Progress 08/15/20 to 08/14/24
Outputs Target Audience:In this project, we reached leafy green hydroponic producers, students, and researchers in the areas of controlled environment agriculture, plant pathology, microbiology, and food safety. Members of our team visited and collaborated with ten hydroponic leafy green facilities in the state of Ohio. During these visits, we learned about their production practices and obtained samples for the characterization of microbial communities on nutrient solutions, plant compartments, and other surfaces. We also obtained material from growing substrates and water used in the production practices. Specific results from individual production facilities have been shared with growers who participated in our project, and aggregated results have been presented to other industry collaborations through the Phytobiomes Alliance and the Ohio Controlled Environment Agriculture Center (OHCEAC). Phytobiomes Alliance and OHCEAC are industry-academia collaborative efforts that focus on system-level research in agriculture. Dr. Benitez Ponce is currently the co-chair of Phytobiome Alliance's coordinating committee of the Controlled Environment Agriculture Working Group and as part of that contributes to conversations about research needs within CEA microbiomes and to conference event organization. OHCEAC is an Ohio State University (OSU) industry collaboration with regular meetings with OSU researchers and industry partners which have provided opportunities to present results, obtain feedback, and fund additional research. Through our food safety component led by Drs. Ivey and Ilic, our team reached growers and personnel beyond our sampling efforts through online and in-person training modules and conferences. For example, updates on good agriculture practices to promote food safety in hydroponic productions were published, with an accompanying online self-paced course developed (in two languages). Results from our project were shared with the research community through oral and written communication, including invited seminars and webinars at various academic institutions and scientific societies, and through peer-reviewed manuscripts and manuscript pre-prints. We provided research training in hydroponic production, and microbiome analysis (wet-lab and in silico) to undergraduatestudents (OSU -3 , College of Wooster -1, University of Puerto Rico -1), visiting scholars (Mexico -1), Master (4) and PhD (3) students from Plant Pathology and Human Nutrition, postdoctoral researchers (4) and technicians (2). In addition, we developed teaching material that was integrated into the Advanced Plant Pathology laboratory course (PP6002.01/6010, Department of Plant Pathology at OSU) and the Introduction to microbiome workshops hosted in collaboration with the Molecular, Cellular and Imaging Center at OSU's Wooster campus. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?As part of our project, and to train project participants (post-doctoral researcher, research technician and graduate student) we developed a workshop on experimental design considerations and data analysis of amplicon sequencing experiments for plant microbiome analysis. This workshop was imparted twice during the duration of the project, with a total of 24 participants overall. In addition, this supported the professional development of one PhD student and a post-doctoral researcher as instructors, in charge of both developing and delivering content. Throughout the course of this project, undergraduates, graduate students, post doctoral researchers, research technicians and faculty members had opportunities to attend and present in scientific conferences, greenhouse trade meetings and academic seminars and workshops. Among them, members of our team presented and participated in the Ohio Controlled Environment Agriculture Center annual meetings on three years. Other scientific conferences that provided networking and career development opportunities including, the MidWest Microbiome Symposia, OSU's Infectious Disease Institute annual meetings, OSU's Center of Microbiome Science annual meetings, the annual meetings Plant Health Meetings, the International Association of Food Protection, OSU's Code Club. The work developed as part of this project was part of onecompleted undergraduate student internship project, two completed MSc student dissertations, and two PhD student dissertations. The project also provided opportunities to six postdoctoral researchers and research technicians. How have the results been disseminated to communities of interest?The results have been disseminated to both the scientific community and hydroponic producers. As part of this, several seminars at academic institutions and scientific meetings, as well as poster presentations and short talks are listed under Products. In addition, our team continues to develop online and printed resources about hydroponic production and food safety management in leafy green hydroponics, which are available through the team's websites: https://u.osu.edu/benitezponce-1/links/ and https://producesafety.osu.edu/. We also discuss aspects of this research with growers and industry affiliates through our interactions with OHCEAC and Phytobiomes Alliance. 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 characterized the diversity and relative abundance of bacteria and fungi in 10 commercial leafy green hydroponic facilities in Ohio. Microbial communities were analyzed from various hydroponic environments, including surface swabs, growing media, nutrient solution, lettuce leaves, and roots. Samples originated from facilities that varied in size (23-13150m2), system setup (Nutrient Film Technique (NFT), Deep Water Culture (DWC), Ebb & flow (EF), or Vertical Drip (VD)), water source used for nutrient solution preparation, as well as pH and EC parameters of nutrient solution. We applied amplicon metabarcoding of the ribosomal markers, 16S rRNA, and ITS1 for bacteria and fungi, respectively. Community composition analysis indicated that the bacterial and fungal communities' structure differed between habitats (leaves, roots, and nutrient solution). Bacterial and fungal richness and evenness also showed differences between habitats, with roots and nutrient solution showing higher richness than leaves. However, bacterial evenness was significantly higher in leaves, whereas fungal evenness was higher in leaves and nutrient solution, compared to roots. Further comparison between sampled compartments across facilities showed higher bacterial richness, but lower evenness in DWC systems, whereas greater fungal richness and evenness were observed in EF and NFT systems. Factors such as system type, plant age, nutrient solution parameters (pH, EC, and temperature), and facility contributed to the observed community variation. We identified members of the bacterial genera Flavobacterium, Sphingobium, Pseudomonas, and Rhizobium, and the fungal genera Olpidium, Golovinomyces, and Plectosphaerella as the most abundant and prevalent in the sampled facilities. Lastly, communities from nutrient solution were less variable across facilities, compared to other sample types and, consistent groups of bacteria were recovered from the nutrient solution (through amplicon metabarcoding and culturing), including Bosea, Sphingomonas, and Sphingobium. A collection of over 160 bacteria from the nutrient solution of the same commercial facilities was evaluated for biocontrol and growth promotion activity, including whole genome comparative analysis of 72 isolates. 