scholarly journals Pathogenic Variation of Phakopsora pachyrhizi Isolates on Soybean in the United States from 2006 to 2009

Plant Disease ◽  
2012 ◽  
Vol 96 (1) ◽  
pp. 75-81 ◽  
Author(s):  
M. Twizeyimana ◽  
G. L. Hartman

The introduction of Phakopsora pachyrhizi, the cause of soybean rust, into the United States is a classic case of a pathogen introduction that became established in a new geographical region overwintering on a perennial host (kudzu, Pueraria lobata). The objective of our study was to classify the pathogenic variation of P. pachyrhizi isolates collected in the United States, and to determine the spatial and temporal associations. In total, 72 isolates of P. pachyrhizi collected from infected kudzu and soybean leaves in the United States were purified, then established and increased on detached soybean leaves. These isolates were tested for virulence and aggressiveness on a differential set of soybean genotypes that included six genotypes with known resistance genes (Rpp), one resistant genotype without any known characterized resistance gene, and a susceptible genotype. Three pathotypes were identified among the 72 U.S. P. pachyrhizi isolates based on the virulence of these isolates on the genotypes in the differential set. Six aggressiveness groups were established based on sporulating-uredinia production recorded for each isolate on each soybean genotype. All three pathotypes and all six aggressiveness groups were found in isolates collected from the southern region and from both hosts (kudzu or soybean) in 2008. Shannon's index based on the number of pathotypes indicated that isolates from the South region were more diverse (H = 0.83) compared with the isolates collected in other regions. This study establishes a baseline of pathogenic variation of P. pachyrhizi in the United States that can be further compared with variation reported in other regions of the world and in future studies that monitor P. pachyrhizi virulence in association to deployment of rust resistance genes.

Plant Disease ◽  
2013 ◽  
Vol 97 (10) ◽  
pp. 1379-1379 ◽  
Author(s):  
C. Paul ◽  
G. L. Hartman ◽  
J. J. Marois ◽  
D. L. Wright ◽  
D. R. Walker

During the years following the first detection of soybean rust, caused by Phakopsora pachyrhizi Syd., in the continental United States in November, 2004, soybean (Glycine max [L.] Merr.) genotypes with the Rpp1 or Rpp6 resistance genes exhibited high levels of resistance there (1,2,3). When challenged with 72 different American isolates collected between 2006 and 2009, PI 200492 (source of Rpp1) produced no sporulating lesions (2). In 2011 and 2012, however, field populations of P. pachyrhizi from Gadsden County, FL, caused higher rust severity on plants with Rpp1 or Rpp6 than in previous years. To assess aggressiveness, sporulation ratings were made using a 1 to 5 scale (no sporulation to profuse sporulation) on leaflets collected from field plants at or near the R6 (full seed) stage of development. A dissecting microscope was used to examine 3 replications of 5 leaflets each in 2009 or 2 replications of 10 leaflets each in 2012. The sporulation ratings increased on PI 200492 (from 1.1 ± 0.1 in 2009 to 4.1 ± 0.4 in 2012), PI 567102B (Rpp6; from 1.1 ± 0.1 in 2009 to 2.4 ± 0.2 in 2012), and L85-2378, a ‘Williams 82’ isoline carrying the Rpp1 gene (from 1.0 ± 0 in 2009 to 4.0 ± 0.3 in 2012). The mean 2009 and 2012 sporulation ratings for susceptible control Williams 82 were 5.0 ± 0 and 4.2 ± 0.1, respectively. Single-uredinium-derived isolates were purified from bulk isolates collected from field plots in 2009 (FL-Q09-1), 2011 (FL-Q11-1), and 2012 (FL-Q12-1). Greenhouse and detached leaflet assays were then used to test the virulence of these isolates under controlled conditions. Detached leaflets from 3-week-old seedlings of Williams 82, PI 200492, PI 567102B, and L85-2378 were inoculated by pipetting 15-μl drops of a 30 to 40 urediniospore μl–1 suspension onto the abaxial side of 3 to 4 leaflets per genotype, which were then sealed in Petri plates and incubated in a growth chamber at 20 to 22°C. Plates were kept in the dark for 12 h following inoculation. For the greenhouse assay, the first trifoliolate leaves of at least 3 seedlings were each sprayed with 1.5 ml of a 40 urediniospore μl–1 suspension and incubated 24 h at 22 to 24°C in a dark mist chamber. The plants were then maintained at 22 to 24°C and 76 to 86% relative humidity in a greenhouse with 10 h of daylight on average. Two weeks after inoculation with FL-Q11-1 or FL-Q12-1, all of the genotypes had developed TAN lesions with abundant sporulation, indicating susceptibility. On leaves inoculated with FL-Q09-1, however, no visible reaction was observed on PI 200492, and PI 567102B developed reddish-brown (RB) lesions associated with incomplete resistance. Although the lesions on Rpp1 and Rpp6 greenhouse seedlings inoculated with the FL-Q11-1 and FL-Q12-1 isolates were slightly darker than those that developed on Williams 82 plants or on detached leaflets, the profuse sporulation that is characteristic of the TAN infection type was observed. The higher virulence of the 2011 and 2012 Florida isolates on two soybean genotypes with Rpp1 and one with Rpp6 confirmed the presence of a P. pachyrhizi pathotype in north-central Florida that is more virulent against these genes than earlier populations from the southeastern United States. References: (1) S. Li. Crop Sci. 49:887, 2009. (2) Twizeyimana and Hartman. Plant Dis. 96:75, 2012. (3) Walker et al. Crop Sci. 51:678, 2011.


