Brief Summary of Minutes of Annual Meeting

Studies of microbial communities in CA, MN, NY, OH, and WA have indicated signficant variations in soil and rhizosphere communities associated with soilborne disease development and plant disease suppression. In CA and OH, such approaches have already led to the discovery of new biocontrol agents. Future research will focus on better characterizing those differences and how community structure is affected by various farm practices, e.g. incorproation of green manures. Several new collaborations have developed in the past year among various group members. Discovery research in MN, NE, OH, and WA has led to the discovery of new and useful biocontrol agents. Future work will aim towards a better understanding of the contributions of those microbial agents to plant disease suppression and the development of such strains, and/or their secretion products, into new biopesticides. Research by multiple groups on green manures indicated their general utility for enhancing natural biological control activities. Details of which amendments work best will be the focuse of future research by mutlipel groups. Practical field trials in CA, MI, NM, NY, and OH indicate the need to extensively field trial biopesticides. Data on Contans was generally positive, and its value for controlling Sclerotinia diseases was shown. While companies will pay for some contract research, information on proper application and efficacy in regional cropping systems is required to serve grower stakeholders most effectively. NY researchers have developed an approach to identify sources of Phytophthora contamination in nurseries. This information provides a scientific basis for the development of Best Management Practices that will be effective in eliminating sources of contamination. Multiple research groups in this project have found evidence for the multifactorial nature of plant health promotion, further expanding our understandin of biocontrol in the broadest sense. Such evidence, has led to new hypotheses about the nature of plant-microbe interactions and may lead to a shift in the paradigm currently guiding biological control research. Multiple members of this committee (NY, OH, WA) have been actively engaged in the development of extension publications, presentations, and workshops that promote the science-based utilization of cropping practices and inputs that reduce soilborne diseases. Such work provides stakeholders with the information and tools they need to increase crop productivity. Objective 1 To identify and characterize new biological agents, naturally suppressive soils, cultural practices, and organic amendments that provide control of diseases caused by soilborne plant pathogens. CA: Heterodera schachtii suppression. We determined that the population densities of the H. schachtii, two nematode generations after infestation, were dependent on the population densities of Dactylella oviparasitica and H. schachtii at the time of infestation. ITS analysis of our suppressive 9E field soil identified a new strain of D. oviparasitica. CA: Characterization of biological basis for peach replant disease. We identified two stramenopile phylotypes that negatively correlated with peach plant weights (Bent et al. 2009b). These organisms were Pythium ultimum and Sellaphora spp. MI: Isolation of beneficial bacteria for controlling soilborne diseases.From the suppressive soil, we have isolated 6 Bacillus spp. that have strong antagonistic characteristics against multiple soilborne pathogens, including Phytophthora spp., Sclerotinia sclerotiorum, and Streptomyces spp. including S. scabies.. These isolates have shown to reduce the severity of potato common scab in the greenhouse, which will be repeated next year. MI: Characterization of disease suppressive soil against potato common scab. We have identified a field that has suppressive soil inhibiting potato common scab in Michigan. This disease suppressiveness was verified to be biological by mixing the soil with autoclaved soil at different ratios or treating the soil with various temperatures, with Streptomyces scabies inoculated in the soil before the treatment. The soil profile was studied using terminal restriction fragment length polymorphs (T-RFLP), and conventional dilution plating using different selective media. Primary data have shown that Bacillus spp., and Streptomyces spp. are to be responsible to the disease suppression. MI: A new strain (DS3024) of Streptomyces sp. has been isolated. It was confirmed as a pathogen, but genetically different with any known pathogen Streptomyces species that cause potato common scab. It has txtAB genes that are associated with thaxtomin synthesis, which is a key factor of pathogenesis of the pathogen, but lacks of nec1 gene, which is common in most of the other pathogenic Streptomyces. It is possible that the