Source: SYNERGISTIC HAWAII AGRICULTURE COUNCIL submitted to
COORDINATED APPROACH TO COFFEE LEAF RUST
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
NEW
Funding Source
Reporting Frequency
Annual
Accession No.
1027511
Grant No.
2021-51181-35900
Project No.
HAWW-2021-07760
Proposal No.
2021-07760
Multistate No.
(N/A)
Program Code
SCRI
Project Start Date
Sep 1, 2021
Project End Date
Aug 31, 2025
Grant Year
2021
Project Director
Shriner, S.
Recipient Organization
SYNERGISTIC HAWAII AGRICULTURE COUNCIL
190 KEAWE ST STE 25
HILO,HI 967202849
Performing Department
(N/A)
Non Technical Summary
The October 2020 discovery of Coffee Leaf Rust (CLR), Hemileia vastatrix, on both Maui and Hawaii Islands brought the most devastating coffee disease in the world to the state. Present in Puerto Rico since 1989, the fungus attacks the leaves of coffee trees causing severe defoliation, yield losses of 70% or greater, and possible tree death. The disease is especially challenging to control, and is a significant threat to the 1,470 growers in Hawaii and the 2,300 Puerto Rican growers. Coffee is ranked as a top commodity in in both regions, but individual farms are small and classify as socially disadvantaged. In Kona alone, there are hundreds of coffee farms that contribute not only to the economic viability of Hawaii, but to the preservation of a cultural heritage dating back 150 years. Hawaii coffee has downstream benefits of value-added roasting, farm jobs, ecotourism, and ancillary industries calculated at over $500 million annually. Data for Puerto Rico is not reliable post-Hurricane Maria, but similar to Hawaii, the majority of growers are socially disadvantaged growers, and provide a strong economic benefit to their communities.Research from this grant will provide quantifiable benefits to the US coffee industry and beyond, including a wide range of agronomic, economic, and genomic assistance. Benefits include field trials of new rust-resistant coffee tree varieties that meet the exacting standards of the high-value Hawaii industry and may increase the quality of coffee originating from Puerto Rico, thus raising the standard of living for growers.Until the new varieties are ready for propagation, field research and extension will provide immediate solutions to maintain the tree health of current, susceptible varieties.Economic analyses will provide a deep look at US-grown coffee from both a growing and a selling perspective; this evaluation has not been done in the modern specialty coffee era. Finally, analysis of CLR and coffee genomics will provide a closer look at a pathogen that is not yet understood on a genetic basis and identify the host/pathogen interaction that could lead to a better understanding of how the disease spreads and methods for how growers can control the fungus in their fields.
Animal Health Component
0%
Research Effort Categories
Basic
70%
Applied
20%
Developmental
10%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
2162232106065%
2012232108015%
6012232301010%
2152232110210%
Keywords
Goals / Objectives
Objective 1: Plant Propagation of CLR-Resistant VarietiesCoffee leaf rust resistant coffee cultivars offer a sustainable long-term solution to combating Coffee Leaf Rust in Hawaii and Puerto Rico. However, both markets need to maintain high "cupping" quality standards to satisfy their specialty coffee market. In addition, due to the high input and labor costs in Hawaii, easily managed and high-yielding cultivars are needed to provide lower production costs. Identifying the resistant cultivars and determining the specific CLR race in HI and PR will drive the long-term strategy. However, this resistance is often associated with increased bitterness. Careful evaluation is needed to ensure high cup quality for the specialty coffee market.Both regions will join the global World Coffee Research varietal trials, and identify a suite of potentially suitable varieties. Once identified, scientists, growers, and industry members will jointly determine which CLR resistant coffee varieties can be imported, trialed for cup quality and agronomic performance under local conditions, and then propagated. Harvest yield can be drastically reduced in varieties susceptible to losses due to nematodes; CLR-resistant cultivars will also be evaluated for resistance to the Kona coffee root-knot nematode (Meloidogyne konaensis). Vigilance against introducing new CLR races to HI and PR is also essential; thus, rapid molecular methods to detect and identify new CLR introductions are essential.Goal 1A: Field Trials of CLR Resistant VarietiesResearchers will conduct side by side comparisons, in typical field conditions, to assess yields, quality, general plant characteristics, and plant health including susceptibility to CLR and other diseases. Extension will facilitate meetings with industry members to assist with cup quality determinations.Goal 1B: Propagation of CLR Resistant VarietiesHawaii currently has limited capacity to produce planting material of putative CLR resistant varieties at commercial scale. Micropropagation through somatic embryogenesis (SE) is a powerful tool to rapidly produce large quantities of plants from a limited initial supply. The methods for SE are well developed for coffee. This project will move in three stages: Stage 1, Establishing embryo culture in preparation for large scale micropropagation; Stage 2, Setup of Bioreactor Facility; and Stage 3: Plantlet production and acclimatization.Goal 1C: Root Knot Nematode (RKN) compatibility trialsThe RKN, Meloidogyne konaensis, is widespread in coffee regions and responsible for significant yield declines. New CLR resistant varieties will be inoculated in potted assays to determine resistance to RKN and whether they will need to be grafted or can be planted directly into potentially infested soil. Resistant varieties will be confirmed through field tests to evaluate both rootstock-scion compatibility as well as field performance.Objective 2: CLR Survey and Monitoring ProgramSurveys for CLR incidence and severity will allow the spread of the disease to be tracked across the coffee-growing landscape, and in conjunction with management records this information will be used to develop IPM guidelines for CLR. The environmental factors that are correlated with the disease must be determined to better understand any regional and/or seasonal differences in spore germination and survival. Information on incidence, severity, pathogen movement and weather variables will be used to parameterize and build predictive models so that resources can be targeted to locations forecasted to be high-risk, thereby mitigating impacts to production before symptoms appear. Data collected through surveys and monitoring will also inform the selection of rust-resistant varieties in each region and guide the development of Extension recommendations for the timing of fungicide