Plant Disease ◽  
2015 ◽  
Vol 99 (8) ◽  
pp. 1059-1069 ◽  
Author(s):  
C. Paul ◽  
R. D. Frederick ◽  
C. B. Hill ◽  
G. L. Hartman ◽  
D. R. Walker

A major constraint in breeding for resistance to soybean rust has been the virulence diversity in Phakopsora pachyrhizi populations. In greenhouse experiments, reactions of 18 soybean genotypes to 24 U.S. isolates from 2007 and 2008 and 4 foreign isolates were compared. Reactions of four differentials (Rpp1 to Rpp4) to these U.S. isolates were also compared with reactions to nine foreign isolates and three U.S. isolates from 2004. Principal component analysis (PCA) of the reaction types grouped the U.S. isolates into a single virulence group, whereas each of the foreign isolates had a unique virulence pattern. In another experiment, reactions of 11 differentials to the 24 U.S. isolates were compared and significant interactions (P < 0.001) were found between the isolates and host genotypes for rust severity and uredinia densities. PCA of these two measures of disease placed the 24 isolates into seven or six aggressiveness groups, respectively. In a third experiment, evaluation of 20 soybean genotypes for resistance to the previously established aggressive groups identified 10 genotypes resistant to isolates representing most of the groups. This study confirmed the pathogenic diversity in P. pachyrhizi populations and identified soybean germplasm with resistance to representative U.S. isolates that can be used in breeding.


Plant Disease ◽  
2014 ◽  
Vol 98 (9) ◽  
pp. 1261-1266 ◽  
Author(s):  
M. Twizeyimana ◽  
P. S. Ojiambo ◽  
R. Bandyopadhyay ◽  
G. L. Hartman

Soybean rust, caused by Phakopsora pachyrhizi, is one of the most important foliar diseases of soybean worldwide. The soybean–P. pachyrhizi interaction is often complex due to genetic variability in host and pathogen genotypes. In a compatible reaction, soybean genotypes produce tan-colored lesions, whereas in an incompatible reaction soybean genotypes produce an immune response (complete resistance) or reddish-brown lesions (incomplete resistance). In this study, in total, 116 and 72 isolates of P. pachyrhizi from Nigeria and the United States, respectively, were compared based on six quantitative traits to assess their aggressiveness on two soybean genotypes. All isolates produced reddish-brown lesions on plant introduction (PI) 462312 and tan lesions on TGx 1485-1D. The number of days after inoculation to first appearance of lesions, uredinia, and sporulation, along with the number of lesions and sporulating uredinia per square centimeter of leaf tissue, and the number of uredinia per lesion, were significantly (P < 0.001) different between the two soybean genotypes for all isolates from each country. The number of days to first appearance of lesions, uredinia, and sporulation were greater on PI 462312 than on TGx 1485-1D for all the test isolates. Similarly, the number of lesions and sporulating uredinia per square centimeter, and the number of uredinia per lesion were lower on PI 462312 than on TGx 1485-1D. For both soybean genotypes, the number of sporulating uredinia per square centimeter significantly (P = 0.0001) increased with an increase in the number of lesions per square centimeter. Although the slope of the regression of sporulating uredinia on number of lesions was greater (P < 0.0001) when TGx 1485-1D was inoculated with Nigerian isolates compared with U.S. isolates, slopes of the regression lines did not differ significantly (P > 0.0675) when PI 46312 was inoculated with Nigerian or U.S. isolates. This is the first study that used a large number of isolates from two continents to assess aggressiveness of P. pachyrhizi using multiple traits in soybean genotypes with contrasting types of disease reaction.