pathogenic gene cluster has migrated into a new species that is naturally non-pathogenic. This has lead us to survey the Michigan potato production area for more population information: whether this strain is widely distributed and any new species exists in the soil. NM: Seed treatment with biofungicides for control of Phytophthora blight on chile pepper. A field study was conducted to evaluate the effect of seed treatment with two biofungicides (Micro108 and Kodiak), a chemical fungicide (Apron XL LS), a plant activator (Messenger), and a plant extract (Capsicum oleoresin at 5% aqueous solution) on development of Phytophthora blight (Phytophthora capsici). Seeds not treated (control) were included in the study. There was no significant difference among treatments with respect to weights of marketable pods and incidence of wilted plants. Seed treatment alone was insufficient in providing season-long protection to chile pepper against Phytophthora blight and other soilborne diseases. This result strengthens the conclusion of 2008 research. NM: Use of graminaceous green manures for control of Verticillium wilt on chile pepper. A greenhouse study was conducted to evaluate the effect of green manures from field corn, sorghum, sorghum sudangrass, pearl millet, wheat, and oat on Verticillium wilt (Verticillium dahliae) of chile pepper. Severity of Verticillium wilt was most reduced when infested soil was amended with sorghum sudangrass, wheat, or oat green manures. Graminaceous green manures have the potential to reduce Verticillium wilt on chile pepper. NY: Detection and identification of plant pathogens. A membrane-based multipathogen detection macroarray has been developed. The array can detect over 40 fungal and oomycete plant pathogens from plant tissue and soil. We are currently optimizing methods to detect pathogens from irrigation water. The array is also being used to identify pathogens in the rhizosphere following various mixed-species green manure treatments. NY: Effects of selected grain crops on survival and infectivity of Rhizoctonia in field microplots: We reported last year that a number of isolates of Rhizoctonia and Rhizoctonia-like fungi obtained from naturally infected vegetable crops were capable of infecting corn. A greenhouse test also showed that the most aggressive isolates to corn were also highly aggressive on beans. A field microplot test was initiated in the summer of 2009 to determine the influence of other grain crops (rye grain, sweet corn, wheat and sudangrass) on the survival and infectivity of three isolates of Rhizoctonia (R-39, AG 2-2; R-43, AG 4; and R-62, Binucleate). The crops were planted in microplots established in an experimental field at the vegetable Research Farm. Seeds of snap bean cv.Caprice were planted in the microplots and maintained for 30 days. Results obtained showed that all the isolates in all the treatments were capable of infecting bean. NY: Effect of selected cover crops on root rot severity and yield of dry beans: Seven cover crop treatments (rapeseed, canola, grain pearl millet, rye/vetch, buckwheat, sudangrass and a fallow check) were established in 2007 in a section of our experimental bean root rot field at the Vegetable Research Farm of the NYSAES. After two cycles of growing these crops, plots were planted to dry bean cv.CLRK in June 2009 and root rot severity and yield were assessed. At planting time, soil samples were collected from each plot for conducting the bean bioassay in the greenhouse. In the greenhouse bioassay, root rot severity was reduced significantly only on roots of bean growing in soil planted to a cover crop of sudangrass (Table 2). However, no significant reduction was observed on dry bean grown in the field plots nor there was difference in bean yield (Table 2). The latter suggest that two cycles of cover crops incorporated as green manures are not enough to significantly reduce root rot severity caused by a complex of root pathogens that prevail at this site (Rhizoctonia, Fusarium, Pythium and Thielaviopsis). NY: Additional beneficial effects of Trichoderma agents. We have demonstrated that biological control is only a subset of the capabilities of Trichoderma strains for use in plant agriculture. They also control physiological and abiotic stresses such as drought, salt and other factors. They also increase nitrogen use efficiency in crops. This is a very large potential benefit since we think we can reduce nitrogen fertilizer by 30% and still improve plant yields. All of these advantages, including plant disease control, are brought about by the ability of the fungi to colonize plant roots, establish chemical communication with the plant and induce systemic reprogramming of plant gene expression. OH: Marker-assisted recovery and characterization of novel biocontrol bacteria. We completed our work on identifying some the components