D. K. Malvick

Characterization of Aphanomyces euteiches strains based on pathogenicity tests and random amplified polymorphic DNA analyses

Genotypic variation among 62 strains of Aphanomyces euteiches, four of A. cochlioides, and a Saprolegnia sp. was investigated using RAPD analysis. Pathogenicity assays on pea, bean, alfalfa, red clover, and sugarbeet were used to determine host preference among the strains of A. euteiches and A. cochlioides. Pathogenicity tests revealed six pathotypes of A. euteiches with host preferences for bean, alfalfa, pea, pea and alfalfa, red clover and alfalfa, and bean and alfalfa. Another group of strains was non-pathogenic on the five plant species. The host of origin tended to be the host on which each strain incited the highest disease severity. A. euteiches did not incite root rot symptoms on sugarbeet, and A. cochlioides was pathogenic only to sugarbeet. RAPD analyses provided a measure of genetic diversity in Aphanomyces. Fifty random decanucleotide primers were screened with five test strains from four hosts representing different pathotypes, and 32 primers amplified DNA fragments from all five strains. Eight primers were chosen for most of this study based on number and polymorphic nature of the bands generated. RAPD assays of 62 strains of A. euteiches with the eight primers yielded 159 polymorphic and no monomorphic, strongly amplified bands. Cluster analyses of RAPD data revealed genotypic differences among three groups of A. euteiches which corresponded to their host of origin and host preference for bean, alfalfa, and red clover/alfalfa. Strains nonpathogenic on all plants tested formed another genotypic group, corroborating results from the pathogenicity assays that indicated this is a discrete group. The bean and non-pathogenic groups were the most distinct. The A. cochlioides and Saprolegnia strains were genotypically distinct from the pathogenic, but not the non-pathogenic strains of A. euteiches. Reproducibility of RAPD assays was confirmed by replicated amplifications and DNA hybridization analysis. Results indicated that A. euteiches is composed of distinct subspecific groups based on genotype and host preference.

Meta-analysis of yield response of hybrid field corn to foliar fungicides in the U.S. corn belt

The use of foliar fungicides on field corn has increased greatly over the past 5 years in the United States in an attempt to increase yields, despite limited evidence that use of the fungicides is consistently profitable. To assess the value of using fungicides in grain corn production, random-effects meta-analyses were performed on results from foliar fungicide experiments conducted during 2002 to 2009 in 14 states across the United States to determine the mean yield response to the fungicides azoxystrobin, pyraclostrobin, propiconazole + trifloxystrobin, and propiconazole + azoxystrobin. For all fungicides, the yield difference between treated and nontreated plots was highly variable among studies. All four fungicides resulted in a significant mean yield increase relative to the nontreated plots (P < 0.05). Mean yield difference was highest for propiconazole + trifloxystrobin (390 kg/ha), followed by propiconazole + azoxystrobin (331 kg/ha) and pyraclostrobin (256 kg/ha), and lowest for azoxystrobin (230 kg/ha). Baseline yield (mean yield in the nontreated plots) had a significant effect on yield for propiconazole + azoxystrobin (P < 0.05), whereas baseline foliar disease severity (mean severity in the nontreated plots) significantly affected the yield response to pyraclostrobin, propiconazole + trifloxystrobin, and propiconazole + azoxystrobin but not to azoxystrobin. Mean yield difference was generally higher in the lowest yield and higher disease severity categories than in the highest yield and lower disease categories. The probability of failing to recover the fungicide application cost (p(loss)) also was estimated for a range of grain corn prices and application costs. At the 10-year average corn grain price of

Biology, Yield loss and Control of Sclerotinia Stem Rot of Soybean

0.12/kg (

Detection and quantification of Phialophora gregata in soybean and soil samples with a quantitative, real-time PCR assay.

2.97/bushel) and application costs of

Traits of soybean-infecting Phytophthora populations from Illinois agricultural fields

40 to 95/ha, p(loss) for disease severity <5% was 0.55 to 0.98 for pyraclostrobin, 0.62 to 0.93 for propiconazole + trifloxystrobin, 0.58 to 0.89 for propiconazole + azoxystrobin, and 0.91 to 0.99 for azoxystrobin. When disease severity was >5%, the corresponding probabilities were 0.36 to 95, 0.25 to 0.69, 0.25 to 0.64, and 0.37 to 0.98 for the four fungicides. In conclusion, the high p(loss) values found in most scenarios suggest that the use of these foliar fungicides is unlikely to be profitable when foliar disease severity is low and yield expectation is high.