Plant Disease ◽  
2006 ◽  
Vol 90 (1) ◽  
pp. 112-112 ◽  
Author(s):  
J. M. Mullen ◽  
E. J. Sikora ◽  
J. M. McKemy ◽  
M. E. Palm ◽  
L. Levy ◽  
...  

On November 4, 2004, soybean leaves (Glycine max (L.) Merr) were submitted to the Auburn University Plant Diagnostic Lab by a State Department of Agriculture and Industries Inspector. Samples were collected from an 80-ha field of soybean plants in a late-reproductive-growth stage in Mobile County, Alabama. Under microscopic examination, leaves showed rust pustules in advanced stages of development with urediniospores and sori characteristic of Phakopsora spp. Uredinia were ostiolate in small, brown, angular leaf spots (2 to 3 mm) on lower leaf surfaces. Urediniospores were pale yellow-to-white, globose or ovate, 20 to 40 × 15 to 25 μm. In a subsequent visit to the field, symptoms and signs of the rust disease were observed on plants bordering the edge of the field since the majority of plants were senescent. Tan lesions on lower leaf surfaces contained small pustules surrounded by a small zone of slightly discolored necrotic tissue. Masses of tan spores covered the lower leaf surface pustules. Leaves were mailed overnight to the USDA National Identification Services (Mycology) Laboratory in Beltsville, MD. The fungal structures were confirmed to be a Phakopsora sp., and the sample was forwarded to the USDA National Plant Germplasm and Biotechnology Laboratory in Beltsville, MD. DNA was extracted from leaf pieces containing sori using the Qiagen DNeasy Plant Mini kit (Qiagen, Valencia, CA). Phakopsora pachyrhizi was detected using a real-time polymerase chain reaction (PCR) protocol (1) performed in a Cepheid SmartCycler (Sunnyvale, CA). The PCR master mix was modified to include OmniMix beads (Cepheid). The field and microscopic suspect diagnosis of P. pachyrhizi was confirmed officially by APHIS on November 18, 2004. This was the fourth USDA official confirmation of Asian soybean rust in the continental United States during 2004, and to our knowledge, this is the first report of the disease in Alabama. This report helps confirm that early occurrences of Asian soybean rust in the United States were present in other areas in addition to the first reported finding in Louisiana (2). References: (1) R. D. Frederick et al. Phytopathology 92:217, 2002. (2) R. W. Schneider et al. Plant Dis. 89:774, 2005.


Plant Disease ◽  
2011 ◽  
Vol 95 (8) ◽  
pp. 1034-1034
Author(s):  
M. A. Delaney ◽  
E. J. Sikora ◽  
D. P. Delaney ◽  
M. E. Palm ◽  
J. Roscoe ◽  
...  

Soybean rust, caused by the fungus Phakopsora pachyrhizi, was detected on jicama (Pachyrhizus erosus L. Urban) for the first time in the United States in November 2009. The pathogen was observed on leaves of a single, potted jicama plant grown outdoors in a residential area and on leaves of all plants in a 12-m2 demonstration plot located at the Auburn University Teaching Garden in Auburn, AL. Symptoms on the upper leaf surfaces were isolated chlorotic areas near the leaf edges in the lower part of the canopy. The abaxial surface was first observed to exhibit brown lesions and subsequently produced volcano-shaped uredinia. These symptoms are consistent with a rust previously described on jicama in Mexico (1). Representative symptomatic plant tissue was sent to the USDA National Identification Services (Mycology) Laboratory in Beltsville, MD for diagnostic confirmation at both the Urbana, IL lab and the USDA National Plant Germplasm and Biotechnology Laboratory for DNA testing. From an infected leaf, samples of approximately 5 mm2 were excised from a microscopically observed rust lesion and an apparently noninfected area. Total DNA was purified with the FastDNA Spin Kit (MP Biomedicals, Solon, OH) followed by the E.Z.N.A. MicroElute DNA Clean-Up Kit (Omega Bio-tek, Inc, Doraville, GA) per manufacturer's instructions. Detection of P. pachyrhizi and P. meibomiae DNA was achieved by quantitative PCR using the method of Frederick et al. (2) and a DNA standard of previously prepared P. pachyrhizi spores. The observed rust pustule was found to contain P. pachyrhizi DNA in excess of 28,000 genomes, while no P. pachyrhizi DNA was observed from the asymptomatic sample. Both samples were negative for P. meibomiae. The fungal structures present were confirmed to be Phakopsora spp. DNA was extracted from sori aseptically removed from leaves with a Qiagen (Valencia, CA) DNeasy Plant Mini Kit and amplified with primers Ppa1 and NL4. The resulting partial ITS2 and 28S ribosomal RNA sequences were 100% identical to GenBank entry DQ354537 P. pachyrhizi internal transcribed spacer 2 and 28S ribosomal RNA gene, partial sequence. Sequences from jicama from Alabama were deposited in GenBank. Voucher specimens were deposited in the USDA Agricultural Research Service, National Fungus Collection (BPI). To our knowledge, this is the first report of the disease on jicama in the United States. References: (1) A. Cárcamo Rodriguez et al. Plant Dis. 90:1260, 2006. (2) R. D. Frederick et al. Phytopathology 92:217, 2002.