of soilborne disease suppression mediated by a mixed hay cropping in a transitional organic cropping system. Using terminal restriction fragments (TRF) statistically associated with the mixed hay-induced suppressiveness as probes, we were able to recover eight isolates representing the M139 and M141 marked populations. Sequence analyses showed the M139-marked strains to be a novel species most closely related to Mitsuaria chitosanitabida. The M141-marked strains were found to represent a species of Burkholderia not previously cultured. Both sets of strains were able to inhibit a broad spectrum of soilborne pathogens in vitro, and they were also able to suppress lesion development in planta. WA: A diversity of novel phenazine producers is predominant in dryland wheat soils. The biogeography of phenazine producing Pseudomonas spp. was investigated in the large dryland wheat growing region of the inland Pacific Northwest. High populations of phenazine producing strains were found in the low precipitation areas, but not the higher precipitation areas. Irrigation in dryland areas completely shifted populations to phloroglucinol producers. High levels of phenazine were detected in the rhizospheres of plants sampled in the field. Strains were characterized based on phzF and 16S genes. The main group of Pseudomonas clustered with P. orientalis, distinct from 2-79, which clustered with another group - P. synxantha, P. gessardii, and P. libanensis. Strains were genotyped with box PCR and phenotyped with Biolog. 31 genotypes were identified, which clustered into four groups. WA: Limestone amendments suppress Fusarium wilt in spinach seed crops. A field trial was set up to evaluate whether annual applications of limestone amendment at 1 or 2 tons/acre can suppress Fusarium wilt adequately and in an economically-viable manner, to enable a 4-5 year rotation between spinach seed crops. Extensive data was collected for each treatment combination on soil and plant nutrient status, spinach biomass, disease development, seed and soil infection levels by Fusarium and Verticillium (another wilt pathogen of spinach), etc. Each season, soil samples will be assayed for changes in chemical and microbial populations over the duration of the trial. Soil microbial analyses will be completed using 454-pyrosequencing. Objective 2 To understand how microbial populations and their gene expression are regulated by the biological (plants and microbes) and physical environment and how they influence disease. HI: Isolation of new bacterial biocontrol agents: A number of isolates of the bacterial species Stenotrophomonas maltophilia have been shown to act as biocontrol agents, particularly against fungal pathogens. Based on an extensive survey of Xanthomonas/Stenotrophomonas field isolates, we have selected a S. maltophilia accession isolated from cabbage seed for further study and comparative genomics efforts. This strain is being sequenced with the goal of assembling the entire genome and studying it with respect to genetic markers and phenotypes. MN: Pathogen suppressive activity was characterized for rhizosphere microbial communities associated with diverse native plant species in communities with varying plant diversity. Plant communities had been established as part of a long-term ecological research project in 1994. For individual plant species, pathogen suppression varied significantly when the plants were grown in low diversity (monoculture or 4-species) vs. high diversity (16- or 32-species) communities. Pathogen suppressive activity was also characterized for diverse plant communities across the landscape, including prairie, savannah, forest, and wetland sites. Finally, induction of pathogen suppressive activity by plant and microbial compounds was characterized for a diverse collection of microbial antagonists; induction tends to be highly specific, and was more likely among isolates from the same community. NE: Development of methods to more efficiently study Lysobacter sp. Strains of Lysobacter enzymogenes, a bacterial species with biocontrol activity, have been detected via 16S rDNA sequences in soil in different parts of the world. In this study, DNA-based detection and enrichment culturing methods were developed for Lysobacter spp. and L. enzymogenes, respectively. In the DNA-based method, a region of 16S rDNA conserved among Lysobacter spp. When L4-F and universal bacterial primer 1525R were used to amplify DNA from various bacterial species, a 1,100-bp product was found in Lysobacter spp. exclusively. The enrichment culturing method involved culturing soils for 3 days in a chitin-containing broth amended with antibiotics. A strain of L. enzymogenes added to soils was detected at populations as low as 102 and 104 CFU/g soil by PCR amplication with L4-F and the culturing method, respectively. When both methods were applied