32 isolates classified within 12 genera, including Pseudomonas, Sphingobium, Chryseobacterium, Bacillus, Nivierspirillum among others, showed biocontrol activity against at least one of the two plant pathogens tested. 8 and 46% of the isolates showed invitro phosphate solubilization and siderophore production. Analysis of whole genomes showed that isolates that inhibit both bacteria and oomycetes possess biosynthetic gene clusters (BGCs) to produce diverse antimicrobial compounds including hydrogen cyanide (HCN), brabantamide A and thanamycin. Genomic analysis also provides insights into functions associated with bacteria native to hydroponic systems. Objective 2. Our team performed a series of greenhouse experiments in research deep water cultures (DWC) and nutrient film technique (NFT) systems. We evaluated how system setup and nutrient solution management influenced bacterial and fungal communities in various hydroponic system compartments, as well as effect on plant growth. We manipulated electrical conductivity (EC), pH, and the water source. Experiments for EC and pH manipulation were completed in both experimental DWC and NFT; whereas water source experiments were completed in a small-scale, 5-gallon DWC setup. As expected, greater lettuce vigor was observed under recommended conditions (1.8 EC and pH of 5.8); but, we recovered more culturable bacteria from roots of lettuce grown at an EC of 2.5. From the pH experiments, we determined that maintaining a consistently low pH (5.0) is challenging and is related to changes in the nutrient solution as the lettuce head begins to form. This resulted in inconclusive data on pH effects on microbial diversity and abundance in system compartments. Lastly, lettuce vigor, when grown in a nutrient solution prepared with rainwater (RW), was higher, than when grown in a nutrient solution prepared with reverse osmosis-filtered municipal water. In addition, a subset of roots from lettuce grown in RW nutrient solution harbored diverse bacterial communities, which differed in dominant bacterial taxa, compared to municipal water. Plant and growth media samples from these experiments (nutrient solution management) have been processed for nucleic acid extraction and microbial communities were characterized through amplicon metabarcoding. We also tested the effect of spent, or reused, nutrient solution, and its microbial community, on subsequent lettuce growth cycles. We determined that a) nutrient solution can be used to seed, or inoculate, a leafy green hydroponic setup and b) the host plant drives the composition of newly setup system. Also, no significant effect on lettuce biomass was observed, regardless of nutrient solution origin and the richness of bacteria recovered from the nutrient solution was lower at the end of the experiment, compared to the original "seed". When considering other plant species in this experiment, we observed that plant species was a greater microbial community driver than water source (or spent nutrient solution origin). Lastly, we tested the influence of surface materials (i.e. polyethylene liner and PVC, for DWC and NFT, respectively) on microbial buildup in the form of biofilms. Our results indicated that the pond liner used in DWC results in greater microbial buildup, than the PVC material used in NFT channels. From these experiments, we recovered and classified 120 bacterial isolates belonging to 24 genera, with the most abundant being the genus Pseudomonas, followed by Bacillus. Unique groups of bacteria were recovered from biofilms from each PVC, pond liner and lettuce roots, as well as when comparing biofilm isolates to nutrient solution (planktonic) isolates. Objective 3: We first developed protocols for consistent infection of Pythium root rot, powdery mildew, and Xanthomonas sp. These protocols were then used to set up experiments to evaluate the effect of pathogen introduction in the resident microbial communities in lettuce roots and leaves when grown in DWC. In addition, experiments were run to determine the impact of potential biological control agents on Pythium disease in hydroponic lettuce production. Lastly, for experiments involving multiple microorganisms, we developed GFP-transformed Pseudomonas adapted to hydroponic systems. From these experiments, we determined that community structure differed in pathogen-inoculated plants compared with non-inoculated plants. For instance, when inoculated with Pythium, root samples with high disease severity presented higher bacterial and fungal ASVs richness compared to low disease severity and non-inoculated controls. Differential abundance analysis revealed that bacteria including Pseudomonas and Mucilaginibacter, and fungi such as Fusarium were enriched in inoculated plants. Whereas when leaf infection with powdery mildew was observed, only fungal communities were affected, most likely given the high severity of leaf infection observed. The team also evaluated the effect of pH-manipulation of nutrient solution and the application of sanitizers to the nutrient solution (SaniDate 12.0, 200 ppm) in eliminatingthe human pathogen Salmonella Typhimuriumfrom NFT hydroponic systems. The treatments tested (pH-modification or SaniDate) did not eliminate Salmonella. Further, Salmonella was transferred, or moved, from the nutrient solution to the leaves even after sanitizer application. The tested pH and sanitizer treatments did not impact the concentration of bioactive compounds in hydroponic lettuce, but the 200 ppm application of SaniDate in nutrient solution resulted in lower biomass than non-treated controls.
