lines from the National Center of Excellence will be grown in the next season and evaluate both yield and quality and food safety characteristics. Texas (Weslaco): Selected novel hybrids and breeding lines from the National Center of Excellence will be grown in the next season and evaluate both yield and quality and food safety characteristics. Objective 4. We will continue all the food safety studies on melons grown in various states including attachment strength of pathogens, measurement of melon rind surface roughness using SEM, cross-contamination potential of environmental matrices, natural interventions for melon decontamination, shelf-life studies, microbiome studies and bio-photonic imaging. We will also analyze melons grown in AZ and CA as well as the respective environmental matrices for presence of pathogens and indicator organisms. Objective 5. Plan to have Stakeholder Engagement Conference by coinciding with field day during June-July 2020. Develop bilingual short course material for Master Gardeners. Select specific undergraduate students who are interested in experience learning opportunities both in the lab and gain industry experience.

Impacts
What was accomplished under these goals? Objective 1. Texas A&M University provided seeds of the six hybrids and six cultigens; Melons were shipped from 7 different locations (GA, TX-Uvalde and Weslaco, NC,Indiana, AZ and CA) to TX to conduct flavor, sensory and quality including health promoting studies as well as to AZ to conduct fruit safety analysis. Brix from F-39was higher than other melons. In addition, Tuscan Davinci (TT-DV) from TX-Uvalde and Weslaco and NC showed higher brix. Four carotenoids were identified and quantified. Variation ofvitaminc C and amino acids were noticed amonng varities. Multiple factor analysis revealed the relationship between sensory, consumer attributes, sucrose, aroma compounds and phytochemicals. Key aroma related components and carotenoids derived volatiles were selected to understand the consumer and sensory panel liking. The highest consumer liking fruits are Harper type, TT-DV, F-39, Summer Kiss and Sugar Kiss. These fruits were positively correlated to ethyl acetate, but good floral and fruity aroma and β-ionone, were negatively correlated.Lowest consumer liking attributes such as musty/earthy, green and hardness were closely related to green woody aroma. Esters which impart fruity aroma and flavor were correlated with overripe, fermented, musty, earthy. Green and hardness attributes, due to certain aldehydes, were closely related to HD-150. Sour and bitterness were due to volatile compound like acetic acid, which were in close proximity for F-39 from AZ and IN, Harper type (HT-IG) from AZ, IN, TX-Weslaco; Carribean King from CA, OC-164 from CA and NC. The overall liking of the melon cultivars ensured from the combination of certain preferred primary aromatic, esters and carotenoids derived compounds. Breeding Lines. A total of 63 melon breeding lines were analyzed for volatile and phytochemicals compositions. Esters and few aldehydes and alcohols seems to be responsible for the flavor, aroma, anti-bacterial, anti-microbial activities of musk-melons. 39 were found to be more than 10 brix and few selected lines contains higher levels of volatiles related to aroma. Anti-bacterial and anti-fungal volatiles were found in BL 140, BL 99 and F-39. The breeding lines, BL-18 have higher yield, and low cucumber related odor compounds. Carotenoids content varied among the breeding lines. Variation of Citrulline GABA and tryptophan was demonstrated among breeding lines. Objective 2. A total of 46 novel melon F1 hybrids were produced by controlled pollinations. Twelve hybrids and 40 elite parents lines were planted for evaluation of important traits. Resistance to powdery mildew Monosporascus root root/vine decline and Alternaria leaf spot were observed but Cucurbit yellow stunting disorder virus was a big problem. Maturity ranged from first early (equal to Primo), to very late. Sugars measured as TSS ranged from 8 to 15 and flesh firmness from 18 N to 98 N. Fruit sizes ranged from 15 to 6, with most lines producing desired 9-12 sizes. Flesh thickness was equal or superior to Primo for more than 90% of the TAMU parents and hybrids. Flesh samples for volatile analyses of superior hybrids and inbreds were provided to the VFIC in the Spring. At Uvalde, 24 new F1 hybrids and their parents were grown to evaluate fruit quality and heritability for important fruit traits. The industry standard of TSS over 10 was found in 15 hybrids and six parents on average, with some individual fruits having over 14 TSS. Extreme flesh firmness over 60 N was found in 9 hybrids and 6 parents. Narrow sense heritability values were moderately high (>0.6) for flesh color, flesh firmness, fruit weight, fruit size and netting. Gene Expression Analyses. Twenty-one melon fruit samples have been processed for RNA-seq analysis. RNA extraction from mature fruits was optimized and uses Trizol extraction and purification using silica membrane column. This method successfully purified total RNA suitable for RNA-seq analysis from fruits with high water and sugar content. The sequencing quality of the samples checked using FastQC was reasonably good and different sample types are similar. The RNA-seq data were then aligned to melon reference genome CM4. 