applications. Extension staff will work closely with researchers and SHAC to relay results to growers promptly.Objective 3: CLR Field Management Research ProgramThe development and availability of rust resistant varieties will encompass a multi-year timeline for the industry to transition to new cultivars. In the meantime, immediate responses are needed to control CLR in the field.Objective 3A: Field and laboratory research to identify effective and safe fungicides (systemic, contact, and/or biological control products)Fungicides will be an important management tool for coffee leaf rust, but copper fungicides must be present on the leaf prior to infection, and copper soil accumulation is a major environmental concern. Curative treatments include the use of systemic fungicides (triazoles and strobilurins), but most systemic fungicides are not registered for use on coffee in Hawaii. Therefore, identifying fungicides and/or biological control products that are effective against CLR in Hawaii and Puerto Rico is critical to sustain productive coffee orchards. We will determine the appropriate timing for rotation of products to avoid developing fungicide resistance or copper toxicity in the soil. Special Local Need Registration and/or other appropriate venues to facilitate the use of effective agents identified, if any, will be pursued to use on coffee. Viable control methods will be rapidly deployed through Extension activities.Objective 3B: Biological Control of CLRThe goal of this objective is to survey, isolate, identify, and evaluate local strains of CLR mycoparasites and herbivores. We will determine their potential to destroy the rust's reproductive structures and reduce CLR incidence and severity in the field. In addition to assays for effectiveness, taxonomic studies may be necessary in order to determine host specificity, a necessary step for potential biocontrol. Studies of growth optima, such as temperature, niche and seasonality may also be necessary to identify key determinants for optimal deployment in different environments.Objective 3C. Optimizing Field Health for CLR Management in Non-Resistant VarietalsField management data for non-resistant varietals are necessary for development of Good Agricultural Practices and Extension use. We will examine various methods of pruning, sanitation practices, fertilizer regimes, planting density, soil fertility, and plant nutrition to optimize field health in non-resistant coffee varietals.Objective 4: Pathogen and Host CharacterizationGoal 4A: Genetics of CLRCLR races present in HI and PR are unknown, but race typing is essential for optimal deployment of resistant cultivars in the field, and genotyping using these markers could be a more efficient way to do this. Scientists from ARS and Purdue University will develop a variable marker set that is reliable, repeatable, and capable of distinguishing CLR isolates in Hawaii and Puerto Rico from others around the world.Goal 4B: Understanding Resistance to CLRThe mechanism that confers CLR resistance to a varietal is not yet known. A similar RNAseq project on susceptibility of Coffee Berry Borer to the fungus B. bassiana demonstrated an ability to use the methodology to consider and analyze such data.Objective 5: Economic Analysis and ImpactsEach aspect of the project will include an economic analysis to better understand the costs, benefits, and tradeoffs across different control strategies, both from the perspective of coffee growers and the larger coffee marketplace. It will also include analyses for replanting with resistant varieties versus costs to maintain control with current (non-resistant) varieties under IPM protocols.
Project Methods
Objective 1: Plant Propagation of CLR-Resistant VarietiesObjective 1A: Field Trials of CLR Resistant VarietiesSeedlings will be planted following World Coffee Research (WCR) protocols on commercial farms in Hawaii and at research facilities in Puerto Rico. Planting will follow a Fisher block design with three replicated blocks with ten plants per accession randomized across each block. Each planting of imported coffee varieties will also include a CLR susceptible industry-standard cultivar and a CLR resistant cultivar.Resistance to both natural and inoculated CLR infection will be monitored. Management practices will be recorded for each variety trial including pruning, fertilization, fungicide, and pesticide applications as well as soil and leaf tissue analyses. Nutrient data, timing of fertilizer application, and CLR incidence and severity will be compared to literature to determine fertilizer recommendations. In addition, pruning will be evaluated, including time of stump, shoot removal, leaf litter removal, and the number of leads.Data collected will include: vegetative growth, CLR incidence and severity, other pest and disease incidence, flowering, cherry phenology, production data (date and weights of harvest), and bean quality (weight, size, imperfections, and grade).To finalize recommendations, extensive quality assessments on flavor will be completed by growers, processors and coffee professionals. Extension staff will participate in panels and disseminate results to all growers to better facilitate decision-making on varietal choices.Objective 1B: Propagation of CLR Resistant VarietiesSomatic embryogenesis is a plant tissue culture method used to clonally multiply unique plant lines, and very large numbers of plants can be produced. This effort will move forward in three stages: Stage 1, Establishing embryo culture in preparation for large scale micropropagation; Stage 2, Set up of Bioreactor Facility; and Stage 3: Plant production and acclimatization.Objective 1C: Root-knot Nematode Resistance Trials and Enhancement of Resistance ScreeningCLR-resistant coffee cultivars obtained from WCR trials and other sources will be screened against M. konaensis in a greenhouse potted plant bioassay. This assay will determine resistance to this pest and whether cultivars will need to be grafted on nematode-tolerant rootstocks. Long term field studies of promising cultivars will be initiated in M. konanesis-infested fields. Also, faster screening mechanisms will be developed. Objective 2: CLR Surveys and Environmental MonitoringMonthly incidence and severity surveys will occur on 17 commercial coffee farms on Hawaii Island and up to 20 sites in Puerto Rico. In addition, two abandoned coffee sites will be used to track the epidemiology of CLR in unmanaged environments. During each evaluation, 25 coffee trees will be randomly sampled (number of leaves examined per survey = 700-1000). To estimate disease severity for each site/sampling date, one infected leaf per tree (25 infected leaves) will be scored for percent infected area using ImageJ. Data collected will be subjected to coefficient of variation analyses to optimize this sampling strategy and determine the minimum number of