Soybean rust, caused by Phakopsora pachyrhizi Syd., first was observed in the continental United States during 2004 on soybean (Glycine max (L.) Merr.) in Louisiana (4), and on kudzu (Pueraria montana (Lour.) Merr. var. lobata (Willd.) Maesen & Almeida) in Florida (2). Kudzu is a leguminous weed that is prevalent in the southern United States with its range extending northward into other states including Illinois. In October 2009, a kudzu patch located in Pulaski County in southern Illinois was investigated for the presence of soybean rust. Twenty-five leaflets were collected, and the abaxial sides of leaflets were evaluated visually for the presence of uredinia with a dissecting microscope. Uredinia and urediniospores were found on two leaflets. When viewed with a compound microscope, urediniospores were hyaline, echinulate, and measured 20 × 25 μm. On the basis of uredinia and urediniospores, the disease tentatively was identified as soybean rust caused by P. pachyrhizi. To confirm the identification, one leaflet with pustules was assayed with a Soybean Rust QuickStix Diagnostic Kit (Envirologix, Portland, ME). For the other leaflet, the area of the pustule was excised (approximately 28 mm2) and an area of the leaflet at the margin on the opposite half of the leaflet with no visible pustule (approximately 54 mm2) was excised. DNA was extracted from the excised areas of the leaflet for confirmation by quantitative PCR (Q-PCR) using primers and probe specific to P. pachyrhizi and P. meibomiae (Arthur) Arthur (1). Both the QuickStix Diagnostic Kit and the Q-PCR confirmed the diagnosis as soybean rust caused by P. pachyrhizi. Q-PCR also suggested the presence of a nonsporulating latent rust infection on the same kudzu leaflet at the margin on the opposite side of the midrib. Soybean rust first was confirmed on soybean in Illinois in 2006 (3), but to our knowledge, this is the first observation of the disease on kudzu in the state. This report confirms that at least some kudzu plants in Illinois are susceptible to soybean rust and that latent kudzu infection may exist without outward signs of the fungus. Currently, this is the most northern observation of soybean rust on kudzu in North America. It is unknown what role, if any, Illinois kudzu will play in the epidemiology of soybean rust in the state. Since kudzu tops die after the first frost, there is no expectation of P. pachyrhizi to overwinter in Illinois on kudzu as it does in some states adjacent to the Gulf of Mexico.

Appressoria of some plant pathogenic fungi accumulate turgor pressure that produces a mechanical force enabling the direct penetration of hyphae through the epidermis. Melanin has been reported to function as an impermeable barrier to osmolytes, which allow appressoria to accumulate high turgor pressure. Deficiency of melanin in appressoria has been shown to reduce turgor pressure and compromise the infection process. In Phakopsora pachyrhizi, the soybean rust pathogen, the appressoria are transparent. Our objective was to determine if melanin inhibitors would alter appressorial turgor pressure and if a melanin layer would form specifically between the appressorial cell wall and plasma membrane. We used two melanin biosynthesis inhibitors and found that these melanin inhibitors did not reduce turgor pressure or compromise the infection process. In addition, the turgor pressure of P. pachyrhizi appressoria ranged from 5 to 6 MPa based on extracellular osmolytes used to simulate different osmotic pressures. Transmission electron microscopy also showed the absence of a melanin layer between the appressorial cell wall and plasma membrane. This is the first report showing that turgor pressure accumulation of P. pachyrhizi appressoria was independent of melanin.

Soybean rust (SBR), caused by Phakopsora pachyrhizi Sydow, is one of the most economically important and destructive diseases of soybean [Glycine max (L.) Merr.] and the discovery of novel SBR resistance genes is needed because of virulence diversity in the pathogen. The objectives of this research were to map SBR resistance in plant introduction (PI) 561356 and to identify single nucleotide polymorphism (SNP) haplotypes within the region on soybean chromosome 18 where the SBR resistance gene Rpp1 maps. One-hundred F(2:3) lines derived from a cross between PI 561356 and the susceptible experimental line LD02-4485 were genotyped with genetic markers and phenotyped for resistance to P. pachyrhizi isolate ZM01-1. The segregation ratio of reddish brown versus tan lesion type in the population supported that resistance was controlled by a single dominant gene. The gene was mapped to a 1-cM region on soybean chromosome 18 corresponding to the same interval as Rpp1. A haplotype analysis of diverse germplasm across a 213-kb interval that included Rpp1 revealed 21 distinct haplotypes of which 4 were present among 5 SBR resistance sources that have a resistance gene in the Rpp1 region. Four major North American soybean ancestors belong to the same SNP haplotype as PI 561356 and seven belong to the same haplotype as PI 594538A, the Rpp1-b source. There were no North American soybean ancestors belonging to the SNP haplotypes found in PI 200492, the source of Rpp1, or PI 587886 and PI 587880A, additional sources with SBR resistance mapping to the Rpp1 region.