Publications
- Type:
Peer Reviewed Journal Articles
Status:
Under Review
Year Published:
2024
Citation:
Guevara F, Frey T, Malacrino A, Poeltra J, Benitez Ponce MS. Influence of system type and management practices on bacterial and fungal communities structure in hydroponic systems: Insights from Commercial Facilities in Ohio. Submitted to Phytobiomes Journal. Revised manuscript submitted. Available as preprint: agriRxiv.2024.00259, agriRxiv, doi:10.31220/agriRxiv.2024.00259.
- Type:
Peer Reviewed Journal Articles
Status:
Submitted
Year Published:
2024
Citation:
Vega,Isabella and Garay-Rodriguez,Gustavo and Lee,Ben and Moses,Campbell and Mock,Victoria and Almanza,Edward and Carni,Michael and Durocher,Shelley and Jacobs,Jonathan M. and Kaczmar,Nicholas S. and Mattson,Neil and Ponce,Maria Soledad Benitez and Heiden,Nathaniel. Submitted to HortTechnology. Available as preprint: agriRxiv.2024.00272, agriRxiv, doi:10.31220/agriRxiv.2024.00272
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2024
Citation:
Guevara F, Benitez Ponce MS. Insights into biocontrol potential of bacteria associated with hydroponic systems: phenotypic and whole genomic analysis. International Conference on Plant Pathogenic Bacteria & Biocontrol 2024.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2024
Citation:
Guevara F, Benitez Ponce MS. Pathogen-induced changes in bacterial and fungal communities structure in hydroponically grown lettuce. One Health Microbiome Symposium 2024.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2024
Citation:
Guevara F, Frey T., Benitez Ponce MS. Diversity and functional potential of microbial communities in leafy green hydroponic systems: Insights from commercial facilities in Ohio. Hayes Forum OSU, 2024.
- Type:
Theses/Dissertations
Status:
Published
Year Published:
2024
Citation:
Garay Rodriguez, G. A. (2024). Aeration, water source and surface material influence hydroponic lettuce production [Master's thesis, Ohio State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=osu1721036440171749
- Type:
Other
Status:
Other
Year Published:
2024
Citation:
Seminar: Benitez Ponce MS. Plant-microbiome networks impact plant productivity and mitigate plant disease and food safety risks in hydroponic production. USDA NIFA AFRI, Project Directors Meeting. Agricultural Microbiomes Program. August 22, 2024
- Type:
Other
Status:
Other
Year Published:
2024
Citation:
Seminar: Hart N. Examining the biocontrol Capabilities of bacteria against pythium. ORIP, OSU oral presentation. July 17, 2024.
- Type:
Other
Status:
Other
Year Published:
2024
Citation:
Seminar: Garay G. Aeration, water source, and surface materials influence hydroponic lettuce production. Deparment of Plant Pathology, The Ohio State University, Exit seminar. July 3 2024
- Type:
Other
Status:
Other
Year Published:
2024
Citation:
Seminar: Benitez Ponce. MS. Plant bacteriology and agricultural microbiomes lab. Department Seminar Spotlight. Deparment of Plant Pathology, The Ohio State University. February 5, 2024.
- Type:
Other
Status:
Other
Year Published:
2023
Citation:
Seminar: Guevara F. Exploring Microbial Communities in Leafy Greens Hydroponic Production Systems. Presented as a TALK at the Ohio Controlled Environment Agriculture Center (OHCEAC) annual meeting. Wooster, Ohio, USA. Dec 2023
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Progress 08/15/22 to 08/14/23
Outputs Target Audience:During the 2022-2023 period, we reached different audiences with this project, specifically leafy green hydroponic producers, and students and researchers in the areas of controlled environment agriculture, plant pathology, microbiology, and food safety. Specifically, during this period members of the team performed visits to four hydroponic leafy green facilities in the state. These visits were focused on sampling nutrient solutions, which were used as a starter material to seed greenhouse experiments focused on microbiome establishment. Our results have been shared with the research community through different venues, including invited seminars and webinars at various academic institutions and scientific societies, targeting audiences of undergraduate and graduate students, as well as faculty, other researchers, and producers. For example, Dr. Benitez Ponce was invited to apply to join the Phytobiomes Alliance and industry-academia collaborative effort in system-level research in agriculture. As part of this, Dr. Benitez Ponce presented a webinar, with international reach, and is currently the co-chair of Phytobiome Alliance's coordinating committee of the Controlled Environment Agriculture Working Group. We provided training in hydroponic production to a first-year undergraduate student researcher (OSU Agricultural Technological Institute, Plant Pathology major) and a visiting scholar, and continued the training of four graduate students (Plant Pathology and Human Nutrition), a postdoctoral researcher, and two research technicians. Finally, we are using material from our lettuce experiments as the source of root material for bacterial isolation and characterization in the Advanced Plant Pathology laboratory course (PP6002.01, Department of Plant Pathology at OSU). Changes/Problems:We continue to encounter delays in the shipment and availability of reagents and other consumables necessary for this research. We accommodate different reagents and consumables when it will not compromise the reproducibility of experiments. These also affect the