samples required to attain accurate estimates.The area under disease progression curve (AUDPC) will be estimated as a quantitative summary of disease intensity over time using the Agricolae package in R (R Core Team 2020). Measurements taken under constant environmental conditions in the laboratory will be combined with experimental and observational data from field surveys to help describe the CLR life cycle stages and correlate symptom development and microclimate variables for optimized management including timing of fungicide sprays.Active and/or passive spore samplers will be installed to characterize airborne pathogen movement. For environmental monitoring, ARS will use equipment it has in place: RX3000 remote cell-service weather equipped with sensors to measure air and soil temperature, relative humidity, rainfall, wind speed and direction, solar radiation, soil moisture, and leaf wetness. Correlation analyses will be conducted between disease incidence and severity and particular microclimate variables for development of models.Objective 3: CLR Field ManagementObjective 3A: Field and laboratory research to identify effective and safe fungicides (systemic, contact, and/or biological control products)Fungicidal and biocontrol activity will be tested in the lab and on coffee plants in greenhouse and field assays. The incidence and severity of CLR and the area under the disease progression curve (AUDPC) will be evaluated at 10-12-day intervals according to diagrammatic scales.Efficacy field research trials will be conducted in plots located on collaborating private coffee farms. Research trials will consist of 5 treatments including 4 systemic fungicides and one commercially available biological control product, plus an untreated control. Treatments will be selected because they are in use to control CLR in other countries (but not US) and do not overlap with ongoing IR-4 trials. Foliage of each tree will be evaluated for incidence, severity and possible crop injury.A Special Local Need Registration will be pursued for any successful products identified. Extension will be included in all discussion of results and will update publications as products become available to growers.Objective 3B: Biological Control of CLRField surveys will be conducted at farms varying in elevation and microclimate in multiple coffee-growing regions to determine at what times of year and under what conditions mycoparasitic infection of CLR is most common. Leaf samples with symptoms of rust and signs of possible mycoparasites will be collected from trees and leaf litter. During surveying and monitoring (Objective 2), mycoparasite frequency will be estimated to determine how widespread the infections are in the current CLR populations on each farm to understand the conditions that favor its growth. Field and lab tests will determine if local Lecanicillium-type fungi are effective at controlling CLR, what mix of species is most effective, and what conditions and seasons favor the infection. Field, lab and DNA data will be integrated to develop potential biocontrol agents and techniques. Mixtures of species and formulations will be tested to optimize infection of CLR.Objective 3C. Optimizing Field Health for CLR Management in Non-Resistant VarietalsFollowing the protocols established in Objective 1A, management practices will be recorded for non-resistant varietals. As research results become available, the dissemination of these results and recommendations for producers will be communicated by Extension.Objective 4: Pathogen and Host CharacterizationObjective 4A. Genetics of CLRThe approach to understanding and utilizing the genetics of CLR includes five parts: 1) Genome sequencing and assembly of H. vastatrix; 2) Genome mining for sensitive markers for population genetics; 3) Population genetics analysis and phylogeography of HI and PR isolates; 4) Race characterization of HI and PR isolates (in conjunction with Obj. 1A); and 5) GWAS analysis of genomes to find genes associated with virulence.Objective 4B: Understanding Resistance to CLRResearch will compare CLR resistance in two susceptible and two resistant cultivars from Objective 1A. One race of CLR, tested by inoculation on differential plantswill be used. RNA will be extracted from leaves and RNAseq will be used to seek for proteins and genes that are differentially expressed between susceptible vs. resistant plants during infection.Objective 5: Economic Analysis and ImpactsEach aspect of the project will include an economic analysis to better understand the costs, benefits, and tradeoffs across different control strategies, both from the perspective of coffee growers and the larger coffee marketplace.

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

Outputs
Target Audience:A key audience target for Objectives 1, 2, 3, and 5 are coffee growers and extension agents.Data collected under Objective 1 will benefit the coffee industry in Hawaii and Puerto Rico by identifying high-quality coffee varieties that are resistant to CLR and other diseases for long-term replanting of damaged fields.In the immediate term, recommendations generated by Objectives 2 and 3 will assist growers with agronomic field management on susceptible trees.Economic analyses under Objective 5 will inform growers about the cost effectiveness of each recommendation.Specific outreach and extension efforts are described further under the Accomplishments: Communities of Interest section. Information sharing between Hawaii and Puerto Rico scientists will benefit the national and international scientific community.The genomic activities under Objective 4 are targeted to researchers, plant breeders and plant pathologists. Knowledge generated under this grant strengthens the coffee industry in Historically Underserved and economically depressed areas like the mountain region of Puerto Rico and the Ka'u region of Hawaii by maintaining and creating jobs in farms and coffee processing organizations. These two regions are denoted on the Federal government'sJustice40 Initiative map. Changes/Problems:A significant delay has arisen due to the Hawaii Department of Agriculture's (HDOA)Plant Quarantine division. Shifting protocols and loss of staff within the Department has delayed permitting under Objective 1. Each new cultivar requires a 1 year quarantine, but new protocols for completingquarantineare currently under development by the Hawaii Department of Agriculture. At this time, APHIS requires a one year quarantine, and HDOA is requiring a separate one year period, rather than running concurrently. Introducing key varieties has taken longer than the expected 1 year quarantine period.In varieties that are through quarantine, loss of plant inventory due to tissue culture contaminationhave dramatically reduced the number of plants available for propagation and subsequent field testing. Industry stakeholders