Soybean rust, caused by Phakopsora pachyrhizi, occurs concomitantly wherever soybean is grown in the tropical and subtropical regions of the world. After reports of its first occurrence in Brazil in 2001 and the continental United States in 2004, research on the disease and its pathogen has greatly increased. One area of research has focused on capturing urediniospores, primarily by rain collection or wind traps, and detecting them either by microscopic observations or by immunological or molecular techniques. This system of detection has been touted for use as a potential warning system to recommend early applications of fungicides. One shortcoming of the method has been an inability to determine whether the spores are viable. Our study developed a method to detect viable P. pachyrhizi urediniospores using an immunofluorescence assay combined with propidium iodide (PI) staining. Antibodies reacted to P. pachyrhizi and other Phakopsora spp. but did not react with other common soybean pathogens or most other rust fungi tested, based on an indirect immunofluorescence assay using fluorescein isothiocyanate-labeled secondary antibodies. Two vital staining techniques were used to assess viability of P. pachyrhizi urediniospores: one combined carboxy fluorescein diacetate (CFDA) and PI, and the other utilized (2-chloro-4-[2,3-dihydro-3-methyl-(benzo-1,3-thiazol-2-yl)-methylidene]-1-phenylquinolinium iodide] (FUN 1). Using the CFDA-PI method, viable spores stained green with CFDA and nonviable spores counterstained red with PI. Using the FUN 1 method, cylindrical intravacuolar structures were induced to form within metabolically active urediniospores, causing them to fluoresce bright red to reddishorange, whereas dead spores, with no metabolic activity, had an extremely diffused, faint fluorescence. An immunofluorescence technique in combination with PI counterstaining was developed to specifically detect viable P. pachyrhizi urediniospores. The method is rapid and reliable, with a potential for application in forecasting soybean rust based on the detection of viable urediniospores.

Soybean rust, caused by the fungus Phakopsora pachyrhizi, is an economically important disease of soybean with potential to cause severe epidemics resulting in significant yield losses. Host resistance is one of the management tools to control this disease. This study compared soybean genotypes exhibiting immunity, complete and incomplete resistance, and susceptibility to an isolate of P. pachyrhizi based on visual assessment of reaction type, other visual traits such as sporulation, quantitative measurements of the amount of fungal DNA (FDNA) present in leaf tissues, and data on infection and colonization levels. Soybean genotype UG5 (immune), and plant introduction (PI) 567102B and PI 567104B (complete resistance) had lower quantities of uredinia and FDNA than four other genotypes with incomplete resistance. Based on microscopic observations, early events of spore germination, appressorium formation, and fungal penetration of the epidermis occurred within 24 h postinoculation and were similar among the tested soybean genotypes. Differences in infection among the genotypes were evident once the hyphae penetrated into the intercellular spaces between the mesophyll cells. At 2 days after inoculation (dai), soybean genotype Williams 82 had a significantly (P < 0.05) higher percentage of hyphae in the mesophyll tissue than other soybean genotypes, with UG5 having significantly (P < 0.05) lower percentages than all of the other soybean genotypes at 3, 4, and 5 dai. The percentage of interaction sites with mesophyll cell death was significantly (P < 0.05) higher in UG5 than other genotypes at 3, 4, and 5 dai. There was a significant positive correlation (r = 0.30, P < 0.001) between quantities of hyphae in the mesophyll cells and FDNA. These results demonstrated that incompatible soybean–P. pachyrhizi interaction results in restricted hyphal development in mesophyll cell tissue, likely due to hypersensitive apoptosis.

