Comparison of baiting techniques for recovering Phytophthora sojae from soybean fields in Iowa.

Understanding the diversity of a pathogen is important for developing disease management recommendations. In the Phytophthora root and stem rot (PRSR)-soybean pathosystem, Phytophthora sojae is characterized into pathotypes based on the ability of the pathogen to cause disease on soybean genotypes that each contain a different resistance Rps gene. To understand the diversity of P. sojae in an area, isolates of the pathogen may be recovered by baiting soil samples. In this research, we compared two commonly used methods for baiting P. sojae: the leaf disc method and the seedling method. More isolates of P. sojae were recovered using the leaf disc baiting method (P<0.01), and more pathotypes were detected within this population (P<0.01). Mean complexity of the two populations and the Simple, Gleason, and Shannon diversity indices also differed (P<0.01). However, the percent of isolates that caused disease on each Rps gene did not differ between methods. Thus, either method could be used to characterize pathotypes of P. sojae present in an area to provide data to soybean breeders and agronomists for deployment of Rps genes in soybean cultivars.

Pathotype Complexity and Genetic Characterization of Phytophthora sojae Populations in Illinois, Indiana, Kentucky, and Ohio

Phytophthora sojae, the causal agent of Phytophthora root and stem rot of soybean, has been managed with single Rps genes since the 1960’s, but has subsequently adapted to many of these resistance genes, rendering them ineffective. The objective of this study was to examine the pathotype and genetic diversity of P. sojae from soil samples across Illinois, Indiana, Kentucky, and Ohio by assessing which Rps gene(s) were still effective and identifying possible population clusters. There were 218 pathotypes identified from 473 P. sojae isolates with an average of 6.7 out of 15 differential soybean lines exhibiting a susceptible response for each isolate. Genetic characterization of 103 P. sojae isolates from across Illinois, Indiana, Kentucky, and Ohio with 19 simple sequence repeat markers identified 92 multilocus genotypes. There was a moderate level of population differentiation among these four states, with pairwise FST values ranging from 0.026 to 0.246. There was also moderate to high levels of differentiation between fields, with pairwise FST values ranging from 0.071 to 0.537. Additionally, cluster analysis detected the presence of P. sojae population structure across neighboring states. The level of pathotype and genetic diversity, in addition to the identification of population clusters, supports the hypothesis of occasional outcrossing events that allow for an increase in diversity and the potential to select for a loss in avirulence to specific resistance genes within regions. The trend of suspected gene flow among neighboring fields is expected to be an ongoing issue with current agricultural practices.

Comparison of Phytophthora sojae populations in Iowa and Nebraska to identify effective Rps genes for Phytophthora stem and root rot management

Phytophthora stem and root rot (PSRR) of soybean, caused by the oomycete Phytophthora sojae, is prevalent in Iowa and Nebraska. Reducing losses to PSRR primarily relies on growing cultivars with specific resistance (Rps) genes. Predominant genes used in commercial soybean cultivars include Rps 1a, Rps 1c, Rps 1k, and Rps 3a. Knowing which Rps gene to deploy depends on knowledge of which genes are effective against the pathogen. From 2016 to 2018, 326 isolates of P. sojae from were recovered from fields in Iowa and Nebraska and classified into pathotypes based on their virulence on 15 soybean genotypes. A total of 15 and 10 pathotypes were identified in Iowa and Nebraska, respectively. Almost all isolates were virulent on Rps 1a, while over 70% of isolates were virulent on Rps 1c and Rps 1k. Only 2.3% of isolates from Iowa were virulent on Rps 3a. Among commercial soybean cultivars tested in the Illinois Soybean Variety trials from 2010 to 2020, Rps 1c was always the most frequently reported gene followed by Rps 1k. In contrast, Rps 1a and Rps 3a were present in less than 10% and less than 5 % of the cultivars tested, respectively. Since many of the P. sojae isolates in our study were virulent on Rps 1a, Rps 1c, and Rps 1k, soybean cultivars with these genes are unlikely to provide protection against PSRR unless they have a high level of partial resistance.