are assisting grant personell in seeking a solution to this issue. The sudden retirement of the University of Hawaii PI under Objective 1b (micropropagation) has delayed the expansion of materials. This Objective has now been transferred to the Hawaii Agriculture Research Center. In Hawai'i, testing of fungal isolates under Objective 3B is still in thein vitroandin plantastages due to the sheer number of fungal isolates to test. Purduestruggled with repeatability and maintenance of in vivo CLR infections in the greenhouse under Objective 4Bbut have recently optimized the protocol for the successful in vivo maintenance of CLR in growth chambers. This protocol is being used to 1) obtain sufficient infected tissue samples for RNA extraction for genome annotation and differential experiments to find genes associated with virulence and that are specific to different races of CLR. 2) increase and maintain different races of CLR for subsequent analyses. Finding variable markers to study CLR population genetics has been difficult in Puerto Rico. The sole coffee extension agent in the state of Hawaii is currently on extended leave, which has slowed development of materials for growers. What opportunities for training and professional development has the project provided?Objective 1:Due to the delay in hiring full time technicians for this project. Limited Appointment hirings of college students have aided in getting the objectives complete while providing these students with training for their future careers. Objective 3:Field, greenhouse and laboratory experience in general microbiology and plant pathology, agricultural data collection, molecular techniques, mobile app technology, and data analysis have been provided to full- and part-time biological technicians and students working on the project in Hawai'i. Objective 4:A new post doc, Dr. Terry Torres Cruz, began in March 2023 at Purdue University. In Puerto Rico, undergraduate and graduate students, recent graduates, and a post-bac student from Hispanic Serving Institutions have been hired and trained at USDA-ARS in Mayaguez and UPR Rio Piedras on plant pathological techniques. These activities included various methods of plant propagation, DNA extraction from CLR pustules and coffee plants, inoculations procedures of CLR on susceptible coffee varieties, and inoculation techniques on leaf disks under laboratory conditions. Objective 5: a doctoral student is receiving training in survey development and data analysis How have the results been disseminated to communities of interest?Regular and ongoing activities have been conducted to facilitate communication and outreach to all communities of interest. Some of these Extension-specificactivities are listed under the "Other Products" Section. Consortium Project Investigators meet quarterly at an All-Team online meeting to discuss research successes and barriers. Minutes are kept, and subsequently shared. This provides regular contact and followup for issues between objectives and disciplines. Through June of 2023, growers, stakeholders, scientists and state agencies met on a semi-monthlyconference call organized by the Hawaii Coffee Association. In these calls, consortium members provide updates, field questions, and receive input on progress to ensure the research is meeting stakeholder needs. These calls averaged at least 30 attendees. The Project Manager attends the monthly Hawaii Coffee Association and Kona Coffee Farmers Association meetings, and briefs stakeholders on SCRI grant activities. What do you plan to do during the next reporting period to accomplish the goals?Objective 1A: Field Trials of CLR Resistant Varieties WCR IMLVT lines will continue propagating for field trials, and a few plants from each line will be acclimated to the greenhouse to send leaves for genotyping to verify the variety. These plants will also be used to confirm CLR resistance using leaf disc assays and for nematode screening (Objective 1C).Establish agreementswith tissue culture labs in the continental U.S. to import potential new cultivars following HDOA importation requirements. Objective 1B: Propagation of CLR Resistant Varieties Due to the unexpected retirement of the PI for this sub-objective, this objective will be carried out further through the Hawaii Agriculture Research Center (HARC). Cultures that were started at UHH will be acclimatized and send out for field testing to be compared against Kona Typica plants. HARC has established cultivars of Obata into somatic embryogenesis media. We are working with breeders to get permission to propagate the IMLVT lines for the Hawaii coffee growers. We also will be looking at getting material in from commercial tissue culture laboratories in the continental U.S. to test other varieties. Objective 1C: Root-knot Nematode Resistance Trials and Enhancement of Resistance Screening Additional nematode bioassays will be conducted as more material from World Coffee Research and other sources becomes available.Nematode surveys will be conducted on coffee farms on the other Hawaiian Islands. Objective 2: Conduct third year of monitoring at all sites to capture differences in CLR biology, coffee plant phenology, management practices, and weather. Objective 3A: Field and laboratory research to identify effective and safe fungicides Conduct third year of field trials to capture variation across multiple seasons. Evaluation of rotation programs with the use of preventive and systemic fungicides for the full coffee season. Continue testing the efficacy of fungicidesin vitro,in planta, and in the field. Objective 3B: Continue testing the fungal isolates from Hawai'i for antagonistic behavior towards CLRin vitroandin planta. Begin evaluating mixes of the best biological control species/isolates from Puerto Rico against CLR. Objective 3C. Optimizing Field Health for CLR Management in Non-Resistant Varietals Continue visiting farms regularly to document all agronomic practices and CLR management.Support coffee farmers with specific recommendations for improving coffee plant nutrition based on two rounds of soil/leaf tissue sampling, previous fertilization records, elevation, planting density, tree age, and time of year. Conduct comparisons and analysis on the differences in agronomic practices and how this affects CLR. Objective 4A: We are collaborating with Dr. Rachel Koch at USDA ARS Ft. Dietrick to genotype and generate complete genomes for other races. This first comprehensive genome and additional genomes from other CLR strains using Nanopore will be analyzed for comparisons. Objective 4B: Initiate inoculations of CLR on plants to conduct gene expression studies. Objective 5:The plan is to use the survey data to conduct economic analysisto better understand the costs,benefits, and tradeoffs across different control strategies from the perspective of coffee growers and the larger coffeeindustry.