Characteristics and Frequency of Aphanomyces euteiches Races 1 and 2 Associated with Alfalfa in the Midwestern United States

Sclerotinia stem rot (also known as white mold) of soybean is a significant yield-limiting problem in the North Central production region. This disease, caused by the fungus Sclerotinia sclerotiorum (Lib.) de Bary, varies in incidence and severity from year to year because of its sensitivity to weather conditions. Losses because of Sclerotinia stem rot can be substantial when environmental conditions and management practices favor high yield potential. Employing a disease management plan based on knowledge of field history and best disease management practices can help reduce losses from Sclerotinia stem rot. An effective disease management plan integrates several management tactics that include cultural practices, varietal resistance, as well as chemical and biological control. Understanding how different environmental variables and management practices influence infection by S . sclerotiorum and disease development are important to optimize disease management and reduce losses. This profile summarizes research-based knowledge of Sclerotinia stem rot, including the disease cycle, the scope of the losses that can occur because of this disease, how to identify both the pathogen S . sclerotiorum and the disease, and current management recommendations.

Genotypic and pathogenic diversity among pea-infecting strains of Aphanomyces euteiches from the central and western United States.

Brown stem rot of soybean, caused by the soilborne fungus Phialophora gregata, is a common and widespread disease of soybean (Glycine max) in the midwestern United States. This pathogen is challenging to study due to a long latent period and slow growth. A TaqMan probe-based quantitative, real-time polymerase chain reaction (qPCR) assay was developed for sensitive and specific detection and quantification of genotypes A and B of P. gregata in plant and soil samples. It is sensitive with detection limits of 50 fg of pure genomic DNA, 100 copies of the target DNA sequence, and approximately 400 conidia. The qPCR assay is approximately 1,000 times more sensitive in detecting DNA and conidia of P. gregata, and is more rapid and less sensitive to PCR inhibitors from soybean stems than a standard PCR (sPCR) assay. Using this single-step qPCR assay, low levels of infection were detected in soybean stems at least 1 to 2 weeks prior to symptom development and before P. gregata was detected with sPCR. This assay also was used to detect the pathogen in field-grown plants and in naturally infested field soils. This new qPCR assay is a powerful tool for rapid, specific, and sensitive detection, diagnosis, and quantification of P. gregata in plants and soil, and for advancing studies of the ecology of P. gregata and its interactions with host plants.

Variation in pathogenicity and genotype among single-zoospore strains of Aphanomyces euteiches

Phytophthora rot caused by Phytophthora sojae is a common and significant disease of soybean (Glycine max) in Illinois and throughout the Midwestern United States. The pathogenic characteristics of P. sojae populations in several Midwestern states have been reported recently, but pathogenicity and fungicide sensitivity traits of populations in Illinois were poorly understood. Isolates (n = 121) of soybean-infecting Phytophthora spp. were baited using susceptible cv. Sloan seedlings from soybean field soils with a history of seedling diseases in 24 counties across Illinois. The pathotype and race of isolates of P. sojae were characterized using 11 differential soybean cultivars in greenhouse tests using a hypocotyl inoculation method. Sensitivity to the fungicidal compounds metalaxyl and mefenoxam was tested with 63 isolates in vitro. Most (96%) of the Phytophthora isolates sampled from Illinois soybean fields were P. sojae, but 4% were an unidentified Phytophthora sp. as determined by phenotypic and genotypic traits. We present a preliminary description of another Phytophthora sp. from soybean fields in a restricted region of Illinois that is pathogenic and capable of killing soybean. Based on eight Rps gene differentials (Rps1a, 1b, 1c, 1d, 1k, 3a, 6, and 7 ), 22 virulence pathotypes of P. sojae were identified and 88% of all isolates were characterized to a defined race. The four most common races, which were 58% of all isolates, were races 1 (21%), 4 (15%), 33 (12%), and 28 (10%). Based on 11 differentials, (those noted above and Rps 2, 4, and 5), 31 virulence pathotypes were identified. The mean virulence complexities, which are the number of susceptible interactions on the sets of 8 and 11 Rps gene differentials, were 3.3 and 3.7, respectively. All isolates tested were sensitive to Apron XL, Allegiance, technical grade mefenoxam, and technical grade metalaxyl at 1.0 μg a.i./ml. The population of P. sojae is diverse and composed of multiple pathotypes and races in Illinois, and the results suggest that pathogen virulence partially explains poor performance of Phytophthora-resistant cultivars in many Illinois soybean fields.