twice to assay 56 soil samples collected from various locations in Nebraska, Lysobacter was detected by PCR in 39, over 70%, of the samples, out of which 5 yielded strains of Lysobacter spp. by enrichment culture. Among 16 isolated strains, all were identified to be L. enzyomogenes, with the exception of a strain of L. antibioticus. OH: Characterization of a novel beneficial activity of DAPG-praoduicng bacteria. Application of the DAPG-producing strain Wood1R to seed prior to planting was found to ameliorate abiotic stress disorder when the corn grown in soils with a pH < 5.0. Under such conditions, there was no apparent increase in damping-off or visible root rots, indicating that soilborne pathogen suppression was not the primary mechanism leading to the enhanced plant growth. This work is the first to report the ability of a biocontrol strain to reduce an abiotic stress in crop plants, and suggest that additional investigations into the multivariate responses of plant hosts to biocontrol bacteria should be investigated. OH: Towards the discovery of a new genes involved in biological control The genome sequences of four bacterial biocontrol strains were obtained. Preliminary analyses reveal novel polyketide and lipopeptide synthases in the two DAPG-producing strains analyzed. In addition, putative insecticidal protein genes were found in Wayn1, indicating that the strain may also have potential for development as the source of new active ingredients for insect as well as disease control. OR: Ecology of Phytophthora ramorum: Chlamydospores of P. ramorum typically germinate at low frequency in the lab. We demonstrated that the frequency of chlamydospore germination is increased by passage through the gut of the Pacific banana slug. P. ramorum sporangia and hyphae that pass through banana slugs also remain viable. Under laboratory conditions, banana slugs can transmit ingested Phytophthora ramorum to leaves and tanoak boles. We do not know if slugs transmit P. ramorum in natural conditions. WA: Microbial communities in Rhizoctonia patches are different from surrounding soil: 454 pyrosequencing was used to compare bacterial communities inside and outside of Rhizoctonia bare patches, and in disappeared patches. Samples were taken from a continuous wheat rotation, with 4 blocks, from both bulk and rhizosphere soil. The most dominant group observed in ths study was the Actinobacteria, which accounted for about 45% of the sequences. In the rhizosphere, Pseudomonas, Enterobacteria, Sphingomonas, and Burkholdariales were most common. A number of taxa were in significantly higher rhizosphere frequencies inside of the patches, compared to outside of the patches. These included Flavobacterium, Microbacterium, Phyllobacterium, and Enterobacteriaceae. Fewer showed a tendency to high levels in the disappeared patch. These included Burkholdaria, Dyella, Friedmaniella, and Sphingomonas. Other common taxa were in Myxococcales, Verrucomicrobiales, Caulobacteriales, and TM7_genera_incertae_sedis. Surprisingly few Firmicutes were detected. Results will be confirmed with real-time PCR primers designed to these specific groups. WA: Mode of action of phloroglucinol is multi-faceted: Phloroglucinol is an antifungal compound produced by Pseudomonas spp., and is involved in the suppression of a number of pathogens on wheat roots, including Gaemannomyces graminis var tritici. However, little is known about the mode of action of this compound. This question was investigated, using a Saccharomyces yeast mutant library. This library has almost 5000 characterized ORFs annotated to function. We did not find any mutants with increased resistance, but a number of mutants were more sensitive. These could be classified into 3 main groups- ergosterol biosynthesis (membrane function), electron transport/oxidative stress, and V-ATPase synthesis and assembly. This suggests that phloroglucinol affects multiple basic cellular processes, and would be difficult to select for resistance in nature. Objective 3. To implement sustainable management strategies for soilborne pathogens that are biologically based and compatible with soil health management practices. CA: Evaluations of the biocontrol product Contans (Coniothyrium minitans) against lettuce drop caused by Sclerotinia minor in the Salinas Valley. Field experiments were continued to evaluate both the rates of Contans as well as the efficacy of timing and frequency of application to lettuce. In addition, laboratory experiments to evaluate the hypothesis that the most susceptible phase of S. minor is mycelial for attack by C. minitans were expanded to a range of isolates. The two rates of Contans were applied one week prior to, one wk after thinning, and finally before disking under the crop residue. Endura was applied immediately after thinning and at two weeks post-thinning. A single application of