Common Phytophthora sojae pathotypes occurring in South Dakota

Phytophthora root and stem rot, caused by Phytophthora sojae, is an important disease of soybean (Glycine max L.) in South Dakota. Because P. sojae populations are highly diverse and resistance genes deployed in commercial soybean varieties often fail to manage the disease, this study was initiated to determine P. sojae pathotype distribution in South Dakota. A total of 216 P. sojae isolates were baited from soil collected from 422 soybean fields in South Dakota in 2013-2015 and 2017. The pathotype of each isolate was determined by inoculating 10 seedlings of 13 standard soybean P. sojae differential lines using the hypocotyl inoculation technique. Of the 216 pathotyped isolates, 48 unique pathotypes were identified. The virulence complexity of isolates ranged from virulence on one Rps gene (Rps7) to virulence on 13 Rps genes and mean complexity was 5.2. Harosoy (Rps7), Harlon (Rps1a), Williams 79 (Rps 1c), William 82 (Rps1k), Harosoy 13XX (Rps1b), were susceptible to 98, 80, 78, 73, 72% of the isolates, respectively. These results highlight the highly diverse P. sojae pathotypes in South Dakota and the likely Rps genes to fail in commercial soybean varieties

Phytophthora sojae pathotype distribution and fungicide sensitivity in Michigan

Identifying the pathotype structure of a Phytophthora sojae population is crucial for the effective management of Phytophthora stem and root rot of soybean (PRR). P. sojae has been successfully managed with major resistance genes, partial resistance, and fungicide seed treatments. However, prolonged use of resistance genes or fungicides can cause pathogen populations to adapt over time, rendering resistance genes or fungicides ineffective. A statewide survey was conducted to characterize the current pathotype structure and fungicide sensitivity of P. sojae within Michigan. Soil samples were collected from 69 fields with a history of PRR and fields having consistent plant stand establishment issues. Eighty-three isolates of P. sojae were obtained, and hypocotyl inoculations were performed on 14 differential soybean cultivars, all of which carry a single Rps gene or no resistance gene. The survey identified a loss of effectiveness of Rps genes 1b, 1k, 3b and 6, compared to a previous survey conducted in Michigan from 1993-1997. Three effective resistance genes were identified for P. sojae management in Michigan; Rps 3a, 3c, and 4. Additionally, the effective concentration of common seed treatment fungicides to inhibit mycelial growth by 50% (EC50) was determined. No P. sojae isolates were insensitive to the tested chemistries with mean EC50 values of 2.60x10-2 µg/ml for ethaboxam, 3.03x10-2 µg/ml for mefenoxam, 2.88x10-4 µg/ml for oxathiapiprolin, and 5.08x10-2 µg/ml for pyraclostrobin. Results suggest that while there has been a significant shift in Rps gene effectiveness, seed treatments are still effective for early season management of this disease.

Effect of Resistance and Ethaboxam Seed Treatment on the Management of Phytophthora Root Rot in Illinois and Iowa

Phytophthora root and stem rot (PRR) is a limiting factor for soybean production. Seed treatments are used for early-season management, but efficacy can depend on seed selection and the local environment. Ethaboxam is a new fungicide commercially available as a seed treatment to control oomycetes. Field experiments were established in Illinois and Iowa in 2017 and 2018 to evaluate the effect of ethaboxam + metalaxyl on PRR. Experiments included soybean lines with no resistance gene, Rps1c or Rps1k, and different levels of partial resistance. Seed treatments increased soybean stands in all locations and years. Significant yield effects were observed only in two locations that were inoculated with Phytophthora spp. Groups of soybean lines with the same Rps gene responded differently in each location, showing how Rps gene usefulness depends on the field. A comparison of the effect of seed treatment on lines with different levels of partial resistance showed that partial resistance alone cannot always protect against stand losses. Soybean lines with high levels of partial resistance had consistently higher yields than those with low levels of partial resistance across Illinois locations. These results show that ethaboxam seed treatment can protect early-season stands and that selection of cultivars with high levels of partial resistance is important for PRR management.