Phakopsora pachyrhizi Sydow, the causal fungus of soybean (Glycine max (L.) Merrill) rust, occurs in most soybean-growing areas of the world except continental North America. Initial studies on soybean rust isolates from the Western Hemisphere indicated that they were different than isolates from the Eastern Hemisphere. In 1992, the Eastern Hemisphere species, P. pachyrhizi, and the Western Hemisphere species, P. meibomiae, were established for the soybean rust fungi based on morphological differences. The first molecular differentiation of the two species was reported in 2002. A number of studies have reported the occurrence of race in P. pachyrhizi either on soybeans or on other hosts. In 1984, a set of four native Australian Glycine species were used to identify six different virulence combinations of P. pachyrhizi. Much of the research on differentiating isolates on soybean was completed in a containment facility at in the U.S. Genetic characterization on four plant introductions (PIs) indicated the occurrence of four independently inherited dominant genes. These genes are known to be effective to a limited number of isolates. There are many studies that need to be completed to determine if all isolates respond equally in terms of survival, urediniospore production, telia formation, and host range under different environments. Over the next few years, our understanding of pathogen diversity will increase as more concerted research efforts take place in different parts of the world.

Quantitative polymerase chain reaction (Q-PCR) utilizing specific primer sequences and a fluorogenic, 5′-exonuclease linear hydrolysis probe is well established as a detection and identification method for Phakopsora pachyrhizi and P. meibomiae, two rust pathogens of soybean. Because of the extreme sensitivity of Q-PCR, the DNA of single urediniospores of these fungi can be detected from total DNA extracts of environmental samples. However, some DNA preparations unpredictably contain PCR inhibitors that increase the frequency of false negatives indistinguishable from true negatives. Three synthetic DNA molecules of arbitrary sequence were constructed as multiplexed internal controls (ICs) to cull false-negative results by producing a positive signal to validate the PCR process within each individual reaction. The first two, PpaIC and PmeIC, are a single-stranded oligonucleotide flanked by sequences complementary to the primers of either the P. pachyrhizi or P. meibomiae primary assay but hybridizing to a unique fluorogenic probe; the third contains unique primer- and probe-binding sequences, and was prepared as a cloned DNA fragment in a linearized plasmid. These ICs neither qualitatively nor quantitatively affected their primary assays. PpaIC and PmeIC were shown to successfully identify false-negative reactions resulting from endogenous or exogenous inhibitors, and can be readily adapted to function in a variety of diagnostic Q-PCR assays; the plasmid was found to successfully validate true negatives in similar Q-PCR assays for other soybean pathogens, as well as to function as a tracer molecule during DNA extraction and recovery.

Soybean rust, caused by Phakopsora pachyrhizi, may be the most important foliar disease of soybean. Within the last 10 years, the fungus has moved to many new geographical locations via spread of airborne urediniospores. The objective of this study was to determine the relationship between urediniospore viability and exposure to solar radiation. Urediniospores of P. pachyrhizi were exposed in Capitán Miranda, Paraguay, to determine the deleterious effects of sunlight. Concomitant total solar (0.285 to 2.8 µm) and ultraviolet (0.295 to 0.385 µm) irradiance measurements were used to predict urediniospore germination. Urediniospores exposed to doses of solar and ultraviolet (UV) radiation ≥27.3 MJ/m2 and ≥1.2 MJ/m2, respectively, did not germinate. The proportions of urediniospores that germinated, normalized with respect to the germination proportion for unexposed urediniospores from the same collections, were a linear function of solar irradiance (R2 = 0.83). UV measurements predicted normalized germination proportions equally well. Results of inoculation experiments with exposed P. pachyrhizi urediniospores supported the results of the germination trials, although the effects of moderate levels of irradiance varied. The relationship between urediniospore viability and exposure to solar radiation has been incorporated into the U.S. Department of Agriculture’s soybean rust aerobiological model that provides North American soybean growers with decision support for managing soybean rust.

Soybean rust, caused by Phakopsora pachyrhizi, is one of the most important foliar diseases of soybean worldwide. In a series of experiments, multiple objectives were addressed to determine the (i) longevity of detached soybean leaves, (ii) reproductive capacity of uredinia on leaves inoculated and/or incubated on the abaxial versus adaxial side of the leaf, (iii) reproductive capacity of uredinia and urediniospore germination when spores were harvested at regular intervals or all at once, and (iv) effect of temperature and relative humidity (RH) on urediniospore germination. A detached-leaf assay using agar medium amended with 6-benzylaminopurine performed better in retarding leaf chlorosis than filter paper alone among five soybean genotypes. Among the three susceptible genotypes tested, detached leaves of cv. Williams 82 had the lowest level of leaf chlorosis and often allowed for the greatest urediniospore production and germination rate. Temperature and RH played significant roles in survival of urediniospore as measured by germination rates. Viable urediniospores were harvested from infected soybean leaves maintained at room temperature (23 to 24°C at 55 to 60% RH) for up to 18 days, whereas freshly harvested urediniospores that were desiccated for 12 h before being placed in vials and maintained at room temperature remained viable for up to 30 days. Urediniospore hydration was the major factor for the dormancy reversion; thermal shock with hydration and no thermal shock with hydration treatments had consistently similar urediniospore germination rates. In the RH experiment, urediniospores harvested from inoculated leaf pieces maintained at 85% RH had the highest germination rates compared with higher and lower RH. Improvement in P. pachyrhizi cultural techniques and understanding of urediniospore survival will enhance our knowledge of the pathogen biology, host-plant relationship, and conditions that favor the infection, reproduction, and survival of the pathogen.