0.gff3. This is produced with double-haploid line DHL92, representing 83.3% of the estimated melon genome. They predicted 27,427 protein-coding genes. Of trials using different packages, STAR produced the best results. Initial alignment resulted in large proportion of unmapped reads, which turned out to originate from TRSV (tobacco ringspot virus). The viral sequences were filtered to produce each sample dataset for further analysis. Expressions of approximately 22,000 unique melon transcripts were detected by the RNA-seq analysis. Separately, viral sequence was assembled to produce full-length viral genome sequence to identify strain of detected virus. Approximately 200 genes whose expression is highly correlated (P<0.01) with the sensory score were selected for further analysis. Objective 3 Selected melons from each cultivar/hybrids/breeding lines were shipped from each state to Texasfor quality and sensory/genomics analyses and to Arizonafor food safety investigations. Fruit counts, weights, dimensions, solids and firmness were recorded. Yields, in marketable fruit, cartons and total tonnage production were also calculated. Fertilization, management and IPM are consistent with commercial practice. Additionally, Weslaco field trials with 140 advanced melon breeding lines, a pre-commercial trial of newly developed western shipper hybrid melons as well as 24 F1 hybrids testing in Uvalde were conducted. Objective 4.The enterococci population and Escherichia coli population for melons soil, rhizosphere, air and water samples, were measured. The attachment strength of Salmonella ranged from 0.07-0.53 and that of Listeria monocytogenes ranged between 0.13 and 0.43. Some melons that had a high netting % area based on surface netting analysis also showed higher values for attachment strength for Salmonella. The melon varieties also were sampled for enumeration of total plate count, coliforms count and yeast and mold count on days 0 and 3. The varieties with the highest microbial load were F39 and Davinci, while the lowest microbial load was found on varieties Honeydew 252 and OC 164. Using the heat map, it was seen that not a significant loss of shelf-life was observed. Infinite gold of AZ had the highest and HD252 from NC had the lowest plate count on day 3 of storage. DaVinci Sakata from CA and GA had the most coliforms and fungal growth, respectively, on day 3. Cantaloupes in general harbored more microbes than honeydews. Cross-contamination study revealed that soil showed a higher % transfer than dust, and cantaloupes in general had a higher % transfer than honeydew melons. Percent transfer increased with inoculum level in soil. The effectiveness of plant antimicrobials (5% olive extract and 0.5% oregano oil) in wash water for inactivating Salmonella and L. monocytogenes was investigated. Compared to control wash (PBS), the antimicrobial treatments caused an additional 2-3.6 and 1.6-3.7 log reductions on Salmonella and L. monocytogenes, respectively. In general, oregano oil had better antimicrobial activity than olive extract. The antimicrobial treatments were more effective on Salmonella than on L. monocytogenes. The antimicrobial treatments were more effective on honeydews than on cantaloupes. Biophotonic imaging of inoculated cantaloupe rinds with various netting patterns showed that dense netting harbored more bacteria in comparison to medium and low netting. Also, bacteria tend to attach more on the netting than on the grooves. Hence, the denser the netting, the more surface area will be available for bacterial attachment.

Publications

• Type: Journal Articles Status: Published Year Published: 2019 Citation: Corleto, K. A., Singh, J., Jayaprakasha, G., & Patil, B. S. (2019). A sensitive HPLC-FLD method combined with multivariate analysis for the determination of amino acids in L-citrulline rich vegetables. Journal of Food and Drug Analysis, 27(3), 717-728.
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Kasote, D. M., Jayaprakasha, G. K., & Patil, B. S. (2019). Leaf disc assays for rapid measurement of antioxidant activity. Scientific Reports, 9(1), 1884.