Impacts
What was accomplished under these goals? In 2022, the first year of the grant and second full year of Coffee Leaf Rust infection, Kona coffee growers reported a 50% reduction in yields due to severe defoliation from CLR. In 2023, leaf counts and subsequent yields rebounded close to average levels, significantly improving the economic situation for growers.Researchresults and extensive tech transfer under SCRI funding led to agronomic refinements in the field which facilitated this recovery.Through proper timing and use of fungicidal sprays, infection damage was reduced and crop loss was stabilized. As we learn more about the fungus' behavior in both Hawaii and Puerto Rico, and see the economic burden of maintaining susceptible trees, growers are increasingly interested in resistant varieties. Stakeholders have taken a proactive role in this project, assisting scientists with identifying varieties currently in the field and communicating their preferences on the taste profile.One producer located three trees with apparent resistance in his field and alerted ARS.The work under this grant has broad visibility in the grower community. Objective 1A: Propagation of the World Coffee Research (WCR) International Multi-Location Variety Trial (IMLVT) coffee lines is continuing. Inventory of the lines decreased due to general plant tissue culture contamination that occurred once the plants were propagated after quarantine. Select lines from the quarantined material are being acclimated in the greenhouse and/or placed in somatic embryogenesis media. In addition, in order to supplement the cultivars that have low inventory, we have imported coffee seeds and in vitro plantlets from Costa Rica under an APHIS Controlled Import Permit. Objective 1B: A Hawaii grower identified three trees that appeared resistant to CLR in his field. Laboratory tests confirmed resistance toHemileia vastatrixrace XXIV. Genotyping of these lines was completed by Dr. Dapeng Zhang, USDA ARS Beltsville. Somatic embryogenesis derived from leaf tissue was used to establish the plants in tissue culture. Objective 1C: Species-specific primers forM. konaensiswere developed to remove the sequencing step.A sterile culture was established to enable early screening of coffee cultivars for nematode resistance in tissue culture. Objective 2: Monthly surveys were conducted in 15 Hawaii farms and 10 farms in Puerto Rico.Data included CLR incidence and severity, CLR life stage, spore germination/viability, weather data, management activities, spore dispersal via wind and leaf counts. Average CLR incidence is high during the harvest and post-harvest (Aug-Feb, ~30%) and low during fruit development (Mar-Jul, 5-10%). Incidence and severity are positively correlated. No significant difference observed in incidence at varying elevations. Severe defoliation was observed from Nov 2022 through March 2023, followed by a new leaf flush. Farms in Kona are estimated to have lost 30-75% of yields in 2022, with a 18-30% recovery estimated for 2023. Spore capture is positively correlated with incidence. Spore germination is highest in shade vs. sun and on low vs. mid branches. Objective 3A: This concluded the second year of afield trial conducted in five farms testing efficacy of seven products registered for use and commonly available to growers in Hawaii. This included copper products, biologicals, the translaminar Priaxor, and a biostimulant.In addition, aYear-2 trial consisted of 5 treatments of three unregistered systemic fungicides containing Azoxystrobin, Picoxystrobin, Myclobutanil (with subsequent crop destruction) and 2 commercially available biological control products, Extract ofReynoutriasachalinensis, andPseudomonaschlororaphisstrain AFS009, plus an untreated control. Methodology for testing compatibility of CLR-targeted fungicides and Coffee Berry Borer pest-targeted BotaniGard was developed. Fourteen products were tested, resulting in 54 different combinations of product rates. Tank mixing experiments were conducted twice per product. Results showed thatBiological products provided protection for 4-5 weeks, while Copper products protected for 5-8 weeks and Priaxor (translaminar) protected leaves for 8-10 weeks.On the single organic farm examined, the botanical-based ProBlad fungicide did not outperform the foliar biostimulant CafeDak. Priaxor sprays provided better protection when conducted 8 wks apart vs. the label recommendation of 3 wks apart.The unregistered systemic fungicides all showed excellent control for extended periods. Objective 3B: Isolations of fungi from coffee leaves infected with CLRwere purified and identified using morphology and/or PCR and sequencing.Tree age and variety was also collected as well as monthly precipitation data.In total, 203 isolates were purified and identified comprising 43 genera. The number of isolates placed into long term storage for future study was 128. The isolateClonostachysshows promise, and more work is needed to verify the activity of this and remaining isolates. In Puerto Rico an experiment was completed on the effect of the fungal hyperparasiteLecanicilliumon CLR lesions. Significant effects were found, but strain efficacy differed by site. Objective 3C: Evaluations of agronomic practices and management of CLR were conducted on 11 farms.Monthly evaluations of CLR incidence, fungicide applications, weed control, pruning style, coffee production, and soil/leaf tissue samples. Coffee farms in which agronomic practices are conducted properly have shown lower CLR incidence in comparison with poorly managed farms. Data show that pruning styles significantly impact infection rates. During the full coffee season, coffee farms that only applied preventive fungicides (5-10 times) had high CLR infection (5-45%) in comparison with coffee farms that used a combination of the translaminar fungicide Priaxor (1- 2 times) with preventive fungicides (4-6 times) resulting in low CLR infection (0.5- 5%). Objective 4A. Hemileiavastatrixspores and CLR symptomatic leaves were collected from various locations on Hawaii Island and transferred to Purdue for genetic studies. The CLR genome was sequenced using three different platform technologies to obtain a comprehensive and well assembled genome. The genome for the Panama strain was assembled using HiFi reads and completed haplotype-phasing since urediniospores are dikaryotic, with 750 Mb for one nucleus and 724 Mb for the other. RNA was collected in the field for the Panama strain to aid in genome annotation. We have 2 Nanopore runs for the HI strain for comparison and are in the process of conducting RNA-sequencing on it. Population analyses of Hawai'i isolates using 12 SSRs has continued. Sampling for this analysis is nearly complete excepting some additional samples from Puerto Rico. Key historical CLR samples have also been added to this dataset and we are genotyping 4 CLR races obtained from USDA Ft. Dietrick. In Puerto Rico, DNA has been extracted from CLR collections from several sites and cultivars and microsatellite loci amplified. New candidate microsatellite loci have been tested.The microsatellite loci have shown little variation so far, and then new microsatellites have amplified well but have not been variable. Objective 4B: Puerto Rico obtained and planted coffee seeds of susceptible ('Caturra' and 'Catuai') and resistant local varieties ('Limani' and 'Fronton"). CLR inoculation on in vivo plants is a slow process. CLR infection was obtained onC.arabicacv. Caturra under laboratory conditions. Objective 5: Economics The coffee grower survey was launched in April using the membership list of various coffee organizations active in Hawaii. Response to the survey was very low for the first 3-4 months. Efforts were intensified to reach farmers in late summer, and we have now received more than 80 responses, with a goal of 100 by November 2023.

Publications

  • Type: Journal Articles Status: Published Year Published: 2023 Citation: Keith, L. M., Matsumoto, T. K., Sugiyama, L. S., Fukada, M., Nagai, C., Pereira, A. P., Silva, M. C., and V�rzea, V. 2023. First report of the physiological race XXIV of Hemileia vastatrix (coffee leaf rust) in Hawaii. Plant Dis. 107:2528. doi: 10.1094/PDIS-03-23-0460-PDN.
  • Type: Journal Articles Status: Published Year Published: 2023 Citation: Urbina H & Aime MC. 2023. The first USA continental record of coffee leaf rust (Hemileia vastatrix) on coffee (Coffea arabica) in southwest Florida, USA. Plant Dis. doi: 10.1094/PDIS-09-23-1869-PDN.