Identification of Pisum sativum Germ Plasm with Resistance to Root Rot Caused by Multiple Strains of Aphanomyces euteiches

Aphanomyces root rot of alfalfa, caused by Aphanomyces euteiches, kills seedlings and causes decline of established plants in slowly drained soils. Two races of A. euteiches that are pathogenic to alfalfa have been identified. Despite the contribution of race 1 resistance to establishment and yield of alfalfa, race 1-resistant alfalfa cultivars perform poorly in some fields infested with A. euteiches. Many isolates of A. euteiches obtained from the soils of problematic fields are of a race 2 phenotype. The purpose of this study was to determine distribution, frequency, and pathogenic and genotypic characteristics of race 1 (R1) and race 2 (R2) isolates from 21 fields: 13 in Wisconsin, 7 in Minnesota, and 1 in Kentucky. A. euteiches was successfully isolated from the soil of 16 of the 21 fields; 405 isolates were obtained from Wisconsin, 4 from Minnesota, and 48 from Kentucky. Pathogenicity and race phenotype of isolates were characterized on Saranac (susceptible to R1 and R2 isolates) and WAPH-1 (resistant to R1 and susceptible to R2 isolates) alfalfa populations. One Wisconsin field with no recent history of alfalfa production had a high frequency (51%) of R2 isolates, and 43% of all isolates were R2 from fields with a history of alfalfa production. In a location that was planted continuously to pea for 30 years, 27% of the isolates were R2. Random amplified polymorphic DNA (RAPD) analysis of three R1 and three R2 isolates with eight primers generated 43 total polymorphic bands; however, none of the bands were uniquely associated with race phenotype. Cluster analysis based on RAPD bands revealed no consistent genotypic distinctions between R1 and R2 isolates of A. euteiches. Evaluation of eight commercial alfalfa cultivars for resistance to two R1 and two R2 isolates demonstrated that most are susceptible to R2 isolates; however, those selected for R2 resistance express resistance to R2 isolates. The results suggest that R2 isolates represent a widespread risk to alfalfa cultivars having resistance only to R1 isolates in fields with varied cropping histories.

Corn yield loss estimates due to diseases in the United States and Ontario, Canada from 2012 to 2015.

ABSTRACT Pathogenic and genotypic variability among four populations of Aphanomyces euteiches from individual fields in Minnesota, Wisconsin, and Oregon were investigated using pathogenicity and randomly amplified polymorphic DNA (RAPD) analyses. About 50 strains were isolated from each of two pea fields in Minnesota, and 11 and 6 strains from pea fields in Wisconsin and Oregon, respectively, using pea (Pisum sativum) as a baiting host. Pathogenic variability and host range were evaluated in greenhouse studies with five pea lines or cultivars having different levels of resistance to Aphanomyces root rot and one cultivar each of alfalfa and snap bean. All strains were pathogenic on one or more pea cultivars, and 18 and 14% were pathogenic on alfalfa and bean, respectively. Disease severity incited by different strains varied significantly on individual pea cultivars and on all hosts combined. The percentage of strains pathogenic on different hosts varied among locations. Genotypic variation among all 114 strains was evaluated with RAPD analysis. Ten decanucleotide primers detected 92 polymorphic bands. Cluster and principal coordinates analysis revealed one large group containing 102 of the 114 strains from all locations. Two closely related minor groups of strains (12 strains) were genotypically distinct, with about 55% similarity to the main group of 102 strains. The strains in the minor groups were all isolated from the Minnesota locations and were pathogenic on two disease-resistant pea breeding lines (MN313 and MN314). Estimates of genetic diversity based on RAPD analysis ranged from 0.24 to 0.33 within populations to 0.35 among all strains from all populations. A. euteiches populations were genotypically and phenotypically variable, but no distinct genotypic differences were identified among populations from the four isolated locations.