Contans at harvest had disease levels intermediate between the other Contans treatments and unsprayed control. While the lower levels of lettuce drop in Contans treatments were correlated with significantly lower levels of sclerotia, the lower levels of lettuce drop despite the presence of higher inoculum in the Endura treatment was attributable to the prevention of infection by S. minor. Tank mixes of Contans and Endura are incompatible. The ideal approach to lowering lettuce drop infections appears to be to employ Contans to lower the number of sclerotia in soil and Endura to prevent infection within a cropping season. CA: Assess the efficacy of various biopesticidal amendments to the recirculating nutrient solution for the suppression of root rot caused by zoosporic pathogens (Phytophthora capsici) and elucidation of the mechanism of suppression. With the exception of sodium salicylate, none of the other chemical amendments to the nutrient solution (i.e., Neem, xylene, zinc oxide, Perasan® or Actigard®) provided either consistent or significant control of the disease. All chemical amendments, except Perasan®, enhanced the total and the fluorescent bacterial population in the nutrient solution. However, the selective enhancement of the fluorescent pseudomonad population (which consisted of several biotypes) varied dramatically (0 to 87%) following each consecutive addition of sodium salicylate to the nutrient solution. The fluorescent pseudomonad population may or may not have contributed to the observed sodium salicylate-mediated suppression of the disease. MI: Efficacy trials of biopesticides for control of soybean white mold. This is the third year of trials using biological control to reduce the risk of soybean white mold. Contans (Coniothyrium minitans) had consistently showed the best efficacy, reducing Sclerotinia sclerotiorum sclerotia in soil by 95%. Actinovate (Streptomyces lydicus) had very similar efficacy as Contans on controlling the sclerotia. Potassium silicate had a significant reduction of S. sclerotiorum sclerotia and disease incidence, but less effective as the two mentioned biological control agents. These results were similar in two locations (Plant Pathology Farm and Clarksville). NY: Effect of tillage systems, cover crops and crop rotation on root rot severity and yield of peas: The long-term soil health site (about 14 acres) was established in 2003 to assess the effect of tillage systems (no-till, strip-till, and conventional tillage), three cover crops (rye grain, hairy vetch, and no cover/fallow) and 2 rotations (one heavy with vegetables, whereas as the other include grain and forage crops and less vegetables). All the treatments were replicated 4 times (4 blocks, 72 plots) and root health, soil health, and yield data have been collected annually. Root rot severity ratings of roots of peas grown in soil of all the plots were rather low and didnt exhibit the effect of the various practices In addition, growth parameter and yield of peas did not vary among the various practices, which could have been the result of the excessive rainfall that prevailed during moths June and July. The work on this site will be continued for the foreseeable future. NY: Utilizing green manures to reduce the severity of Phytophthora capsici on tomato. In 2009, the Smart lab started a project (in collaboration with the McSpadden-Gardener lab) to identify green-manure cover crops that are effective in reducing the incidence of Phytophthora blight on tomato. We have planted four mixed-species cover crops along with two controls (rye alone and bare ground). The crops will be tilled into the soil in spring 2010 prior to planting tomatoes on raised beds. There are two blocks (with 4 replicates of each treatment per block) so that treatments can be tested against two levels of pathogen inoculum (high and low levels of Phytophthora capsici). Plots will be rated for level of disease and yield. OR: Systems approach for producing nursery stock free of plant pathogenic Phytophthora species: We completed a 3-year project to develop a new approach for producing nursery stock free of Phytophthora that could also be applied to other pests and pathogens. Four Critical Control Points (CCPs) were identified: infested soil/gravel substrates, irrigation water, re-used pots, and potting media that has been contaminated by field equipment. The most common source of contamination in all tested nurseries was infested soil/gravel substrate in greenhouses and can yards. Healthy container plants set on these substrates readily became contaminated. We consistently recovered Phytophthora from recycled irrigation water and from containers designated for reuse. Our recommendations highlight the need to improve sanitation practices to target these sources of contamination in nurseries. OR: Characterization of pathogen diversity in