turn-around time on services, including sequencing facilities. What opportunities for training and professional development has the project provided?Graduate student, Fiama Guevara attended and presented a poster at the 2023 MidWest Microbiome Symposium (May 2023), PI Soledad Benitez Ponce also attended the meeting. Several members of the team (Fiama Guevara, Christopher Taylor, Uttara Samarakoon) participated in the Ohio Controlled Environment Agriculture Center, annual meeting held July 2023. Graduate students, Fiama Guevara, Edwin Navarro, and Gustavo Garay attended and presented (G Garay oral, F Guevara, E Navarro, poster) at the 2023 Department of Plant Pathology Spring Symposium (May 2023). Graduate student Gustavo Garay attended and presented (oral) at the 2023 OSU Plant Science Symposium (Feb 2023), and will attend and present (poster) at the 2023 Plant Health Meeting (August 2023). Graduate student Abigail Mensa attended and presented at the 2023 meetings of the International Association of Food Protection and RKS 2023 How have the results been disseminated to communities of interest?The results have been disseminated to both the scientific community and hydroponic producers. As part of this, several seminars at academic institutions and scientific meetings, as well as poster presentations and short talks are listed under Products. In addition, our team continues to develop online and printed resources about hydroponic production and food safety management in leafy green hydroponics, which are available through the team's websites: https://u.osu.edu/benitezponce-1/links/ and https://producesafety.osu.edu/ What do you plan to do during the next reporting period to accomplish the goals?Activities to be completed in the next reporting period are indicated below by objective. Objective 1: Determine the differences in diversity and abundance of the resident microbiome of commercial lettuce hydroponic production with differing growing system setups. The team will continue with data analysis and manuscript preparation. Objective 2: Determine system management parameters that promote greater functional diversity of the resident microbiome of hydroponic lettuce production The team will generate and retrieve sequencing data associated with experiments of manipulation of nutrient solution (EC, pH, water source, and nutrient solution seeding). The data will then be analyzed and prepared for manuscript publication. We will complete the comparative genomic analysis of bacterial isolates. The whole genome sequencing information will then be correlated with phenotypic tests, as well as characteristics of the origin of each isolate. The data will then be prepared for manuscripts for publication. Objective 3: Evaluate hydroponic system resilience to plant and human pathogens, as a function of system management. Complete experiments with plant and human pathogens, and interactions with system management and resident members of the microbial community.
Impacts What was accomplished under these goals?
Accomplishments during the 2022-2023 are listed below by objective. Objective 1: Determine the differences in diversity and abundance of the resident microbiome of commercial lettuce hydroponic production with differing growing system setups. To characterize the diversity and relative abundance of bacteria and fungi associated with leafy green hydroponic production, we completed the library preparation, sequencing, and descriptive statistics of 736 samples obtained from ten commercial facilities in Ohio. Sample types included surface swabs, growing media, nutrient solution, lettuce leaves, and roots. Samples originated from facilities that varied in size (23-13150m2), system setup (Nutrient Film Technique (NFT), Deep Water Culture (DWC), Ebb & flow (EF), or Vertical Drip (VD)), a water source used for nutrient solution preparation, as well as pH and EC parameters of nutrient solution. We applied amplicon metabarcoding of the ribosomal markers, 16S rRNA, and ITS1 to survey bacteria and fungi, respectively. For roots, leaves, and nutrient solution, we recovered 16,963 and 3,533 taxa (or amplicon sequence variants (ASVs)) for bacteria and fungi, respectively. Community composition analysis indicated that the bacterial and fungal communities' structure differed between habitats (leaves, roots, and nutrient solution). Bacterial and fungal richness and evenness also showed differences between habitats, with roots and nutrient solution showing higher richness than leaves. However, bacterial evenness was significantly higher in leaves, whereas fungal evenness was higher in leaves and nutrient solution, compared to roots. Further comparison of root samples between production systems showed higher bacterial richness, but lower evenness in DWC systems, whereas greater fungal richness and evenness were observed in EF and NFT systems. Lastly, communities from nutrient solution were less variable across facilities, compared to other sample types. The most prevalent bacterial genus in root samples was Flavobacterium; whereas in leaves, an unclassified Comamonadaceae and Anhiella were prevalent. Additionally, fungal genera Olpidium and Golovinomyces were prevalent in both leaves and roots. And, consistent groups of bacteria were recovered from the nutrient solution (through amplicon metabarcoding and culturing), including the genera Bosea, Sphingomonas, and Sphingobium. Objective 2: Determine system management parameters that promote