Pathotype Diversity of Phytophthora sojae in Eleven States in the United States

Pathotype diversity of Phytophthora sojae was assessed in 11 states in the United States during 2012 and 2013. Isolates of P. sojae were recovered from 202 fields, either from soil samples using a soybean seedling bioassay or by isolation from symptomatic plants. Each isolate was inoculated directly onto 12 soybean differentials; no Rps gene or Rps 1a, 1b, 1c, 1k, 3a, 3b, 3c, 4, 6, 7, or 8. There were 213 unique virulence pathotypes identified among the 873 isolates collected. None of the Rps genes were effective against all the isolates collected but Rps6 and Rps8 were effective against the majority of isolates collected in the northern regions of the sampled area. Virulence toward Rps1a, 1b, 1c, and 1k ranged from 36 to 100% of isolates collected in each state, while virulence to Rps6 and Rps8 was less than 36 and 10%, respectively. Depending on the state, the effectiveness of Rps3a ranged from totally effective to susceptible to more than 40% of the isolates. Pathotype complexity has increased in populations of P. sojae in the United States, emphasizing the increasing importance of stacked Rps genes in combination with high partial resistance as a means of limiting losses to P. sojae.

A Method for Combining Isolates of Phytophthora sojae to Screen for Novel Sources of Resistance to Phytophthora Stem and Root Rot in Soybean

Soybean cultivars with specific single resistance genes (Rps) are grown to reduce yield loss due to Phytophthora stem and root rot caused by the oomycete pathogen Phytophthora sojae. To identify novel Rps loci, soybean lines are often screened several times, each time with an isolate of P. sojae that differs in virulence on various Rps genes. The goal of this study was to determine whether several isolates of P. sojae that differ in virulence on Rps genes could be combined into a single source of inoculum and used to screen soybean lines for novel Rps genes. A set of 14 soybean differential lines, each carrying a specific Rps gene, was inoculated with three isolates of P. sojae, which differed in virulence on 6 to 10 Rps genes, individually or in a 1:1:1 mixture. Inoculum containing the 1:1:1 mixture of isolates was virulent on 13 Rps genes. The mixed-inoculum method was used to screen 1,019 soybean accessions in a blind assay for novel sources of resistance. In all, 17% of Glycine max accessions and 11% of G. soja accessions were resistant (≤30% dead plants), suggesting that these accessions may carry a novel Rps gene or genes. Advantages of combining isolates into a single source of inoculum include reduced cost, ability to screen soybean germplasm with inoculum virulent on all known Rps genes, and ease of identifying novel sources of resistance. This study is a precursor to identifying novel sources of resistance to P. sojae in soybean using RXLR effectors.

Population Structure Among and Within Iowa, Missouri, Ohio, and South Dakota Populations of Phytophthora sojae

Phytophthora root and stem rot, caused by Phytophthora sojae, is an economically important disease of soybean throughout the Midwestern United States. This disease has been successfully managed with resistance (Rps) genes; however, pathogen populations throughout the Midwest have developed virulence to many Rps genes, including those that have not been deployed. To gain a better understanding of the processes that influence P. sojae evolution, the population genetic structure was compared among populations using one isolate collected from 17, 33, and 20 fields in Iowa, Ohio, and South Dakota, respectively, as well as multiple isolates from individual fields in Iowa, Ohio, and Missouri. Genotypic diversity was measured using 21 polymorphic microsatellite (simple-sequence repeat) markers. and pathotype diversity using 15 soybean differentials. For all but three of the populations with low sample size, there was a high level of pathotype diversity and a low to moderate level of genotypic diversity among the populations for both comparisons between states and within-field variation. None of the Rps-gene differentials were resistant to all of the isolates. There were 103 unique multilocus genotypes identified in this study and only 2 were identified from the same field. Although no clones were identified in more than one field, pairwise FST indicated that some gene flow within neighboring fields does occur but not across the region, including fields from neighboring states. These results suggest that there is a strong probability that each state may have their own or several regional populations, as well as provide further evidence of high diversity within this homothallic pathogen which may be due, in part, to limited gene flow, mutation, or outcrossing, and this likely affects the success of deployment of resistance.