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Kasote, D. M., Lee, J. H., Jayaprakasha, G.K., & Patil, B. S. (2019). Seed Priming with Iron Oxide Nanoparticles Modulate Antioxidant Potential and Defense-Linked Hormones in Watermelon Seedlings. ACS Sustainable Chemistry & Engineering, 7(5), 5142-5151.
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Perez, J. L., Jayaprakasha, G.K., & Patil, B. S. (2019). Metabolite profiling and in vitro biological activities of two commercial bitter melon (Momordica charantia Linn.) cultivars. Food Chemistry, 288, 178-186.
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Perez, J. L., Jayaprakasha, G. K., Crosby, K., & Patil, B. S. (2019). Evaluation of bitter melon (Momordica charantia) cultivars grown in Texas and levels of various phytonutrients. Journal of the Science of Food and Agriculture, 99(1), 379-390.
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Shivanagoudra, S. R., Perera, W. H., Perez, J. L., Athrey, G., Sun, Y., Jayaprakasha, G., & Patil, B. S. (2019). Cucurbitane-type compounds from Momordica charantia: Isolation, in vitro antidiabetic, anti-inflammatory activities and in silico modeling approaches. Bioorganic Chemistry, 87, 31-42.
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Shivanagoudra, S. R., Perera, W. H., Perez, J. L., Athrey, G., Sun, Y., Wu, C. S., Jayaprakasha, G., & Patil, B. S. (2019). In vitro and in silico elucidation of antidiabetic and anti-inflammatory activities of bioactive compounds from Momordica charantia L. Bioorganic & Medicinal Chemistry, 27(14), 3097-3109.
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Singh, J., Metrani, R., Shivanagoudra, S. R., Jayaprakasha, G. K., & Patil, B. S. (2019). Review on Bile Acids: Effects of the Gut Microbiome, Interactions with Dietary Fiber, and Alterations in the Bioaccessibility of Bioactive Compounds. Journal of Agricultural and Food Chemistry, https://doi.org/10.1021/acs.jafc.8b07306.
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Wang, J., Jayaprakasha, G.K. & Patil, B. S. (2019). Chemometric characterization of 30 commercial thermal and cold processed juices using UPLC-QTOF-HR-MS fingerprints. LWT, 114, 108322.
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Acharya P., Jayaprakasha, G.K., Crosby, K., Jifon, J., and Patil, B.S. (2019). Green-Synthesized Nanoparticles Enhanced Seedling Growth, Yield, and Quality of Onion (Allium cepa L.). ACS Sustainable Chem Eng. https://doi.org/10.1021/acssuschemeng.9b02180
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Fix, A., Patil, B., Leskovar, D., Rooney, W., Smith, W., and K. Crosby (2019). Mission melon: improving qualitative traits in Cucumis melo using phenomics. Annual Conference of ASHS, Las Vegas.
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Wei, Q., M. Torres, M. Porchas, T. Fang, B. Patil, P. Brierley, and S. Ravishankar (July 2019). Attachment Strength of Foodborne Pathogens on Different Melon Varieties from Various Regions in the United States. Poster presented at the Annual Meeting of the International Association for Food Protection. Louisville, KY.
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Park, R., A. Rao, M. Porchas, B. Patil, P. Brierley, and S. Ravishankar (July 2019). Understanding Cross-Contamination of Melons via Environmental Matrices Under Field Conditions. Poster presented at the Annual Meeting of the International Association for Food Protection. Louisville, KY.
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Chun-Chi Chen, Hyundoo Jeong, Xiaoning Qian, Byung-Jun Yoon, "TOPAS: network-based structural alignment of RNA sequences, Bioinformatics, in press, doi: 10.1093/bioinformatics/btz001, 2019.
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Rao, A., M. Porchas, B. Patil, P. Brierley, and S. Ravishankar (July 2019). Determination of the Shelf-Life and Keeping Quality of Melons Grown In Different Regions With Respect To Background Microflora, Prevalence of Foodborne Pathogens and Variances in Netting Densities. Poster presented at the Annual Meeting of the International Association for Food Protection. Louisville, KY.
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Shivanagoudra, R.; Perera, W. H.,; Perez, J. L.; Giridhar, A.; Sun, Y.; Wu, C.S.; Jayaprakasha, G. K.; Patil, B. S. (April 8-11, 2019). Cucurbitane type-triterpenes from bitter melon inhibit ?-amylase and ?-glucosidase and display in vitro anti-inflammatory Activities, 19 Annual Oxford International Conference on the Science of Botanicals.