  • Type: Journal Articles Status: Published Year Published: 2023 Citation: Myers, R., Mello, C., Nagai, C., Sipes, B., and Matsumoto, T. 2023. Evaluation of Coffee arabica cultivars for resistance to Meloidogyne konaensis. Agriculture 13:1168. https://doi.org/10.3390/agriculture13061168


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

Outputs
Target Audience:The primary audience target for Objectives 1, 2, 3, and 5 are coffee growers and extension agents. Specific outreach and extension efforts are described further under the Accomplishments: Communities of Interest section. Most projects are in their early stages. Data collected for Objective 1 will benefit the coffee industry in Hawaii and Puerto Rico by identifying coffee varieties resistant to CLR and other diseases. Data also will benefit coffee farmers in Hawaii because CLR race 2 (detected in Puerto Rico since 1990) is not present in Hawaii, and Puerto Rico can provide information to Hawaii about resistance and how to manage this CLR race 2 in case of this race appears in Hawaii. A stronger coffee industry impacts economically depressed areas like the mountain region of Puerto Rico and the Ka'u region of Hawaii by creating jobs in farms and coffee processing organizations. The genomic activities under Objective 4 are targeted to researchers and plant breeders. Changes/Problems:Both Hawaii and Puerto Rico have a one-year APHIS plant quarantine (PQ) process for coffee materials. Hawaii State Department of Agriculture has a secondary quarantine process once the plants have left Federal custody, which can extend the quarantine to up to two years. Delay of APHIS-PPQ permits due to administrative challenges (change in permit application processes at Beltsville) have slowed the Objective 1 segment of the project. Some plant lines have been able to clear Hawaii PQ. Once the quarantined coffee plants were allowed to be propagated, we experienced a loss of plants to contamination. This was especially problematic since 20 of the 30 ILMVT lines had only a few plants (less than 30) that were sterile at the start of the quarantine. Hurricane Fiona, which struck Puerto Rico in September 2022, heavily damaged fields and roads in the mountainous coffee region. Lack of access delayed survey and data collection through December. A portion of the harvest was lost, and trees were defoliated by high winds. Beneficially, not as many trees were lost in comparison to 2017's Hurricane Maria, and the region should recover relatively quickly. The Project Investigator, Dr. Michael Shintaku, leading Objective 1b is retiring unexpectedly. This activity revolves around tissue-culturing the new varieties. This will necessitate moving the micropropagation activities from the University of Hawaii to the Hawaii Ag Research Center, a 501(c)3 entity. The budget and work timeline will be nominally the same. What opportunities for training and professional development has the project provided?Due to the delay in hiring full time technicians for this project, Limited Appointment hirings of University of Hawaii college students for Objective 1 have aided in getting the objectives complete while providing these students with training for their future careers. Undergraduate and recent graduates from Hispanic Serving Institutions have been hired and trained at USDA-ARS in Mayaguez on plant pathological techniques. These activities included various methods of plant propagation, DNA extraction from CLR pustules, inoculations procedures of CLR on susceptible coffee varieties, inoculation techniques on leaf disks under laboratory conditions. For Objective 5, a masters student in Agricultural Economics is receiving training in survey development. How have the results been disseminated to communities of interest?On a semi-monthly basis, growers, stakeholders, scientists and state agencies meet on a conference call organized by the Hawaii Coffee Association. In these calls, consortium members provide updates, field questions, and receive input on progress to ensure the research is meeting stakeholder needs. These calls average at least 30 attendees. The Puerto Rico Agricultural Experiment Station held talks with coffee growers at two events. A poster will be presented in December 2022 at a local agronomy meeting. Two manuscripts are in preparation. Multiple talks were presented at the 2022 Annual Hawaii Coffee Association Conference, CLR virtual talk story sessions, and University of Hawaii (UH) Extension webinars. Information was also distributed through extension publications, visual aids, and during ongoing farm visits. UH Extension provided 20 educational opportunities to 711 coffee producers and stakeholders to learn about CLR best management practices, coffee pruning, sprayer calibration, safe pesticide use, and coffee propagation via workshop, webinar, and field day events. Translators were provided for events such as Worker Protection Standards, sprayer calibration, and pesticide use and publications are available in languages including Spanish, Ilocano, and Tagalog. Our HawaiiCoffeeEd.com website is frequently visited by over 12,500 unique viewers. Information at this site can also be translated to dozens of languages. We also hosted educational booth displays, presented during industry events and hosted a Virtual Invasive Pest Mini Conference with 149 attendees. "Farm doctor" visits were conducted throughout the state including on Maui, Oahu, and the districts of Kau, Kona, Hamakua, and Waimea on Hawaii Island. During the statewide Invasive Pest Mini-Conference and Kauai CBB and CLR Workshop, staff presented an updated best management practices recommendation for CLR management. The recording has also been viewed by 50 individuals. Overall feedback of this mini-conference was positive with 98% of participants reporting an increase in knowledge and understanding of the topic area and 81% intending to share learned information with at least one person. During the in-person coffee pest workshop on Kauai, a total of 17 producers participatedinthis workshop. 