nurseries. A total of 674 Phytophthora isolates was recovered from Oregon nurseries including 15 Phytophthora species and 7 other Phytophthora taxa. The most frequently isolated species from symptomatic plants were P. citricola, P. cinnamomi, and P. syringae. From gravel substrates, soil, and used containers, the predominant species were P. citricola, P. cinnamomi, and P. cryptogea. From irrigation ponds, most isolates were P. gonapodyides or other Phytophthora taxa belonging to ITS Clade 6. P. parsiana and P. inundata, two pathogens not previously reported from Pacific Northwest nurseries, were also detected. P. cinnamomi, the species most frequently isolated from plants, was never recovered from water. Phytophthora species recovered from specific CCPs differed among nurseries, likely a reflection of different production practices and composition of plant cultivars. WA: Brassica seed meals can suppress diseases in apple replant. Brassica seed meals can control control populations of Macrophomina phaseolina in the soil. These include Brassica juncea- (oriental mustard), Brassica napus (canola) and Sinapis alba (white mustard). However, this only occurred in natural soil. However, in pasteurized soil, these treatments did not suppress, indicating the requirement for resident soil microbes. Seedmeal also increased apple yield in apple replant problem trials, but only when used with Ridomil, since some seedmeals can stimulate Pythium. In an organic orchard, at the end of the third growing season, seedmeals gave similar growth to Telone C17. WA: Development of a soil bioassay for Fusarium wilt of spinach. Spinach seed growers can lose entire crops to Fusarium wilt despite rotations of 8-16 years between spinach seed crops. Research was initiated to develop a quantitative soil bioassay as a risk assessment tool for growers to select appropriate fields for seed crops to minimize losses to Fusarium wilt. The bioassay entails planting in a sample of soil from the field of interest a set of standard spinach inbred lines with different levels of susceptibility to Fusarium wilt, and monitoring the plants for Fusarium wilt using a quantitative rating scale. Based on promising preliminary results for a greenhouse soil bioassay in 2009, spinach seed growers and seed industry personnel have submitted soil samples from ~50 fields in northwestern Washington to test using the bioassay in the winter of 2009-10. The bioassay will be demonstrated to stakeholders in early March so they can potentially utilize the results in selection of fields for 2010 spinach seed crops. Those fields assayed and planted to spinach in 2010 will be monitored for Fusarium wilt. WA: Stunting caused by Rhizoctonia spp. in onion and pea crops following cereal cover crops in the semi-arid Columbia Basin. Empirical evidence suggests increasing problems in the semi-arid Columbia Basin of the Pacific Northwest with seedling blight in onion bulb crops and pea crops planted after incorporating cover crops such as wheat and barley. The problem occurs when onion or pea crops are planted in early spring soon after incorporation of a winter cover crop in strips, with the remaining cover crop left standing to protect emerging onion or pea seedlings from wind and sandblasting. Stunted patches range from <1 to >10 m in diameter. The problem does not occur in the absence of significant decomposing residues. WA: Pythium spp. associated with damping-off of organic vegetable crops in the semi-arid Columbia Basin of the Pacific Northwest.Growers and production managers of organic vegetable crops in central and western WA (representing production operations ranging from <5 acres to >6,000 certified organic acres) were interviewed to identify primary concerns regarding damping-off of organic vegetable crops. In summary, large-scale organic farms focused on processing vegetable crops experience the most significant losses to damping-off, primarily during early-season plantings. Many have to replant entire fields or seed at higher populations because of damping-off. None of the growers surveyed uses organic seed treatments because of inconsistent or no efficacy under field conditions. However, some indicated they would pay $100-$150)/acre for seed treatments if the treatments consistently controlled damping-off in cold, wet soils. Objective 4. Provide outreach, education, extension and technology transfer to our clients and stakeholders- growers, biocontrol industry, graduate and undergraduate students, K-12 students and other scientists NY: 2009 Cornell soil health train-the-trainer workshop in collaboration with the Cornell Soil Health program Work Team: This intensive soil health workshop included a mix of classroom training on the emerging concept of soil health, hands-on laboratory experience in assessment protocols, field assessment of soil health measurements, generating