The introduction of Phakopsora pachyrhizi, the cause of soybean rust, into the U.S. is a classic case of a pathogen introduction that became established in a new geographical region on a perennial overwintering host (kudzu). The objective of our study was to classify the pathogenic variation of P. pachyrhizi isolates collected in the U.S., and to determine the spatial and temporal associations therof. A total of 72 isolates of P. pachyrhizi collected from infected kudzu and soybean leaves in the U.S. were purified, and 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, from collections made in 2008, and from both hosts (kudzu or soybean). Shannon’s index based on the number of pathotypes, indicated that isolates from the South region were more diverse (H values of 0.83) compared to the isolates collected in other regions. This study establishes a baseline of pathogenic variation of P. pachyrhizi in the U.S. that can be further compared to 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.

Soybean rust, caused by Phakopsora pachyrhizi, is an important disease in Nigeria and many other soybean-producing countries world-wide. To determine the geographical distribution of soybean rust in Nigeria, soybean fields were surveyed in the Derived Savanna (DS), Northern Guinea Savanna (NGS), and Southern Guinea Savanna (SGS) agroecological zones in Nigeria between 2004 and 2006. Disease severity in each zone was determined and analyzed using geostatistics. Prevalence of infected fields and disease severity in surveyed fields were significantly (P < 0.05) different between geographical zones with both variables being higher in the DS zone than in either NGS or SGS zones. Geostatistical analysis indicated that the spatial influence of disease severity at one location on severity at other locations was between 75 and 120 km. An exponential model best described the relationship between semivariance and lag distance when rust severity was high. Spatial interpolation of rust severity showed that locations in the DS zone were more conducive for the rust epidemic compared to areas in the NGS zone. In the 2005 survey, 116 purified isolates were established in culture on detached soybean leaves. To establish the nature of pathogenic variation in P. pachyrhizi, a set of four soybean accessions with Rpp(1), Rpp(2), Rpp(3), and Rpp(4) resistance genes, two highly resistant and two highly susceptible genotypes were inoculated with single uredinial isolates. Principal component analysis on the number of uredinia per square centimeter of leaf tissue for 116 isolates indicated that an adequate summary of pathogenic variation was obtained using only four genotypes. Of these four, PI 459025B (with Rpp(4) gene) and TG× 1485-1D had the lowest and highest number of uredinia per square centimeter, respectively. Based on cluster analysis of the number of uredinia per square centimeter, seven pathotype clusters were determined. Isolates in cluster III were the most virulent, while those in cluster IV were the least virulent. Shannon’s index (H) revealed a more diverse pathogen population in the DS zone (H = 1.21) compared to the rust population in SGS and NGS with H values of 1.08 and 0.91, respectively. This work will be useful in breeding and management of soybean rust by facilitating identification of resistant genotypes and targeting cultivars with specific resistance to match prevailing P. pachyrhizi pathotypes in a given geographical zone.

An important mechanism for genetic diversity in filamentous fungi is hyphal anastomosis and the formation of heterokaryons. In this study, we observed fusion of germ tubes in germinating urediniospores of Phakopsora pachyrhizi resulting in a complex hyphal network. Staining of the germ tubes derived from P. pachyrhizi urediniospores with 4’, 6-diamidino-2-phenylindole (DAPI) showed migration of nuclei through the network resulting in multinucleate hyphae. Short bridges connecting the hyphal network tubes were also observed. Our study provides the first evidence of germ tube and hyphal anastomosis, and nuclear migration in P. pachyrhizi. Considering the lack of a known sexual stage of P. pachyrhizi, this hyphal anastomosis followed by the parasexual cycle may explain the genetic diversity in virulence among populations of P. pachyrhizi.