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Patil, B.S., Singh J., Acharya, P., Metrani, R., Shivanagoudra, S. R, Perez, J. L., Jayaprakasha, G.K. (June 30 -July 4, 2019). Hidden treasures of the Cucurbitaceae: Health benefits of melons, from watermelon to bitter melon. ISHS, VI International Symposium on Cucurbits, Ghent, Belgium.
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Patil, B.S., Jayaprakasha, G.K., Singh J. (Feb 14-16, 2019). Fruits, vegetables, and medicinal plants for the prevention and treatment of disease: untapped potential and unanswered questions, 9TH ACMAP Conference  Dehradun, India.
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Patil, B.S. Horticulture, nutrition, and human health: status, challenges and prospects for implementing a sustainable diet (Jan 17-21, 2019). 8th Indian Horticulture Congress (IHC), Raipur, Chhattisgarh, India.
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Patil, B.S., Jayaprakasha, G.K., Singh J. (July 21 to 25, 2019). Optimization of factors influencing quantitation of carotenoids of melon. ASHS Annual Conference - Las Vegas, NV.
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Metrani, R., Patil, B.S., Jayaprakasha, G.K. (March 31- April 4, 2019) Potential of aroma volatile compounds to improve the melon shelf-life and food safety. 257th American Chemical Society National meeting & Exposition, Orlando, Florida, USA.
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Jifon,J., Patil, B.S., K.C. Crosby (July 21 to 25, 2019). Muskmelon (Cucumis melo L.) Yield and Quality Responses to Potassium Fertilizer Sources. 116 ASHS Annual Conference - Las Vegas, NV.
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Perera, W. H., Shivanagoudra, R.; Perez, J. L.; Sun, Y.; Wu, C.S.; Jayaprakasha, G. K.; Patil, B.S. (July 13-17, 2019). Anti-inflammatory and carbolytic enzymes Inhibitory activities of monoglycoside Cucurbitane-type triterpenes from Momordica charantia, 2019 Annual Meeting of the ASP, Madison, MO.
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Jifon, J., Crosby, K., Patil, B., Leskovar, D. (2019). Yield and quality of muskmelon (Cucumis melo L.) as affected by mineral nutrition. Presentation at the 83rd Annual conference of the Southern Region of the American Society for Horticultural Science, Jan. 31-Feb. 5, 2019, Birmingham, AL.
• Type: Other Status: Other Year Published: 2019 Citation: Rao. A. P. and Ravishankar. S. Presentation titled Cleaner and Greener Alternatives for Safe Food- Its Essential! to Environmental Health Inspectors at the Maricopa Environmental Health Services Department, Phoenix, AZ, April, 2019.
• Type: Other Status: Other Year Published: 2019 Citation: Rao. A. P. and Ravishankar. S. Presentation titled Cleaner and Greener Alternatives for Safe Food- Its Essential! to the local high school students and their mentors at the USDA-Ag Discovery Camp, UA- Tucson, AZ, June, 2019.
• Type: Other Status: Other Year Published: 2019 Citation: Ravishankar, S. Presentation titled Cleaner and Greener Alternatives for Safe Food- Its Essential! to university students, scientists and professors at the Sathyabama University, Chennai, India, June 27, 2019.
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Andre Luiz B.R. da Silva, Timothy W. Coolong, Kevin Crosby, and Bhimanagouda S. Patil (July 21 to 25, 2019). Presentation titled Cantaloupe and Honeydew Yield and Quality Performance in Georgia ASHS Annual Conference 2019
• Type: Other Status: Published Year Published: 2019 Citation: Egel, Dan and W. Guan. 2019. Evaluation of melon varieties for resistance to bacterial wilt, 2018. Plant Disease Management Reports. 13:V057.