greater functional diversity of the resident microbiome of hydroponic lettuce production Our research focused on greenhouse experiments manipulation of two characteristics, nutrient solution management, and system setup (DWC vs NFT and associated surface materials). For the nutrient solution management experiments, we manipulated nutrient content, through electrical conductivity (EC), pH, and the water source. Experiments for EC and pH manipulation were completed in both experimental DWC and NFT; whereas water source experiments were completed in a small-scale, 5-gallon DWC setup. As indicated in the literature, greater lettuce vigor was observed under recommended conditions of 1.8 EC and pH of 5.8; however, we recovered more culturable bacteria (colony-forming units) from roots of lettuce grown at an EC of 2.5. From our pH experiments, we determined that maintaining a consistently low pH (5.0) in a production system is challenging and is related to changes in the nutrient solution as the lettuce head begins to form. This resulted in inconclusive data on pH effects on bacterial abundance in roots. Lastly, lettuce vigor, when grown in a nutrient solution prepared with rainwater (RW), was higher, than when grown in a nutrient solution prepared with reverse osmosis-filtered municipal water. In addition, a subset of roots from lettuce grown in RW nutrient solution harbored diverse bacterial communities, which differed in dominant bacterial taxa, compared to municipal water. Plant and growth media samples from these experiments (nutrient solution management) have been processed for nucleic acid extraction and microbial communities will be further characterized through amplicon metabarcoding. In addition to testing parameters of nutrient solution characteristics, we also tested the effect of spent, or reused nutrient solution, and its community composition, on subsequent lettuce growth cycles. This experiment allowed us to determine if: a) nutrient solution can be used to seed, or inoculate, a leafy green hydroponic setup and b) the degree of contribution of the past nutrient solution, compared to the current setup, on hydroponic system microbial community composition. From two experimental runs, which included seeding with nutrient solution from active, commercial hydroponic facilities, no significant effect on lettuce biomass was observed, regardless of nutrient solution origin and the richness of bacteria recovered from the nutrient solution was lower at the end of the experiment, compared to the original "seed". Analysis of the effect of nutrient solution seed in lettuce root microbial communities is ongoing. For system type (DWC vs NFT) characteristics, we focused on the impact of surface materials (i.e. polyethylene liner and PVC, for DWC and NFT, respectively) on the microbial buildup of biofilms and the diversity of bacteria from surface swabs. Our results indicated that even though the DWC liner results in greater microbial buildup, the diversity of bacteria recovered was greater from PVC channels. In addition, the surfaces within an NFT channel accumulated different amounts of bacteria, than the surface of the nutrient tank, and the nutrient solution. To evaluate functional diversity in the microbial communities of hydroponic system production, we are characterizing the activity and genome composition of isolates recovered from nutrient solution and surface biofilms. A total of 287 isolates have been classified into 67 genera, with the most abundant being the genus Pseudomonas. Other dominant genera differed between samples, with Bacillus being predominant in surface samples. A subset of isolates produced siderophores, can solubilize phosphorous, inhibit pathogen growth in vitro and in plants, and show the potential to fix nitrogen. 72 of these isolates have been selected for whole genome sequencing. Objective 3: Evaluate hydroponic system resilience to plant and human pathogens, as a function of system management. For this objective, in the current reporting period, we first tested the efficacy of inoculation protocols for the application of the leaf and root pathogens in hydroponically grown lettuce (deep water cultures). As a result of this, we developed protocols that result in consistent infection of Pythium root rot, powdery mildew, and Xanthomonas sp. These protocols were then used to set up experiments to evaluate the effect of pathogen introduction in the resident microbial communities in lettuce roots and leaves when grown in DWC. In addition, experiments were run to determine the impact of potential biological control agents on Pythium disease in hydroponic lettuce production. Lastly, for experiments involving multiple microorganisms, we currently have available GFP-transformed isolates adapted to hydroponic systems. The team also evaluated the effect of pH-manipulation of nutrient solution and the application of sanitizers to the nutrient solution (SaniDate 12.0, 200 ppm) in eliminatingthe human pathogen Salmonella Typhimuriumfrom NFT hydroponic systems. The treatments tested (pH-modification or SaniDate) did not eliminate Salmonella. Further, Salmonella was transferred, or moved, from the nutrient solution to the leaves even after sanitizer application. The tested pH and sanitizer treatments did not impact the concentration of bioactive compounds in hydroponic lettuce, but the 200 ppm application of SaniDate in nutrient solution resulted in lower biomass than non-treated controls.