85% of attendees responding to a post-event survey reported a significant increase in knowledge. All learned at least one new thing, and more than 30% reported learning five new things that they would apply to their operation. What do you plan to do during the next reporting period to accomplish the goals?Objective 1A: Field Trials of CLR Resistant Varieties The World Coffee Research IMLVT lines will continue propagating for field trials in Hawaii and Puerto Rico, and a few plants from each line will be acclimated to the greenhouse to send leaves for genotyping at Beltsville to verify the variety. These plants will also be used to confirm CLR resistance using leaf disc assays and for nematode screening (Objective 1C). Puerto Rico will receive 11 coffee seed differential plants from Costa Rica (the total number of CLR differential plants is 27, Costa Rica only has 11) to obtain seedlings and to start CLR evaluations. In addition, CLR from Puerto Rico will be sent to Portugal for determination of CLR race. Seeds will be obtained from controlled pollinations of the 15 IMLVT lines to obtain "pure lines" for CLR testing. Cupping trials with Extension and coffee growers will be conducted, dependent on available supplies. Objective 1B: Propagation of CLR Resistant Varieties Due to the retirement of the PI for this sub-objective, this objective will be carried out further through the Hawaii Agriculture Research Center (HARC). Established cultures will be sent from the University of Hawaii Hilo facility to HARC in two shipments to ensure plants are not lost to contamination during shipment. The first shipment will be verified to be clean and alive before second shipment is initiated. Objective 1C: Root-knot Nematode Resistance Trials and Enhancement of Resistance Screening Once IMLVT plants are in the greenhouse, screening for nematode resistance will commence. Complete the statewide survey on nematodes in Hawaii. Objective 2: CLR Surveys and Environmental Monitoring Conduct the second year of monitoring at all sites to capture differences in CLR biology, coffee plant phenology, management practices, and weather. Repeat spore viability assays and conduct potted plant experiments to assess additional environmental parameters. Utilize results to develop comprehensive IPM recommendations for Extension activities with growers in both Puerto Rico and Hawaii. Updated Puerto Rico recommendations are expected to go to print by February with field days shortly after. Hawaii IPM recommendations will be narrowed down through an "IPM Summit" with researchers, extension staff, and stakeholders in January with documents and field days shortly after. Objective 3A: Field and laboratory research to identify effective and safe fungicides Continue potted plant bioassays. Conduct second year of field trials to capture variation across multiple seasons. At the Kona Hills field research location, continue to collect monthly efficacy data to complete the first year field trial, analyze data for entire first-year efficacy trial, and repeat the field trial for a second year in order to reach more conclusive conclusions. Objective 3B: Biological Control of CLR Write the first manuscript on biological control of CLR in Puerto Rico; do additional experiments. From the data collected, evaluate the biocontrol potential of Hawaii isolates. Write the first manuscript on biological control of CLR in Hawaii. Objective 3C. Optimizing Field Health for CLR Management in Non-Resistant Varietals Maintain regular farm visitations to document all agronomic practices and CLR management. Support coffee farmers with specific recommendations for improving coffee plant nutrition based on two rounds of soil/leaf tissue sampling, previous fertilization records, elevation, planting density, tree age, and time of year. Conduct comparisons and analysis on the differences in agronomic practices and how this affects CLR. Objective 4A. Genetics of CLR Currently, the Purdue Aime Lab is cleaning the reads and conducting initial QC. With the arrival of a post-doc researcher at Purdue in the spring of 2023, the completed genome will be assembled and annotated and then run through genomic and population analyses.This comprehensive genome will form the scaffold for comparison for additional genomes from other CLR strains, to be generated on a single platform and analyzed. Developed protocols will be used to obtain sufficient infected tissue samples for RNA extraction. RNA will be used to annotate the genome and will form the basis of differential experiments for finding genes associated with virulence and that are specific to different races of CLR. Greenhouse protocols will be used to increase and maintain different races of CLR for subsequent analyses.Hawaii and Puerto Rico staff will continue to collect, store and transfer H. vastatrix spores to Purdue as needed. Objective 4B: Understanding Resistance to CLR Initiate inoculations of CLR on plants to conduct gene expression studies. Objective 5: Economics of CLR Launch the grower survey in January 2023 using the membership list of various coffee organizations active in Hawaii. Data collected from this survey will be used to conduct economic analysis to better understand the costs, benefits, and tradeoffs across different control strategies from the perspective of coffee growers and the larger coffee industry.

Impacts
What was accomplished under these goals? In 2022, the second full year of Coffee Leaf Rust infection, Kona coffee growers are estimated to have lost 30-75% of their crop. As early research results come in, there is a strong sense of urgency in the scientific and grower communities to address immediate tree health solutions for susceptible trees. First year results point to potential agronomic refinements in the field, as well as proper timing and use of fungicidal sprays, that may reduce infection damage and stabilize crop loss. As we learn more about the fungus' behavior in both Hawaii and Puerto Rico, and see the economic burden of maintaining susceptible trees, growers are increasingly interested in resistant varieties. Stakeholders have taken a proactive role in this project, assisting scientists with identifying varieties currently in the field and communicating their preferences on the taste profile. The work under this grant has broad visibility in the grower community. Objective 1A: Field Trials of CLR Resistant Varieties Hawaii ARS scientists are propagating the World Coffee Research (WCR) International Multi-Location Variety Trial (IMLVT) lines for field trials. To expedite the trials at Mayaguez, WCR granted access to 15 of the IMLVT Puerto Rico lines. All lines were verified by genetic analysis through Dr. Dapeng Zhang at USDA ARS Beltsville. In addition, permits were obtained from APHIS-PPQ and PR Department of Agriculture to introduce seed and in vitro plantlets from Costa Rica and Hawaii. The ARS Mayaguez facility installed growth chambers and bio safety hoods for CLR inoculations and growth of in vitro lines. Objective 1B: Propagation of CLR Resistant Varieties Both regions require a 1-year minimum quarantine period for inbound plant materials. To hasten availability, Hawaii growers identified CLR-resistant varieties already present in the field. Resistance and genotyping were confirmed by ARS scientists, and leaves were obtained for somatic embryogenesis propagation. Callus and direct somatic embryos have been observed in the cultures and are consistent with published literature. Many versions of media were tested as published recipes vary, with success dependent on the coffee variety. Objective 1C: Root-knot Nematode Resistance Trials Resistance screening is underway with faster screening methods under review. Objective 2: CLR Surveys Sampling was conducted every three weeks at 15 sites in Puerto Rico and every four weeks at 15 Hawaii farms. Active and passive spore samplers were installed in Hawaii to characterize pathogen movement. To assess the lifecycle, H. vastatrix spores were collected bi-monthly and compared. CLR infection levels