scholarly journals First Report of Phakopsora pachyrhizi Adapting to Soybean Genotypes with Rpp1 or Rpp6 Rust Resistance Genes in Field Plots in the United States

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 ◽  
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 ◽  
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 ◽  
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.


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.


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 ◽  
2008 ◽  
Vol 92 (5) ◽  
pp. 767-771 ◽  
Author(s):  
T. L. Slaminko ◽  
M. R. Miles ◽  
R. D. Frederick ◽  
M. R. Bonde ◽  
G. L. Hartman

Phakopsora pachyrhizi, the causal organism of soybean rust, was first found in the continental United States in 2004 and has been found on soybean, kudzu, Florida beggarweed, and three Phaseolus species in the field. The pathogen has been reported to occur on more than 90 legume species worldwide and it is likely to infect native and introduced legume species in the United States. The objective of this study was to determine if 176 species representing 57 genera of legumes, the majority of which are either native or naturalized to soybean-growing areas of the United States, could be hosts of P. pachyrhizi. Between one and three accessions of each species, a total of 264 accessions, were inoculated with a mixture of four isolates of P. pachyrhizi. Severity and sporulation were rated on a 1-to-5 scale at 14 and 28 days after inoculation. P. pachyrhizi was confirmed by the presence of sporulating uredinia and/or immunological assay on 65 new species in 25 genera; 12 of these genera have not been reported previously as hosts. Many of the newly identified hosts grow in the southern United States, and like kudzu, could serve as overwintering hosts for P. pachyrhizi.


Plant Disease ◽  
2009 ◽  
Vol 93 (6) ◽  
pp. 593-598 ◽  
Author(s):  
M. R. Bonde ◽  
S. E. Nester ◽  
W. F. Moore ◽  
T. W. Allen

Soybean rust, caused by Phakopsora pachyrhizi, was first discovered in the continental United States in the fall of 2004. The potential for economic loss in the United States hinges largely on whether or not the pathogen can survive winters in the absence of soybean. Kudzu (Pueraria lobata) is known to be a host for P. pachyrhizi in Asia and South America and is widely distributed in the southern United States. This study examined reactions of kudzu collected from several areas of the southeastern United States to three isolates of P. pachyrhizi, one each from Alabama, Louisiana, and Brazil. Susceptible tan (TAN) lesions, resistant reddish-brown (RB) lesions, and immune (IM) response, previously described on soybean, were produced on kudzu based on the evaluation of 125 plants. However, in contrast to soybean, the RB response on kudzu was common, with approximately 50% frequency. IM responses to at least one isolate were observed on five individual plants, and two plants were immune to all three pathogen isolates used in the test. TAN lesions averaged 3.2 uredinia per lesion with an average diameter per uredinium of 121 μm. In contrast, RB lesions had an average of 0.3 uredinia per lesion with an average uredinial diameter of 77 μm. In 25 of 39 (64%) instances in which multiple plants were tested from a site, each reacted the same to the individual pathogen isolates. This suggested a tendency for plants at specific sites to be genetically identical with respect to rust reaction. Only 19 of 125 (15%) individual plants produced a different reaction to one isolate than to the other two isolates. When four kudzu plants previously shown to produce only TAN lesions to P. pachyrhizi isolates Alabama 04-1, Brazil 01-1, and Louisiana 04-1 were inoculated with eight additional isolates from several areas of the world, all 11 isolates produced only TAN lesions. Likewise, when five other plants previously shown to produce only RB lesions when inoculated with the three isolates were inoculated with the 11 isolates, all produced only RB lesions. These results suggest that susceptibility or resistance to P. pachyrhizi in individual kudzu plants often is broad, extending over a wide range of P. pachyrhizi isolates.


Plant Disease ◽  
2006 ◽  
Vol 90 (3) ◽  
pp. 380-380 ◽  
Author(s):  
C. L. Harmon ◽  
P. F. Harmon ◽  
T. A. Mueller ◽  
J. J. Marois ◽  
G. L. Hartman

Soybean rust caused by Phakopsora pachyrhizi H. Sydow & Sydow was first reported in the continental United States during 2004 (2). By 10 November 2005, the disease was confirmed in eight southern states (Florida, Georgia, Alabama, Mississippi, South Carolina, North Carolina, Louisiana, and Texas). Diagnoses have been based on visual observation of uredinia and urediniospores of the pathogen followed by polymerase chain reaction confirmation. On 10 November 2005, uredinia and telia were identified on leaves of kudzu (Pueraria lobata) in central Florida. Telia first were noted as dark brown-to-black flecks on the abaxial leaf surface intermingled with abundant tan-to-light brown uredinia. Of 200 leaves examined, 143 (72%) had telia. The number of telia ranged from a few (1/cm2) that were scattered to many (73/cm2). Telia were approximately the same diameter as uredinia, but were appressed to the leaf surface and pigmented. Twenty telia were excised from host tissue with the aid of a dissecting microscope and a 20 gauge hypodermic needle. Telia averaged 89 × 100 μm (n = 20, σ = 17 and 16 μm, respectively). Four telia were crushed and five teliospores from each averaged 4.3 × 8.3 μm (n = 20, σ = 0.5 and 0.9 μm, respectively). Pale yellowish brown-to-hyaline teliospores were similar in color to urediniospores. Observations matched descriptions by Ono et al. (1). To our knowledge, this is the first report of the telial stage of P. pachyrhizi in the United States. References: (1) Y. Ono et al. Mycol. Res. 96:825, 1992. (2) R. W. Schneider et al. Plant Dis. 89:774, 2005.