2014 ◽  
Vol 1 (1) ◽  
Author(s):  
Mateus Rodrigues Pereira ◽  
Bianca Castro Gouvêa ◽  
Francismar Corrêa Marcelino-Guimarães ◽  
Humberto Josué de Oliveira Ramos ◽  
Maurilio Alves Moreira ◽  
...  

AbstractAsian soybean rust (ASR), which is incited by the fungus Phakopsora pachyrhizi, is considered one of the most aggressive diseases to the soybean culture. There are no commercial cultivars immune to the pathogen and the control measure currently used is the application of fungicides that harms the environment and increases production costs. For a better understanding of the host’s response to the pathogen at the molecular level, two soybean genotypes were analyzed (PI561356, resistant to ASR and Embrapa 48, susceptible) at 72 hours and 192 hours after inoculation with spores of P. pachyrhizi. Leaf protein profiles of the plants were compared by two-dimensional electrophoresis associated with mass spectrometry (MS). Twenty-two protein spots presented different levels when the two treatments were compared (inoculated vs. non-inoculated). From those, twelve proteins were identified by MS analysis. Some of them are involved in metabolic pathways related to plant defense against pathogens, as in the case of carbonic anhydrase, 1-deoxy-D-xylulose- 5-phosphate reductoisomerase, fructose-bisphosphate aldolase and glutamine synthetase. The possible biochemical-physiological meanings of our findings are discussed.


Plant Disease ◽  
2005 ◽  
Vol 89 (6) ◽  
pp. 678-682 ◽  
Author(s):  
S. Pivonia ◽  
X. B. Yang ◽  
Z. Pan

This article assesses the epidemic potential of soybean rust (Phakopsora pachyrhizi) in the United States. In the assessment, there are three critical components of uncertainty: (i) suitability of climate conditions in production areas for soybean rust epidemics; (ii) likelihood of establishment of the fungus in North America; and (iii) the seasonal dispersal potential of the pathogen from overwintering regions to major soybean production regions. Assessments on the first and second components suggest soybean rust epidemics are likely in the United States, and the certainty of the third component is yet to be determined. Comparison of epidemiological factors for soybean rust in soybean production regions between China and the United States shows a complicated picture with the United States having factors that both increase and decrease risk. Future investigation of risk components—incipience in the field and long-distance dispersal—is needed.


2009 ◽  
Vol 99 (12) ◽  
pp. 1412-1420 ◽  
Author(s):  
Anatoliy V. Lygin ◽  
Shuxian Li ◽  
Ramya Vittal ◽  
Jack M. Widholm ◽  
Glen L. Hartman ◽  
...  

Understanding the metabolic responses of the plant to a devastating foliar disease, soybean rust, caused by Phakopsora pachyrhizi, will assist in development of cultivars resistant to soybean rust. In this study, differences in phenolic metabolism were analyzed between inoculated and noninoculated plants using two susceptible and three resistant soybean genotypes with known resistance genes. Rust infection resulted in increased accumulation of isoflavonoids and flavonoids in leaves of all soybean genotypes tested. Although the soybean phytoalexin glyceollin was not detected in leaves of uninfected plants, accumulation of this compound at marked levels occurred in rust-infected leaves, being substantially higher in genotypes with a red-brown resistant reaction. In addition, there was inhibition of P. pachyrhizi spore germination by glyceollin, formononetin, quercetin, and kaempferol. However, there was no correlation between concentrations of flavonoids quercetin and kaempferol and rust-induced isoflavonoid formononetin in soybean leaves and rust resistance. Lignin synthesis also increased in all inoculated soybean genotypes whereas there was no significant difference in all noninoculated soybean genotypes. Cell wall lignification was markedly higher in inoculated resistant lines compared with inoculated susceptible lines, indicating a possible protective role of lignin in rust infection development.