Progress 09/01/17 to 08/31/18

Outputs
Target Audience:The transdisciplinary team from seven states of Mission melon project targeted stakeholders including scientific colleagues, melon growers, retailers, shippers, distributers, food safety managers, and other produce industry professionals. Stakeholders were informed about the project objectives and were asked to participate by giving their input on project directions, so that the focus is on industry needs. They were also requested to provide commercial melon fruits for prevalence studies. Project investigators from different disciplines involving 'consumer to farm' aspects provided training to postdoctoral researchers, students and field technicians in breeding, crop management, sensory, flavor, nutrition, genomics, bioinformatics, analytical skills, food safety and consumer acceptance. Project investigators in seven states reached and/or planning to reach to melon industry members in their respective states by presenting their results in extension meetings. Demonstrated the food safety aspects of the melons project currently conducted in University of Arizona to a group of 30 high school students and their mentors from various regions throughout the state of Arizona, participating in the USDA-Ag Discovery Camp, on June 25, 2018. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Objective 1. A postdoctoral researcher, a research associate, a graduate student, and 4 undergraduate students have been responsible for quality, flavor and health promoting compounds. The students received training in numerous advanced techniques and skills related to melon sensory, extraction, instrumentation and analytical skills. We also developed "train-the-trainer" concept to continue professional development for future generations. Objective 2. Two graduate and two undergraduate students have been trained in melon breeding protocols, and one graduate student trained in RNA extraction and RNA-seq. Another Ph.D.has been recruited to work to develop Bayesian differential expression and factor analysis methods for RNA-seq data to identify candidate genetic markers and factors for the phenotypic outcomes of interest. Objective 3. In each of partner states, several talented employees including graduate and undergraduate students were trained on the required skills in relation to crop management, and provided opportunities for professional development. Arizona - Farm Attendant is hired to oversee the field trial in Yuma. He also assisted in the planning of a Melon Research Field Day. Georgia - A student participated in the project as part of a young scholar's program at The University of Georgia. Information generated from this project will also be used for grower meetings during the winter of 2019. An agent training (extension agents) was conducted on 15 June and this project was included in the training. North Carolina - Four summer interns and one graduate student worked on this project and assisted with all aspects of trial establishment, maintenance and harvesting. Indiana- 5 undergraduate students participated in the trial. They received training in melon production. Results of the experiment will be shared with Indiana melon growers as well as seed company representatives at the Southwest Indiana Melon & Vegetable Growers Winter Meeting. Texas - Uvalde: The trial was highlighted during the spring vegetable field day which allowed participants to learn about difference in agronomic traits of melon varieties. Students working during the summer break were also involved in the trial management allowing them to also learn about techniques of melon production. Texas -Weslaco: Two undergraduate student interns and two ag technicians were hired/trained to assist with project activities. Objective 4. One research specialist, 3 research technicians, 2 graduate students, 9 undergraduate students and 3 high school students worked in the Food safety lab in University of Arizona. They learn numerous additional techniques and skills related to extraction, instrumentation, food safety research, such as biofilm formation, antimicrobial treatments, microbiome analysis and scanning electron microscopy sample preparation. Furthermore, it has also provided the opportunity for the research technicians and graduate students to train undergraduate and high school students in these and other techniques, thus providing them with the experience of supervising and training students in the laboratory. Students are also getting trained in team work, protocol development, and data analysis Objective 5. One PhD student joined the project and continued initial work and the survey. The survey was administered online using Qualtrics and was made up of a combination of rating scales and open-ended items that sought to gather participant's perceptions in retail quality, safety and nutrition quality of melons. The student has been trained in data analyses using STATA and SPSS statistical packages. She also learned how to prepare power points and posters for presentation at peer reviewed conferences. How have the results been disseminated to communities of interest?The project investigators disseminate results to their respective state communities, industry stakeholders and provide information to develop national level thematic messages through National Center of Excellence. On June 19, 2018, the project PD and team members from TAMU and UA gave presentations at Yuma, Arizona on the project objectives to the melon industry professionals (growers, distributers, food safety managers, etc.) in Arizona and California. Field Demonstration Day to showcase the new varieties of melons grown in the plots at the Yuma Ag Center, Yuma, AZ was also organized by the co-PI in Arizona. Objective 2. Two posters and one field day presentations were produced by co-PI and graduate student. Two poster presentations related to pest control in melon were presented to key melon industry and administrators of Texas A&M AgriLife. California - The melons in the field trial is at early stages of development and information has not been disseminated to growers. There are plans to extend the information generated in a meeting held at the University of California West Side and Extension Center in Fresno County California. Georgia - The results have recently been compiled but will not get disseminated to growers until winter meetings. Then after two years of trials complete results will be disseminated in the form of extension publications. An agent training was conducted on 15 June and this project was included in the training. Approximately 24 extension