Population structure and phenotypic variation of Sclerotinia sclerotiorum from dry bean (Phaseolus vulgaris) in the United States.

The ascomycete pathogen Sclerotinia sclerotiorum is a necrotrophic pathogen on over 400 known host plants, and is the causal agent of white mold on dry bean. Currently, there are no known cultivars of dry bean with complete resistance to white mold. For more than 20 years, bean breeders have been using white mold screening nurseries (wmn) with natural populations of S. sclerotiorum to screen new cultivars for resistance. It is thus important to know if the genetic diversity in populations of S. sclerotiorum within these nurseries (a) reflect the genetic diversity of the populations in the surrounding region and (b) are stable over time. Furthermore, previous studies have investigated the correlation between mycelial compatibility groups (MCG) and multilocus haplotypes (MLH), but none have formally tested these patterns. We genotyped 366 isolates of S. sclerotiorum from producer fields and wmn surveyed over 10 years in 2003-2012 representing 11 states in the United States of America, Australia, France, and Mexico at 11 microsatellite loci resulting in 165 MLHs. Populations were loosely structured over space and time based on analysis of molecular variance and discriminant analysis of principal components, but not by cultivar, aggressiveness, or field source. Of all the regions tested, only Mexico (n = 18) shared no MLHs with any other region. Using a bipartite network-based approach, we found no evidence that the MCGs accurately represent MLHs. Our study suggests that breeders should continue to test dry bean lines in several wmn across the United States to account for both the phenotypic and genotypic variation that exists across regions.

Genetic and aggressiveness variation among Sclerotinia sclerotiorum dry bean isolates from Brazil fields / Variação genética e agressividade entre isolados de Sclerotinia sclerotiorum provindos de campos de feijão do Brasil

Sclerotinia sclerotiorum, infection of bean fields, has increased in Brazil. Fungicides application is the control strategy used due to lack of cultivars with complete disease resistance. To guide the use of isolates in resistance screening 25 S. sclerotiorum isolates from Brazilian dry bean fields were characterized using microsatellite markers, mycelial compatibility groups (MCGs) and aggressiveness. Microsatellite primer pairs were used to identify polymorphisms among the S. sclerotiorum isolates and MCGs were determined from interaction of all isolates grown sideby-side. Aggressiveness was derived from a straw test where fungal mycelium was placed over a cut bean stem and rated for disease progress. Data from microsatellite profiles grouped the 25 isolates into four clusters and seven MCGs were identified. No association among host cultivar and cluster or MCG of isolates was observed. For MCGs, 57% contained isolates sampled frequently over multiple locations and 43% contained isolates unique to locations. There were significant differences among isolates in aggressiveness within and between MCGs. The most aggressive isolates in resistance screening will be helpful in the identification of higher levels of resistance in bean germplasm/lines.

A infecção de Sclerotinia sclerotiorum em campos de feijoeiro tem aumentado no Brasil. A aplicação de fungicidas é a estratégia de controle utilizada devido à falta de cultivares com resistência completa á doença. Para orientar o uso de isolados visando resistência, 25 isolados de S. sclerotiorum coletados em campos de feijoeiro no Brasil foram caracterizados utilizando marcadores microssatélites, grupos de compatibilidade micelial (MCGs) e agressividade. Pares de primers de microssatélites foram utilizados para identificar polimorfismo entre os isolados de S. sclerotiorum e MCGs foram determinados a partir de interação dos isolados crescendo lado-a-lado. O teste de agressividade foi derivado a partir do straw test onde o micélio do fungo foi depositado sobre a haste cortada de feijoeiro e avaliado o progresso da doença. Os dados de microssatélites dos 25 isolados de S. Sclerotiorum foram agrupados em quatro grupos e identificados sete MCGs. Não foi observada associação entre a cultivar hospedeira e o cluster ou MCG dos isolados. Para MCGs, 57% continham isolados amostrados em vários locais e 43 % continham isolados de apenas um local. Houve diferença significativa entre os isolados na agressividade dentro e entre os MCGs. O isolado mais agressivo no screening de resistência será útil na identificação de níveis mais elevados de resistência em germoplasma/linhagens de feijoeiro.

Uromyces appendiculatus in Honduras: Pathogen Diversity and Host Resistance Screening.