Asian soybean rust (ASR), caused by Phakopsora pachyrhizi and recently discovered for the first time in continental United States, has been of concern to the U.S. agricultural industry for more than 30 years. Since little soybean rust resistance is known, and resistance is often difficult to detect or quantitate, we initiated a project to develop a better, more quantitative, method. The methodology determined the average numbers and diameters of uredinia in lesions that developed on leaves of inoculated plants 14 days after inoculation. It was used to compare virulence of P. pachyrhizi isolates from Asia and Australia and P. meibomiae from Puerto Rico and Brazil, collected as many as 30 years earlier, with isolates of P. pachyrhizi recently collected from Africa or South America. Susceptible reactions to P. pachyrhizi resulted in tan-colored lesions containing 1 to 14 uredinia varying greatly in size within individual lesions. In contrast, on these same genotypes at the same time of year, resistance to other P. pachyrhizi isolates was typified by 0 to 6 small uredinia in reddish-brown to dark-brown lesions. Using appropriate rust resistant and rust susceptible genotypes as standards, examination of uredinia 14 days after inoculation allowed quantitative comparisons of sporulation capacities, one measure of susceptibility or resistance to soybean rust. The study verified the presence and ability to detect all known major genes for resistance to soybean rust in the original sources of resistance. It demonstrated that soybean lines derived from the original PI sources, and presumed to possess the resistance genes, in actuality may lack the gene or express an intermediate reaction to the rust pathogen. We suggest that a determination of numbers and sizes of uredinia will detect both major gene and partial resistance to soybean rust.

Soybean rust, caused by Phakopsora pachyrhizi (H. Sydow & Sydow), is a destructive soybean [Glycine max (L.) Merr.] disease and identification of new resistance genes is essential for effective rust management. Our research objectives were to map and confirm the location of resistance gene(s) in PI 594538A using a population of 98 F_3:4 lines from a cross between PI 594538A (reddish-brown [RB] lesions) and the susceptible cultivar Loda (tan [TAN] lesions). The lines were inoculated with the P. pachyrhizi isolate ZM01-1 from Zimbabwe. The RB resistance in PI 594538A mapped on linkage group G as a single dominant gene. This gene is likely an allele of Rpp1 or a new closely linked gene because it mapped within 1 cM of Rpp1 and ZM01-1 produced RB lesions on PI 594538A and TAN lesions on PI 200492, the original source of Rpp1. The mapping of the new Rpp1 allele, named Rpp1-b, was confirmed in a second population.

Accessions of 12 perennial Glycine species were evaluated for resistance to Phakopsorapachyrhizi, the causal agent of soybean rust. A total of 23% of the accessions were resistant, 18% were moderately resistant, and 58% were susceptible. In two experiments, 59 and 40% of the accessions of G. tabacina (2/n=80) were resistant. Resistance to P. pachyrhizi was identified in accessions of G. argyrea, G. canescens, G. clandestina, G. latifolia, G. microphylla, and G. tomentella, but not in accessions of G. arenaria, G. cyrtoloba, G. curvata, and G. falcata.

Phakopsora pachyrhizi was inoculated on two soybean (Glycine max) genotypes at three different reproductive growth stages (GS) in four trials. Leaf rust was more severe on Taita Kaohsiung No. 5 (TK 5), a commercial cultivar, than on SRE-B15-A (B15 A), a genotype selected for tolerance to leaf rust. At GS R6, the percentage of leaf area infected ranged from 14 to 95% for TK 5 and from 0 to 34% for B15 A. Values for area under disease progress curve (AUDPC) were significantly greater for TK 5 than B15 A. Yields in fungicide-protected plots ranged from 2,312 to 3,546 kg/ ha and were not significantly different between the genotypes. Average yields of plants inoculated at GS Rl were reduced by 62 and 22% and seed weights by 35 and 14% for TK 5 and B15 A, respectively, compared with fungicide-protected plots. Regressions of yield percentage of fungicide-protected plants on disease severity assessments at GS R6, AUDPC, and area under the green leaf area curve were significant for both genotypes.