Publications
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2022
Citation:
Benitez Ponce MS, Guevara F, Frey T, Taylor L. System design, surface characteristics, and recirculation influence microbial communities in hydroponic leafy green production. International Phytobiomes Conference, 2022. Abstract. Accepted for presentation but presentation canceled due to illness.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2023
Citation:
Benitez Ponce MS, Guevara F, Frey T, Garay G, Taylor L. Plant and Animal Genome Meeting. January 2023. Exploring Phytobiomes Workshop speaker. Title: System Design, Surface Characteristics, and Recirculation Influence Microbial Communities in Hydroponic Leafy Green Production.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2023
Citation:
Garay G, Benitez Ponce MS. Impact of aeration and water source on yield and bacterial abundance in deep water culture hydroponics. Plant Science Symposium, The Ohio State University. February 2023
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2023
Citation:
Mensah, AA., Melanie. ILL., TM., Ilic, S. (2023) Mitigation of Salmonella Typhimurium in Nutrient Film Technique (NFT) hydroponic system for improved food safety and nutrition.RKS 2023
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2023
Citation:
Rivas, G., Mensah, AA., Collin B., Melanie. ILL., Ilic, S. (2023) Food Safety of Hydroponic Fresh Produce: A Scoping Review, IAFP 2023
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2023
Citation:
Mensah, AA., Melanie. ILL., TM., Ilic, S. (2023) Eliminating Salmonella Typhimurium from lettuce grown in NFT hydroponic system for improved food safety and nutrition, IAFP 2023
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2023
Citation:
Guevara F, Navarro E, Benitez Ponce, MS. Functional characterization of Bacteria associated with lettuce hydroponic production systems in Ohio. Midwest Microbiome Symposium. May 2023.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2023
Citation:
Garay G. Comparison of different water sources for hydroponic lettuce production. Department of Plant Pathology, The Ohio State University, Spring Symposium. May 2023.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2023
Citation:
Guevara F, Frey T, Benitez Ponce, MS. Unraveling microbial communities of commercial lettuce hydroponic production systems in Ohio. 4th Plant Microbiome Symposium, Quito-Ecuador, 2023. Abstract. Accepted for presentation but presentation canceled due to travel cancelations.
- Type:
Conference Papers and Presentations
Status:
Awaiting Publication
Year Published:
2023
Citation:
Garay G, Benitez Ponce MS. 2023. Water Source influence on lettuce on controlled environment system. Plant Health 2023.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2023
Citation:
Navarro, E. Uncovering T6SSs in a collection of Pseudomonas spp. Department of Plant Pathology, The Ohio State University, Spring Symposium, poster presentation. May 2023.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2023
Citation:
Guevara-Guillen F. Bacterial communities in lettuce hydroponic systems. Department of Plant Pathology, The Ohio State University, Spring Symposium, poster presentation. May 2023.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2023
Citation:
Benitez Ponce MS. Drivers of Microbial Community Composition in Hydroponic Leafy Green Production. Phytobiomes Alliance, Webinar Series. May 24, 2023. https://youtu.be/uNCAOyxsN2A
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2022
Citation:
Benitez Ponce MS. Spatial distribution of microbial communities in agricultural systems.
Michigan State University, Department of Plant, Soil and Microbial Sciences. Invited Seminar Speaker. Oct 12, 2022.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2023
Citation:
Navarro E. El uso de la microbiolog�a en la producci�n agr�cola: �nfasis en el uso de Pseudomonas como control biol�gico Sociedad Estudiantil de Microbiolog�a Industrial- Universidad de Puerto Rico, Mayag�ez.February 9, 2023
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Progress 08/15/21 to 08/14/22
Outputs Target Audience:During the 2021-2022 period we reached different audiences with this project. Our efforts were targeted toward leafy green hydroponic producers in the state of Ohio, and students and researchers that work in different aspects of plant production, controlled environment agriculture, plant pathology, microbiology, and food safety. Specifically, in 2021-2022 we performed two visits to leafy green growers in Ohio state to sample from their hydroponic facilities andassess microbes in their production system. This amounts to a total of 10 leafy green hydroponic facilities sampled in this project.Additional visits were performed to support individual growers' needs, in particular, to discuss specific questions on food safety in leafy-green hydroponic production. As part of the efforts on food safety, updates on good agriculture practices to promote food safety in hydroponic productions were published, and an online self-paced course was developed for producers. We also provided training in hydroponic production to three undergraduate student researchers, and continued the training of two graduate students, two postdoctoral researchers, and a research technician. Our results have been shared with the research community through different venues, including invited seminars at various academic institutions and scientific conferences. Finally, we are incorporating small experiments in our hydroponic setup into hands-on laboratory courses (Department of Plant Pathology at OSU), focused on plant-microbial interactions. Changes/Problems:During this reporting period, we encountered delays due to the availability of reagents and other consumables necessary for nucleic acid extractions. To solve this challenge, we had to modify protocols to use materials more readily available, but this has resulted in delays in the sequencing process. What opportunities for training and professional development has the project provided?Graduate student, Fiama Guevara attended the in person meeting of the American Phytopathological Society in August 2022. Several members of the teamparticipated in different roles at the Ohio Controlled Environment Agriculture Center, annual meeting held July 2022. Among the organizers of the 2022 meeting, Dr. Christopher Taylor is a co-PI on this project. The topic of this annual meeting was "Advancement of Microbial Technologies for Controlled Environment Agriculture", and we had two presentations by members of our team (Frey et al and Ilic et al). In addition, undergraduate students, graduate students, postdocs and researchers from multiple labs attended this meeting. Graduate student, Fiama Guevara attended the Microbiome Informatics Workshop, organized for OSU Centers of Microbiome Science, Fall 2021. Postdoctoral researcher Timothy Frey, graduate student Fiama Guevara, and undergraduate student Gustavo Garay participated in OSU's Code Club during the period Fall 2021-Summer 2022. How have the results been disseminated to communities of interest?The results have been disseminated to both the scientific community and hydroponic producers. As part of this, several seminars at academic institutions and scientific meetings, as well as poster presentations and short talks are listed under Products. In addition, our team prepared printed material, online resources, and performed grower visits to contribute to food safety management in leafy green hydroponics. What do you plan to do during the next reporting period to accomplish the goals?Activities to be completed in the next reporting period are shown below by objective. Objective 1: Determine the differences in diversity and abundance of the resident microbiome of commercial lettuce hydroponic production with differing growing system setup. Retrieve the sequence data from samples of all commercial facilities. Complete the data analysis and publication of the microbial community characterization from the commercial hydroponic facilities visited. Prepare data to share with individual growers Objective 2: Determine system management parameters that promote greater functional diversity of the resident microbiome of hydroponic lettuce production. Repeat experiments on the influence of pH of the nutrient solution on lettuce health and associated microbial communities in DWC and NFT. Complete experiments on the water source as a determinant of microbial community composition in hydroponic lettuce roots. Setup, run experiments, and evaluate the effect of biological inoculants on microbial community composition in hydroponic lettuce. Perform comparative genomic analysis of bacterial isolates obtained from diverse types of hydroponic systems. Analyze data and prepare manuscripts for publication. Objective 3: Evaluate hydroponic system resilience to plant and human pathogens, as a system management function. Setup and run experiments to evaluate hydroponic system response, including microbial community dynamics, to inoculation with individual plant or human pathogens, or a combination thereof. Analyze data and prepare manuscripts and other materials toshare our results with stakeholders.