professionals attended the training. The young scholar already presented preliminary results at a student research forum this summer, but our primary community of interest is growers, which will be reached through other mechanisms. One popular press article has been published (http://www.caes.uga.edu/news-events/news/story.html?storyid=7645&story=Cantaloupes) This story has been picked up by several online outlets that reach vegetable growers. North Carolina - Statistical analysis of the trial data is currently underway. Trial results will be presented in booklet form as an extension publication. In addition to the seven cultivars/varieties of this trial we evaluated 17 other melon entries for evaluation. The results will be presented as a slide show presentation at various extension meetings/trainings and grower associations with specific melon production interests. Indiana - The results will be complied and shared with growers in the winter meetings. Texas - Uvalde: A spring vegetable field day was held on May 11, 2018 at the center. Goals and specific objectives of the project were discussed with the participants during the field tour. Texas - Weslaco: A mid-season field day was held on April 13, 2018 including growers and seed/agrochemical company representatives to discuss project goals and anticipated impacts for consumers and the retail industry. Objective 4. Food Safety team leaders toured labs and demonstrated the food safety aspects of the melons project to a group of 30 high school students and their mentors from various regions throughout the state of Arizona-USDA-Ag Discovery Camp, June 25, 2018, UA-Tucson, AZ. Objective 5. A total of 235 stakeholders completed an anonymous Qualtrics survey. A manuscript entitled Perceptions of Quality, Safety, and Attributes of Cantaloupe and Honeydew Melons by Consumers and Industry Representatives was submitted to the International Journal of Vegetable Science and is currently under second review. A proposal from the results of the study has been submitted to the Academy of Human Resource Development Annual Research Conference to be held in February 2019. Results have also been shared by the key stakeholders. What do you plan to do during the next reporting period to accomplish the goals?Objective 1. During the next year, PD's lab will analyze the variation in the health promoting compounds and flavor related compounds such as carotenoids, volatiles, ascorbic acid, amino acids, and phenolic acids in the different melon breeding lines and cultivars. Effect of post-harvest storage on the health promoting compounds will also analyzed. Also, antioxidant activity using different in-vitro assays will be determined. The data will be completed for the trained and consumer sensory work. The data will be analyzed and varieties within states will be selected for use in Objectives 2, 3 and 4. Objective 2. Distribute seed of 25-30 new F1 melon hybrids to seven locations across the U.S. Compile phenotypic data for all quality and performance traits. Determine genetic components and metabolomic markers for the fruit quality traits impacting flavor and food safety. The published manuscripts listed are for developing general methods to analyze RNA-seq data. During the next year, the developed methods can be used to analyze melon RNA-seq data. Objective 3. Arizona - We will obtain melons from commercial fields along with correlating soil, water and air samples and send to University of Arizona for laboratory sampling and microbial analysis. California - By the next reporting period, this trial will be completed, the data will be tabulated and analyzed and will have been presented at extension meetings and in an online publication through University of California Cooperative Extension. The second season trial will be in progress. Florida - Repeat the trial as per grant objectives, tasks and disseminate information to grower groups during winter meetings. Planting date should be postponed 1-2 weeks. Georgia - Repeat the trial as per grant objectives and disseminate information to grower groups during winter meetings. North Carolina - Information obtained from the trial will be summarized in our annual melon cultivar evaluation booklet (extension publication), as well as, being presented at future extension meetings or to other melon associations in winter, 2019. Indiana- The project results will be shared with growers and seed company representatives at the upcoming vegetable growers' winter meetings. The information will also be incorporated in the 2019 edition of the Midwest Vegetable Production Guide. A newsletter article about melon production will be published at the Purdue Extension Newsletter Vegetable Crops Hotline. Texas - Uvalde and Weslaco: In both locations, second season investigations on GxExM effects on melons productivity and quality as well as second season screening of advanced germplasm from Objective 2 will be conducted. Objective 4. Food safety related studies will continue as per for the time line and tasks listed in the proposal including measurement of attachment strength of foodborne pathogens/biofilm formation on melon rinds and surface roughness of melon rinds from various cultivars; role of soil and dust in cross-contamination of field crop; efficacy of plant antimicrobial interventions on foodborne pathogens on melon rinds; shelf-life of melon cultivars under various storage conditions; and biophonic imaging of artificially contaminated melon rinds to understand areas prone to attachment and contamination. Evaluate the surface roughness of melon rinds correlates with bacterial attachment and biofilm formation. UA Food Safety lab will also continue the study on microbiome samples and conduct the data analysis for the commercially purchased melons and different melon cultivars. Additional work with different melon growers to set up in field sampling of melons for pathogen prevalence studies and in field melon microbiome analysis in Arizona and California. Objective 5. Tentatively, Stakeholders Engagement Conference is scheduled during 2019. The results of the survey of consumers and industry professionals will be shared. It is expected that all investigators will present their results to stakeholders from seven states. In addition, a survey regarding the production and consumption of safer and healthier melons will be administered prior to the conference and three months after the conference. Meeting is arranged with retailers to discuss about "retailer-melons for health" dialogue on nutrition and quality aspects to the selected grocery chain produce managers.