fluctuate in Puerto Rico. Leaves on upper branches have less CLR than middle and lower branches. Highest levels of infection surprisingly appeared in summer but the plots may contain cultivars that differ in susceptibility. Research continues. For Hawaii, average CLR incidence was low early in the season (~4%, Jan-Jun) and increased to ~36% during the harvest (Jul-Dec). Incidence and severity are positively correlated. No significant difference in incidence due to elevation was observed, although farms at mid to high elevations had increased harvest incidence. Severe defoliation was observed from Nov-Mar, followed by a new leaf flush. Results from viability studies ranged from ~20-70% and suggests that spore germination rates vary depending on collection site, time of year, and management practices utilized. Germination was highest in shade and on lower branches and increased during the harvest season. Results from potted assays suggest that low spore density can result in symptom development and relative humidity plays only a minor role. Objective 3A: Field Management Research Fungicide protection experiments were conducted on potted seedlings in a growth chamber. Plants were inoculated with spores on day 0, 7, and 14 days post application (DPA) of the product. Control plants had water applied to the foliage. Trials on three farms tested the efficacy of four commercially available fungicides: Kocide, Badge X2, ProBlad Verde, and Serenade. Another field trial was established for fungicides not yet labeled in Hawaii: 3 systemics and 2 biological control products (Azoxystrobin, Picoxystrobin, Myclobutanil, Extract of Reynoutria sachalinensis, and Pseudomonas chlororaphis strain AFS009). Three replications of each treatment and control were included and organized in a randomized complete block design. For all trials, disease incidence and severity ratings were collected. For potted plant experiments, water-only positive controls sporulated between 14-35 days and had the highest disease incidence ratings. Treated plants had low disease incidence ratings 7 to 14 DPA of the treatment and provided protection against CLR infection when applied proactively. At the first field trial, all products provided protection for 5-6 weeks after the sprays, with variable efficacy levels.Field data suggests that all treated plots are experiencing less spread of CLR in comparison to the untreated control. As of September 2022, the systemics (severity under all 3: <1) are working most effectively. Untreated plots have more than double the amount of disease. No phytotoxicity has been observed. Objective 3B: Biological Control In Puerto Rico, various Lecanicillium-type fungi have been isolated from CLR lesions and inoculated in the field in three locations under two different seasons. Species varied in survival and growth of CLR lesions after application. The experiment will be repeated. In Hawaii, symptomatic leaf samples were collected from various locations, including an organic farm. More than 75 isolates from multiple fungal genera were recovered. The three most common isolates included the following genera: Simplicillium/Cephalosporium, Akanthomyces/Lecanicillium/Verticillium, and Cladosporium. Methods for mycoparasite efficacy were assessed and preliminary biocontrol evaluations of four isolates look promising. Objective 3C. Optimizing Field Health in Non-Resistant Varietals Monthly evaluations of agronomic practices and CLR management were conducted on 11 Hawaii farms. Fields show a high variability in terms of cultivars, elevation, pruning styles, topography, planting density, age of trees, weather conditions and agronomic practices. In cooperation with growers, records of all agronomic practices are documented as well as soil and leaf samples. CLR incidence ranged between 0.1% and 60%. In the early coffee season (April- July), CLR was on average < 3% with slow increases, while during the harvest season (Aug-Dec) CLR increased faster (3-9%). Across the entire season, block pruning systems showed the lowest CLR infection (0.8-2.2%), while traditional Kona style pruning averaged 12.4%. Coffee fields that only applied preventive fungicides had higher infection (12.4%) in comparison to fields in which both fungicides (preventive and systemic) were applied (1.4%). In all farms, recommendations for the improvement of agronomic practices were discussed with growers. Objective 4A. Genetics of CLR To obtain a comprehensive and well assembled genome, the Purdue Aime Lab sequenced the CLR genome using three different platform technologies: Illumina short reads; long-read sequencing at the UC Davis Genome center for PacBio; and long reads using Oxford Nanopore. The Lab have successfully defined protocols for the highly challenging in vivo maintenance of CLR in the greenhouse. Objective 4B: Understanding Resistance Puerto Rico has planted verified true-to-type seeds of susceptible and resistant local varieties to start inoculations of CLR and to conduct gene expression studies. Objective 5: Economics Economists attended the annual meeting of the Hawaii Coffee Association and met with consortium members, industry leaders, and growers to understand the challenges of CLR. Based on this, a questionnaire was developed and is currently being pre-tested.

Publications

  • Type: Journal Articles Status: Published Year Published: 2022 Citation: Aristiz�bal, L.F.; Johnson, M.A. Monitoring Coffee Leaf Rust (Hemileia vastatrix) on Commercial Coffee Farms in Hawaii: Early Insights from the First Year of Disease Incursion. Agronomy 2022, 12, 1134. https://doi.org/10.3390/ agronomy12051134
  • Type: Journal Articles Status: Published Year Published: 2022 Citation: Kawabata, A.M., S. Wages, and S.T. Nakamoto. (2022) Pruning methods for the management of coffee leaf rust and coffee berry borer in Hawaii. Honolulu (HI): University of Hawaii. 9 pp. (Plant Disease; PD-126).
  • Type: Other Status: Published Year Published: 2022 Citation: Baur M.E., Kawabata A.M., Nakamoto S.T., Shriner S., and Elliott S. 2022. National Pest Alert: Coffee leaf rust Hemileia vastatrix. North Central IPM Center.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: 1. Kawabata, A.M., M. Miyahira, S.R. Sand, and S.T. Nakamoto. 2022. Managing the Risks from Coffee Leaf Rust, a New and Devastating Disease Affecting Hawaii Coffee Producers presented at the 2022 Extension Risk Management Education National Conference, Omaha, Nebraska. 30 March 2022. (Poster presentation). https://agrisk.umn.edu/Conferences/Presentation/managing_the_risks_from_coffee_leaf_rust_a_ne
  • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: 2. Uyeda, J., A.M. Kawabata, M. Miyahira, S. Sand, and S.T. Nakamoto. 2022. Developing Sprayer Calibration Tools for Coffee and Orchard Crop Farmers that Reduce their Production and Legal Risks presented by Uyeda at the 2022 Extension Risk Management Education National Conference, Omaha, Nebraska. 30 March 2022. (Poster presentation). https://agrisk.umn.edu/Conferences/Presentation/developing_sprayer_calibration_tools_for_coff