Plant Disease ◽  
2006 ◽  
Vol 90 (7) ◽  
pp. 972-972 ◽  
Author(s):  
L. E. Sconyers ◽  
R. C. Kemerait ◽  
J. H. Brock ◽  
R. D. Gitaitis ◽  
F. H. Sanders ◽  
...  

Phakopsora pachyrhizi Syd. & P. Syd., which causes Asian soybean rust (SBR), was observed on Florida beggarweed, Desmodium tortuosum (Sw) DC., in Attapulgus, GA during late October and early November 2005. Tan to brown lesions (<1.0 mm in diameter) consistent with symptoms of SBR (2) were observed on older leaves of several plants collected near an SBR-infected soybean trial. Dissection (40 to 60×) and compound microscopy (×200 to 400) revealed conical pustules and ellipsoid, echinulate urediniospores (average size 15 × 20 μm) on the abaxial leaf surface. Polymerase chain reaction (PCR) (primers Ppm1 and Ppa2) (1) was conducted on four samples to confirm identification of P. pachyrhizi or P. meibomiae. Three were positive for P. pachyrhizi, and one was negative for both species. Using morphology and real-time PCR, SBR was confirmed as P. pachyrhizi by the USDA/APHIS in Beltsville, MD. Six noninfected Florida beggarweed plants were transplanted to pots during December 2005 and grown at 22 to 24°C in a greenhouse. On 11 January 2006, a water suspension of urediniospores collected from SBR-infected soybeans (1 × 105 spores per ml) was spray inoculated on all leaves to almost runoff and incubated for 48 h in a plastic humidity chamber. Lesions, pustules, and urediniospores consistent with SBR (2) were observed on 3 February 2006. A PCR assay was conducted on six samples from the infected greenhouse plants and all were positive for P. pachyrhizi. Florida beggarweed is widespread in the southern United States and may serve as an additional overwintering source for P. pachyrhizi and a potential inoculum source for the soybean crop. References: (1) R. D. Fredrick et al. Phytopathology 92:217, 2002. (2) J. B. Sinclair and G. L. Hartman. Soybean rust. Pages 25–26 in: Compendium of Soybean Diseases. 4th ed. G. L. Hartman et al., eds. The American Phytopathological Society, St. Paul, MN, 1999.


Plant Disease ◽  
2015 ◽  
Vol 99 (11) ◽  
pp. 1604-1609 ◽  
Author(s):  
Andrew J. Friskop ◽  
Thomas J. Gulya ◽  
Robert M. Harveson ◽  
Ryan M. Humann ◽  
Maricelis Acevedo ◽  
...  

Puccinia helianthi, causal agent of sunflower rust, is a macrocyclic and autoecious pathogen. Widespread sexual reproduction of P. helianthi was documented in North Dakota and Nebraska for the first time in 2008 and has since frequently occurred. Concurrently, an increase in sunflower rust incidence, severity, and subsequent yield loss on sunflower has occurred since 2008. Rust can be managed with resistance genes but determination of virulence phenotypes is important for effective gene deployment and hybrid selection. However, the only P. helianthi virulence data available in the United States was generated prior to 2009 and consisted of aggregate virulence phenotypes determined on bulk field collections. The objective of this study was to determine the phenotypic diversity of P. helianthi in the United States. P. helianthi collections were made from cultivated, volunteer, and wild Helianthus spp. at 104 locations across seven U.S. states and one Canadian province in 2011 and 2012. Virulence phenotypes of 238 single-pustule isolates were determined on the internationally accepted differential set. In total, 29 races were identified, with races 300 and 304 occurring most frequently in 2011 and races 304 and 324 occurring most frequently in 2012. Differences in race prevalence occurred between survey years and across geography but were similar among host types. Four isolates virulent to all genes in the differential set (race 777) were identified. The resistance genes found in differential lines HA-R3 (R4b), MC29 (R2 and R10), and HA-R2 (R5) conferred resistance to 96.6, 83.6, and 78.6% of the isolates tested, respectively.


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