Impacts
What was accomplished under these goals? Objective 1: Considering the funding and setting up the account in each collaborating institutions took some time, this reporting period mostly provide details of initiation of experiments rather than results. Seven varieties of melons were grown in Texas, Georgia, North Carolina, California, Indiana, Florida and Arizona. Until now, melons were harvested and shipped to Texas A&M University from five states (TX, GA, NC, IN, and AZ) for evaluating the consumer acceptance, sensory qualities, and health-promoting phytonutrients. Texas A&M team (including sensory specialist, analytical chemist, natural product chemist, agriculture economist) are conducting analysis and studies related consumer acceptance, trained sensory evaluation, flavor and analysis of health-promoting compounds including aroma compounds responsible for food safety, quality parameters, ascorbic acid, amino acids, and phenolic acids. During the same time, equal amount of fruits were shipped to University of Arizona for testing of food safety attributes managed by the microbiologist. Due to weather problems, Florida melons could not be used for any analysis. Consumer acceptance test and economic potential The first part of the consumer study is currently undergoing. Subjects were recruited from the general population in College Station and Bryan, Texas. The experimental sessions with consumer groups were conducted followed by consumer experimental data from over 500 participants were collected. Quality, phytochemicals, flavor profiling, and sensory evaluation The melons from five states were evaluated for trained descriptive color, flavor and texture attributes using an expert descriptive panel with almost 30 years of experience. A melon lexicon was developed prior to evaluation of the melons that includes attributes, definitions and references. This lexicon will be used for the duration of the study. After trained descriptive evaluation, consumer sensory evaluation was conducted on the same varieties from the 7 states. Rapid and accurate methods for the measurement of carotenoids and vitamins by HPLC. Before analyzing the melon samples from 7 states, several parameters were optimized for carotenoids analysis such as blending time and extraction efficiency using different solvents. Results demonstrated that the best solvent for the extraction of carotenoids was chloroform: acetone (1:1) and ratio of MPA: IPA (1:1) for Vitamin C extraction. Variation of carotenoids and vitamin C in different locations. Initial results suggest variation of the carotenoids profiles of melons harvested from different locations from five states. The fruits of Tuscan Da-Vinci grown from Weslaco, Texas and Tifon, Georgia shows significantly higher levels of β-carotene and ζ-carotene compared to other locations. Similarly, Harper Type (infinity) harvested from Georgia showed significantly higher β-carotene than other locations. Objective 2. The melon breeding team has screened thousands of melon lines, cultivars and germplasm accessions for durable resistance to multiple diseases, stresses, and product quality. Three field trials with elite breeding lines and hybrids have been conducted in Texas at Weslaco, Uvalde and College Station during the first year. Seed for more than 30 new F1 hybrids and about 100 parent lines have been produced in greenhouse and field nurseries. Two graduate students and two undergraduate students have been trained in melon breeding protocols. A total RNA extraction method was established for melon fruits. We found combination of Trizol extraction and subsequent column purification produced sufficient yield and quality of total RNA from melon fruit of various maturities. Using the optimized extraction method, triplicate RNA samples were prepared from 7 varieties grown in Texas. Qian's group has recently developed a new suite of Bayesian differential expression analysis methods (see the publication list 1-2) that achieve significantly better differential expression results on benchmark RNA-seq data sets by avoiding the heuristic data normalization steps in existing differential expression methods, including Limma, EdgeR, and DESeq2. Objective 3. Each participating states conducted field trail of seven commercial varieties/cultivar developed by Texas A&M breeding program including their respective commercial variety as control. Selected number of melons from each variety were also shipped to Texas A&M for nutrition, quality, and flavor