Plant Disease ◽  
2009 ◽  
Vol 93 (8) ◽  
pp. 847-847 ◽  
Author(s):  
Ma. de Jesús Yáñez Morales ◽  
I. Alanis Martínez ◽  
J. Manuel Soto Rocha ◽  
D. K. Malvick ◽  
J. E. Kurle ◽  
...  

Soybean rust caused by Phakopsora pachyrhizi Syd. & P. Syd is a destructive foliar disease of soybean (Glycine max L), which was first confirmed in North America in Louisiana during 2004 (4). Soybean rust (SBR) has also been reported late in the growing season as far north as Illinois, Indiana, and Iowa. SBR was first confirmed in Mexico in 2005 in the state of San Luis Potosi on soybean (3) and subsequently reported in the states of Tamaulipas, Veracruz, and the southwestern coast of Chiapas. Symptoms of SBR were observed on leaves of multiple, nearly mature soybean plants near the city of Campeche (19.72796°N, 90.0771°W) on the Gulf Coast of the Yucatan Peninsula during November 2008. Angular and irregular chlorotic lesions on leaves contained necrotic spots and pale brown, erumpent, cone-like uredinia with a central opening. Ellipsoid to obovoid, echinulate, light tan urediniospores (10 to 13 × 16 to 18 μm) were observed microscopically. DNA was extracted from leaf tissue containing uredinia and from asymptomatic tissue with the DNeasy Plant Mini Kit (Qiagen, Valencia, CA). P. pachyrhizi was confirmed in the symptomatic leaves by a PCR assay with Ppm1/Ppa2 primers, but not from the asymptomatic leaves (1). Subsequently, the DNA extracted from symptomatic and asymptomatic leaf tissues was tested again in another laboratory by a specific quantitative PCR assay (1), and positive results for the presence of soybean rust were obtained only from the symptomatic tissue. As a final confirmatory step, amplified DNA from the PCR assay was sequenced, and the results matched P. pachyrhizi sequences in the GenBank database. To our knowledge, these observations confirm for the first time the presence of P. pachyrhizi in the state of Campeche of southern Mexico. Although it was confirmed on soybean during 2008, it is not known how long the pathogen has been present or which other hosts may be infected there. The presence of SBR on the Yucatan Peninsula is significant because of its potential effects on local plant hosts. In addition, the climate allows possible year-round survival of the pathogen and long-distance transport of urediniospores to the United States. Potential transport of SBR spores from this part of Mexico to the United States has been reported through the application of NOAA's HYSPLIT (Hybrid Single Particle Lagrangian Integrated Transport) model and atmospheric back-trajectory analysis (2). References: (1) R. D. Frederick et al. Phytopathology 92:217, 2002. (2) S. V. Krupa et al. Plant Dis. 90:1254, 2006. (3) A. C. Rodriguez et al. Plant Dis. 90:1260, 2006. (4) R. W. Schneider et al. Plant Dis. 89:774, 2005.


Plant Disease ◽  
2006 ◽  
Vol 90 (7) ◽  
pp. 970-970 ◽  
Author(s):  
T. N. Lynch ◽  
J. J. Marois ◽  
D. L. Wright ◽  
P. F. Harmon ◽  
C. L. Harmon ◽  
...  

Phakopsora pachyrhizi Syd. & P. Syd., the cause of soybean rust, was first observed in the continental United States in November 2004 (2). During the growing season of 2005, P. pachyrhizi was confirmed on soybean (Glycine max) and/or kudzu (Pueraria montana) in nine states in the southern United States. It is known that P. pachyrhizi has a much larger host range within the Fabaceae family. On 29 September 2005 in Quincy, FL, 45 entries of mostly large-seeded legumes were planted next to soybeans that were infected with P. pachyrhizi. Several seeds of each entry were planted on one hill. Soybean plants growing adjacent to these potential hosts had 15 to 25% of the leaf area affected, 95% incidence, and 73% defoliation on 16 November. On 7 December 2005, all the plants of Phaseolus coccineus L. (scarlet runner bean, PI311827), Phaseolus lunatus L. (lima bean, PI583558), and two Phaseolus vulgaris L. (kidney bean) cvs. Red Hawk and California Early Light Red Kidney (CELRK) were found to have leaves with suspected rust lesions. These plants were at physiological maturity but had not senesced. None of the hosts had been inoculated other than from spores produced by the adjacent rust-infected soybean plants or from unknown locations. On the basis of microscopic examination, suspected infected leaves from plants of the Phaseolus spp. had rust pustules characteristic of P. pachyrhizi uredinia. Uredinia were counted within a randomly selected 2-cm2 area of one leaf of each sample. The mean and range of uredinia per lesion for Phaseolus coccineus was 29 uredinia with a range of 0 to 3 uredinia per lesion, Phaseolus lunatus had 2 uredinia with 0 to 1 uredinium per lesion, Phaseolus vulgaris cv. Red Hawk had 22 uredinia with 0 to 5 uredinia per lesion, and Phaseolus vulgaris cv. CELRK had 43 uredinia with 0 to 4 uredinia per lesion. Polymerase chain reactions using two sets of primers (Ppa1/Ppa2 and Pme1/Pme2) were performed on DNA extracted from leaves of the three species with sporulating rust pustules (1). The results of these tests and further tests conducted by the USDA/APHIS confirmed that P. pachyrhizi was the causal organism for the observed rust. References: (1) P. F. Harmon et al. On-line publication. doi:10.1094/PHP-2005-0613-01-RS. Plant Health Progress, 2005. (2) R. W. Schneider et al. Plant Dis. 89:774, 2005.


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