the soil health report and its interpretation, and a field trip to local farms to observe and discuss adopted soil health practices. Another workshop has been scheduled for August 15  19, 2010. Additional information on the workshops and the soil health program at Cornell can be found at our website (http://soilhealth.cals.cornell.edu). NY: Disease management strategies for Phytophthora capsici In 2009, Smart gave 13 talks to growers, extension educators and industry representatives on strategies to control Phytophthora blight. Additionally, a new web-site is being developed that will provide fact sheets that can be downloaded. This will provide specific information tailored to home gardeners, organic growers or conventional growers. NY: Outreach to K-12 students. We have continued our outreach program to third-grade students in the Geneva City School District (Geneva, NY). Part of this outreach includes a summer science camp, where students study different aspects of food production utilizing a garden that they plant at their school. One week of the 5-week program focuses on the importance of healthy soil to producing healthy vegetables. NY: Review of different models of biopesticide development and delivery. We are working on several models of production of biocontrol agents, ranging from classical registered pesticide, large national and international to small biovillage level systems. This topic is the subject of a feature article for Plant Disease that has been accepted with revision OH: Outreach and assistance to the biopestide industry. A presentation describing the utility of microbial community profiling for the discovery of novel biocontrol agents was presented at the Spring 2009 meeting of the BioPesticide Industry Alliance in Washington, DC. Additionally, contract research related to the reformulation and testing of a novel biopesticide was conducted in Ohio. Critical review and assistance was provided for the redevelopment of the Biopesticide Industry Alliance web site. OR: Development of science-based Best Management Practices for nursery growers. Our use of systems approach has been applied to determine sources of contamination in plant production systems. It will provide a basis for considering alternatives to the current system of end-point nursery stock inspections, and will guide in the development of targeted mitigation strategies. A novel and highly successful component of the project was the interaction between scientists and growers that resulted in development of science-based and practical Best Management Practices. OR: Online course on Phytophthora for nursery growers. In 2008, we launched a free online course to teach nursery growers about Phytophthora biology, symptoms, and disease management. Both English and Spanish language versions are available at http://ecampus.oregonstate.edu/workforce/phytophthora/ . In 2009, we had excellent feedback from the nursery industry as well as from college horticulture programs that use this in their curriculum. Several improvements to the course were made in 2009. WA: Extension outreach to vegetable growers: L. du Toit has a 40% extension appointment, and devotes much of her time to extension presentations and training growers, state department of agriculture personnel, consultants, and seed industry personnel in vegetable disease diagnosis and management. Lindsey presents her vegetable seed pathology research to stakeholders in the Pacific Northwest (ID, OR, WA), as well as to end users of vegetable seed produced in this region (e.g., AZ, CA, MI, Ontario) and international seed industry. She works with undergraduates and annually participaesat Mount Vernon High School to promote awareness of agricultural/science careers among youth in the Skagit Valley. L. du Toit and P. Okubara accommodated a WSU undergraduate intern, K. Reed, in each of their programs in 2009 to work on various molecular lab, greenhouse, and field aspects of spinach Fusarium wilt. WA: ARS scientists led a science outreach and engagement program to The Confederated Tribes of the Colville Reservation and the region surrounding Omak, WA. This is one of the poorest and underserved regions in Washington State. The program entitled "Pumping-Up the Math and Science Pipeline: Grade School to College" has four components: i) development of energy independence on the Colville Reservation through the production of biofuels; ii) hands-on science education in reservation schools by ARS research scientists, WSU faculty and Bellevue Community College instructors; iii) development of and participation in on-reservation summer science camps; and iv) employing Native American, Latino and rural high school summer science interns in ARS laboratories. Native American reservations and rural communities throughout the Pacific Northwest are requesting expansion of this ARS program to their communities.