analysis and to University of Arizona for food safety evaluation. Arizona - The Yuma Center of Excellence for Desert Agriculture conducted a variety trial with seven melon varieties. California - Seven melon entries are being compared in a June-planted trial in Fresno County California. Fruit quality will be assessed on site at maturity. Florida - The trial was in Plant City, Florida but due to unexpected weather, fruits from field trials could not be used for further analysis. Georgia - This trial was conducted in Tifton, GA in spring 2018. North Carolina - A variety trial was conducted using 7 varieties of melon with different agronomic characteristics in Clayton, NC. Fruit yield and quality (interior/exterior) were collected. Melons in this trial were evaluated for adaptability to North Carolina growing region, as well as, being compared to other cultivars already commercially produced in the region. Indiana - A trial comparing the 7 melon varieties was conducted at the Southwest Purdue Agricultural Center in Vincennes, IN in Spring Season 2018. The trial evaluated yield, fruit quality and disease resistance of the 7melon varieties. Texas - Uvalde: Yield performance and basic quality attributes of seven varieties of melon were evaluated in an open-field trial at the Texas A&M AgriLife Research Center in Uvalde, TX. Texas - Weslaco: 2 field trials were completed (1) a germplasm screening trial including over 150 advanced melon hybrids and (2) a variety trial with 7 commercial and pre-commercial entries was conducted to characterize productivity and consumer/retail quality traits under south Texas growing conditions. Objective 4. Seven varieties of melons from 6 locations were received. These melons are being evaluated for safety and shelf-life. The attachment strength of foodborne pathogens on these melon rinds is measured and samples from the same melons are preserved in methanol for measuring the surface roughness of the rinds. The role of soil and dust in transferring foodborne pathogen contamination to rind surface and the efficacy of plant antimicrobial interventions in decontaminating the rind surface is also evaluated. The shelf-life of whole melons at refrigeration and abuse temperatures is analyzed. We have begun to examine the microbiome of various types of commercial melons at the point of consumer purchase to understand those bacterial communities that are present on the different melon surfaces at time of purchase. Objective 5: The National Center of Excellence of Melon was officially launched and an advisory board consists of a diverse stakeholder advisory panel and managing scientific team is formed. Stakeholder panel include members from each melon growing states and representatives from producers, produce associations, seed companies and retailers. First meeting of advisory meeting was conducted and assessed project progress and impacts, and planned for future activities. Project website https://missionmelons.tamu.edu/ is launched, project accomplishments and products will be posted as we continue the progress of the project. Communicated with CuCAP project director and discussed about the possibility of sharing their relevant project results which may have mutual interest to our investigators and eventually benefit the melon stakeholders and consumers.

Publications

• Type: Journal Articles Status: Awaiting Publication Year Published: 2018 Citation: Siamak Zamani Dadaneh, Mingyuan Zhou, Xiaoning Qian. Bayesian negative binomial regression for differential expression with confounding factors, Bioinformatics, in press, DOI: 10.1093/bioinformatics/bty330, 2018.
• Type: Journal Articles Status: Awaiting Publication Year Published: 2018 Citation: Siamak Zamani Dadaneh, Mingyuan Zhou, Xiaoning Qian. Covariate-dependent negative binomial factor analysis of RNA sequencing data, Bioinformatics, 34(13): i61-i69, 2018. PMID: 29949981.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2018 Citation: Patil, B.S., S.V. Angadi, and Senthold, A. 2018. Exploring Modeling to determine Environmental Conditions effect on health promoting components of melons. 7th AgMIP Global workshop, San Jose, Costa Rica. April 23-26, 2018.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2019 Citation: Mkuu, R., M.A. Palma, F. Nafukho., and B.S. Patil 2018. Perceptions of Quality, Safety, and Attributes of Cantaloupe and Honeydew Melons by Consumers and Industry Representative. Academy of Human Resource Development Annual Research Conference. Feb 2019.