Bean rust, caused by the fungus Uromyces appendiculatus, is a major constraint for common bean production worldwide. Virulence of U. appendiculatus collected from wild and cultivated Phaseolus spp. was examined in 28 locations across Honduras. Host accessions representing wild and domesticated Phaseolus spp. collected at the same sampling locations were evaluated for resistance against U. appendiculatus. In total, 91 pathotypes were identified from 385 U. appendiculatus isolates according to their virulence on each of the 12 host differentials. No significant difference in pathogen total virulence, measured as the mean disease score, was found between locations. However, significant differences were found in pathotype virulence among isolates collected from different Phaseolus spp. within a location. Moreover, when locations were compared on the basis of pathotype occurrence and frequency, differences among locations were evident. No two locations had the same pathotype composition. The most common pathotype was virulent on 9 of the 12 differential lines. A high number of resistant accessions were identified in Phaseolus coccineus and P. lunatus. Although most wild P. vulgaris accessions were highly susceptible, rust resistance was observed in P. vulgaris landraces collected from farmer's fields. Thirty-two (52%) of the accessions screened showed intermediate to high levels of resistance and, of those, 16% were P. coccineus accessions. Our findings support the hypothesis that interaction of U. appendiculatus in host populations composed of diverse Phaseolus spp. and genotypes has favored highly diverse and virulent pathotypes, creating a center for virulence diversity of the pathogen in Honduras. The high percentage of intermediate and highly resistant accessions identified in the present study supports the strategy of collecting plants from the center of diversity of a pathogen or in locations with high incidence of disease and pathogen diversity to maximize the probability of identifying new sources of resistance.

Variation in Sclerotinia sclerotiorum Bean Isolates from Multisite Resistance Screening Locations.

There is no complete resistance to Sclerotinia sclerotiorum, cause of white mold in dry bean (Phaseolus vulgaris). Variable resistance expression is one problem in screening for improved white mold resistance. With no previous information in the literature, pathogen variation in multisite screening nurseries was evaluated as one cause of diverse resistance expression. In all, 10 isolates of S. sclerotiorum used in greenhouse screening and 146 isolates collected from nine white mold field screening nurseries in major bean production areas in the United States were compared using mycelial compatibility groupings (MCGs) and an aggressiveness test. These 10 greenhouse screening isolates formed six MCGs. Among 156 field and greenhouse isolates, 64 MCGs were identified and 36 of those were each composed of a single unique isolate. Significant differences in isolate aggressiveness were found between some isolates in different MCGs but the isolates within an MCG did not differ in aggressiveness. High isolate variation found within and between field locations could influence the disease phenotype of putative white mold resistant germplasm. We next compared genotype and phenotype of isolates from screening nurseries and those from producer fields. Variability found in and among screening locations did reflect variability found in the four producer fields sampled. White mold resistance screening can be improved by knowledge of isolate genotypic and phenotypic characteristics.

Evaluation of Resistance Screening Methods for Sclerotinia Stem Rot of Soybean and Dry Bean.

Three methods to identify levels of resistance to Sclerotinia sclerotiorum in soybean (Glycine max) and dry bean (Phaseolus vulgaris) were compared using multiple data analyses. The three methods were mycelial plug inoculations of cotyledons, cut stems, and detached leaves. Six S. sclerotiorum isolates of known relative aggressiveness were inoculated on each of three soybean and dry bean cultivars with varied response to S. sclerotiorum. For soybean, all three inoculation methods accurately identified isolate aggressiveness irrespective of cultivar, but identification of susceptible and partially resistant soybean cultivars was influenced by isolate. For dry bean, the cotyledon and cut stem methods accurately identified isolate aggressiveness, but identification of susceptible and partially resistant dry bean cultivars was influenced by isolate and inoculation method. The cut stem method had the smallest coefficient of variation and was more precise for detecting interactions. When considering root mean square residual error combined over species and experiments, coefficient of variation based on residual error, significance of isolate-by-cultivar interaction from ANOVA, rank correlation between pairs of methods, and sensitivity ratio for the three resistance screening methods under controlled environmental conditions, the cut stem method was statistically better than the cotyledon and detached leaf methods for evaluating resistance in soybean and dry bean cultivars.

Isolates of Uromyces appendiculatus with Specific Virulence to Landraces of Phaseolus vulgaris of Andean Origin.

Five isolates of the bean rust fungus Uromyces appendiculatus were shown to be specifically virulent on bean genotypes of Andean origin. This specificity was demonstrated by the virulence of five pairs of isolates on a differential set of 30 Phaseolus vulgaris landraces. Each isolate pair was from a different country in the Americas and consisted of one Andean-specific isolate and one nonspecific isolate. Of the differential P. vulgaris landraces, 15 were of Middle American origin and 15 were of Andean origin. The Andean-specific rust isolates were highly virulent on Andean landraces but not on landraces of Middle American origin. Rust isolates with virulence to Middle American landraces were also generally virulent on Andean material; no truly Middle American-specific isolates were found. Random amplified polymorphic DNA (RAPD) analysis of the rust isolates also distinguished the two groups. Four of the Andean-specific rust isolates formed a distinct group compared to four of the nonspecific isolates. Two of the isolates, one from each of the two virulence groups, had intermediate RAPD banding patterns, suggesting that plasmagomy but not karyogamy occurred between isolates of the two groups.