Impacts What was accomplished under these goals?
Accomplishments during 2021-2022 are listed below by objective. Objective 1: Determine the differences in diversity and abundance of the resident microbiome of commercial lettuce hydroponic production with differing growing system setups. Established contact, visited, and sampled two additional commercial leafy green hydroponic facilities in Ohio. This sums to a total of 10 commercial facilities visited (and sampled) throughout the duration of this project. One facility, out of the 10, was selected to perform multiple samplings during this project. During 2021-2022 we completed the second sampling visit. Completed nucleic acid extraction from plant material, nutrient solution, and system surface samples obtained from all commercial facilities and time points sampled. Submitted nucleic acids for amplicon sequencing of bacterial and fungal ribosomal gene markers and sequencing is currently in progress. Objective 2: Determine system management parameters that promote greater functional diversity of the resident microbiome of hydroponic lettuce production. Completed two experiments on the influence of nutrient solution electrical conductivity on lettuce growth and associated microbial communities in research DWC. Completed an experiment on the influence of pH of nutrient solution on lettuce health and associated microbial communities in each DWC and NFT systems. Designed and set up experiments on water source influence in lettuce roots and associated microbial communities in DWC. Completed experiments to evaluate different types of sanitizers on NFT surfaces. Determined the effects of system surface characteristics (PVC for NFT and pool liner for DWC) on bacterial biofilm formation and composition. Performed experiments to evaluate differences in bacterial biofilm formation and composition concerning location in a leafy green hydroponic system (i.e. water tank, proximal to the Rockwool plug, root tip, end of the NFT channel). Objective 3: Evaluate hydroponic system resilience to plant and human pathogens, as a function of system management. Tested inoculation methods for consistent Pythium infection in lettuce DWC. Tested for the survival of human pathogens in leafy green NFT and DWC.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2022
Citation:
Malacrino A; Abdelfattah A; Berg G; Benitez MS; Bennett AE; B�ttner L; Xu S; Schena L. 2022. Exploring microbiomes for plant disease management. Biological Control. 169: 104890. doi.org/10.1016/j.biocontrol.2022.104890.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2022
Citation:
Guevara F, Frey T, Taylor L, Benitez Ponce MS. Isolation and identification of bacteria associated with Nutrient Film Technique (NFT) hydroponic production systems in Ohio. Plant Health 2022.
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2021
Citation:
Benitez Ponce MS. V International Phytopathology Symposium. Universidad San Francisco, Quito-Ecuador. Sep 15, 2021. Keynote presentation. Title: Din�mica de comunidades microbianas en ecosistemas agr�colas.
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2022
Citation:
Guevara F. Understanding the role of microbial communities in lettuce hydroponic production systems. Department of Plant Pathology, OSU, Spring Symposium. May 9, 2022.
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2022
Citation:
Frey T, Guevara F, Taylor L, Benitez Ponce S. Microbial communities and biofilms associated with hydroponic lettuce production systems in Ohio. Ohio Controlled Environment Agriculture Center Conference. July 20, 2022.
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2021
Citation:
Benitez Ponce MS. Millersville University, Department of Biology Colloquim Speaker. Oct 20, 2021. Title: Microbial community dynamics in agricultural systems.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2022
Citation:
Mensah, Abigail Aba, et al. "Mitigation of Human Pathogen Contamination in Lettuce and Basil Produced in NFT Hydroponic System." Current Developments in Nutrition 6.Supplement_1 (2022): 522-522.
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2022
Citation:
Ilic S. Human pathogens in plant production systems. Master in Plant Health Management Program, Seminar Series, The Ohio State University, Department of Plant Pathology. March 28, 2022.
- Type:
Conference Papers and Presentations
Status:
Other
Year Published:
2022
Citation:
Ilic S, Ivey M. Survival of Foodborne Bacterial Pathogens in Leafy Green Hydroponic Production Systems. Ohio Controlled Environment Agriculture Center Conference. July 20, 2022.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2022
Citation:
Sirsat S, Dong M, Ilic S. Mixed Methods Approaches to Investigating Microbial Produce Safety Hazards and Mitigation in Hydroponic and Aquaponic Operations. International Association of Food Protection. August 2022.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2022
Citation:
Mensah, AA, Ivey ML, Miller TM, Ilic S. Impact of Sanitizers on Nutrient Film Technique (NFT) Grown Lettuce and Basil. International Association of Food Protection. August 2022.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2022
Citation:
Moodispaw MR, Ivey ML, Ilic S. Effectiveness of Surface Sanitizers Against Salmonella Typhimurium in Hydroponic Lettuce System. International Association of Food Protection. August 2022.
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Progress 08/15/20 to 08/14/21
Outputs Target Audience:Our efforts were targeted to reach leafy green hydroponic producers in the state of Ohio. We contacted, visited, and/or communicated with 9 production facilities. The facilities ranged in size, number of heads produced, and system design. In addition, we worked with graduate students, research technicians, and postdoctoral researchers. Changes/Problems:The greatest challenge we encountered during this reporting period was the difficulty to visit and have access to production facilities due to the COVID-19 pandemic. In addition, the pandemic resulted in delays in recruiting both graduate students and postdoctoral researchers. The team is currently complete and many of the restrictions to visit facilities have been lifted. What opportunities for training and professional development has the project provided?As part of our project, and to train project participants (post-doctoral researcher, research technician, and graduate students) we developed a 6-hour workshop on experimental design considerations and data analysis of amplicon sequencing experiments for plant microbiome analysis. This workshop was imparted to a total of 12 participants. How have the results been disseminated to communities of interest?We have had direct communication with hydroponic producers. The goal of this has been to inform our project objectives and engage a greater number of facilities to participate. In addition, the scope and goals of this project, as well as preliminary was shared with the scientific community through a poster presentation at the Virtual Conference of the American Phytopathological Society (Plant Health 2021) and as part of a virtua seminar series at the University of California, Santa Barbara. What do you plan to do during the next reporting period to accomplish the goals?During the next reporting period we will complete the sampling, sequencing, and data analysis of the microbial community surveys of the facilities we have identified, lined up and visited during the first year of this project. In addition, experiments will be established in the greenhouse to test specific management conditions that influence microbiota and plant health in DWC and NFT. Conditions to be managed include source or water, nutrient solution characteristics (pH, EC), sanitation strategies (UV, bleach), and biological inoculants (for plant growth promotion/biological control). For these experiments, microbial communities will be assessed using different methods, including culturing, amplicon sequencing, and metagenomic approaches.
Impacts What was accomplished under these goals?
Microbial communities of plants grown in hydroponic systems have not been thoroughly studied in contrast with the efforts in soils. In hydroponics, plant roots are suspended in a nutrient solution, and the complexity of the ecosystem is lower. Two production systems are most common for leafy greens, Nutrient Film Technique (NFT) and Deep Water Culture (DWC). These two systems differ in the delivery mode of nutrients. In NFT roots grow in a thin layer of a nutrient solution which flows at a constant rate through PVC channels. Whereas in DWC, roots are completely immersed in a pool of aerated nutrient solution. In our preliminary studies, we showed that the diversity of microorganisms associated with lettuce roots and nutrient solution is related to the hydroponic system setup. In this project we aim to characterize the microbial communities in commercial hydroponic lettuce production; and, taking advantage of the environmental control systems, use this information to develop models of microbial community management and contributions to plant health. We will further manipulate the conditions of the nutrient solution in our research greenhouses and test the effect of the resident hydroponic microbiota on plant pathogen and human pathogen spread and establishment. We have established collaborations with nine commercial leafy green production facilities across Ohio. These facilities range in size (from 23 to 13150 m2) and production capacity (up to a million heads per year). Based on the characteristics of the production facilities, we have identified a common set of samples to be collected, which will allow for comparisons across locations. The sampling design targets plant tissue (shoot and root), nutrient solution, and water sources, as well as operational surfaces. We have worked on the standardization and reproducibility of techniques used for sampling, nucleic acid extraction, and recovery of microorganisms from water, nutrient solution, system surfaces, and other facility zones. Our sampling strategy also incorporates environmental sampling zones to monitor members of the hydroponic microbiome that could represent a potential food safety concern. In addition, in collaboration with one of our production facilities, we designed a DWC system to be used for experimentation in our research greenhouses. We currently have the capacity to set up replicated experiments with nine independent DWC, each holding 56 lettuce heads. Together with the already available NFT systems on campus, we plan to manipulate growing conditions to compare and contrast microbiome and their manipulation across hydroponic systems.
Publications
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2021
Citation:
F. E. Guevara, T. Frey, A. Malacrin�, Maria-Soledad Benitez. 2021. Understanding microbial communities composition for management practice improvement in lettuce hydroponic systems in Ohio. APS Plant Health. Virtual poster presentation. August 2-5 2021.
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