Soybean rust (SBR), caused by Phakopsora pachyrhizi, was first discovered in North America in 2004 and has the potential to become a major soybean [Glycine max (L.) Merr.] disease in the USA. Currently, four SBR resistance genes have been identified but not mapped on the soybean genetic linkage map. One of these resistance genes is the Rpp1 gene, which is present in the soybean accession PI 200492. The availability of molecular markers associated with Rpp1 will permit marker-assisted selection and expedite the incorporation of this gene into U.S. cultivars. We compared simple sequence repeat (SSR) markers between ‘Williams 82’ and the BC5 Williams 82 isoline L85-2378, which contains the Rpp1 resistance allele from the soybean accession PI 200492, for candidate regions that might contain Rpp1. One candidate region was found with the SSR marker BARC_Sct_187 on linkage group G. A population of BC6F_2:3 lines segregating for the Rpp1 resistance locus was genotyped in this region on linkage group G followed by inoculation with the P. pachyrhizi isolate India 73-1 in the USDA-ARS Biosafety Level 3 Plant Pathogen Containment Facility at Ft. Detrick, MD. The Rpp1 gene was mapped between SSR markers BARC_Sct_187 and BARC_Sat_064 on linkage group G.

Five hundred thirty soybean accessions from maturity groups (MG) III through IX were evaluated for resistance to Phakopsora pachyrhizi in a replicated field trial at Centro Regional de Investigación Agrícola in Capitán Miranda, Itapúa, Paraguay during the 2005–06 season. Soybean rust severities of individual accessions ranged from 0% (resistant) to 30.0% (susceptible). In MG III and IV, the most resistant accessions were PI 506863, PI 567341, and PI 567351B, with severities less than 1.2%. In MG V, the most resistant accessions were PI 181456, PI 398288, PI 404134B, and PI 507305, with severities less than 0.3%. In MG VI, the most resistant accessions were PI 587886, PI 587880A, and PI 587880B, with severities less than 0.3%. In MG VII and VIII, the most resistant were PI 587905 and PI 605779E, with severities less than 1.0%. In MG IX, the most resistant accessions were PI 594754, PI 605833, PI 576102B, and PI 567104B, with severities less than 1.0%. The resistance in 10 selected accessions from MG VI, VII, VIII, and XI was confirmed in subsequent greenhouse and field experiments where severities of 0.4% or less and reddish-brown lesions with sporulation levels less than 3.0 were observed. These accessions, with low severities in the adult plant field evaluation, may be new sources of resistance to P. pachyrhizi.

Asian soybean rust, Phakopsora pachyrhizi, has been an important pathogen of soybean in Asia with yield losses of 40 to 80% commonly reported. The pathogen has moved into Africa, where it was reported in Uganda in 1996, then Zimbabwe (1998) and South Africa (2001). The pathogen was first found South America in Paraguay and then Brazil during the 2001 growing season. A set of 174 soybean accessions was evaluated against local soybean rust populations in field or greenhouse studies in Brazil, China, Paraguay, and Thailand. The materials were also evaluated in the USDA BSL-3 containment greenhouse in Ft. Detrick, MD against a mixed collection of P. pachyrhizi from Brazil, Paraguay, Thailand and Zimbabwe. Among the set were soybeans that had previously been reported to have resistance to either P. pachyrhizi or P. meibomia, including the sources of the four identified resistance genes. The pathogen is known to have a complex and diverse virulence pattern with many phenotypes seen within a field collection. This was observed as mixed resistant (RB) and susceptible reactions on several lines within each location. Disease severity and reaction phenotypes on individual lines differed by location. These differences were due to local environmental conditions, which reduced rust severity as well as differences in the virulence of the rust population at each location. No lines were found to be resistant at all locations.

Soybean rust (Phakopsora pachyrhizi) has been reported on common bean (Phaseolus vulgaris) in Asia, South Africa, and the United States. However, there is little information on the interaction of individual isolates of Phakopsora pachyrhizi with common bean germplasm. A set of 16 common bean cultivars with known genes for resistance to Uromyces appendiculatus, the causal agent of common bean rust, three soybean accessions that were sources of the single gene resistance to P. pachyrhizi, and the moderately susceptible soybean ‘Ina’ were evaluated using seedlings inoculated with six isolates of P. pachyrhizi. Among the common bean cultivars, Aurora, Compuesto Negro Chimaltenango, and Pinto 114, were the most resistant to all six P. pachyrhizi isolates, with lower severity, less sporulation, and consistent reddish-brown (RB) lesions associated with resistance in soybean. A differential response was observed among the common bean cultivars, with a cultivar-isolate interaction for both severity and sporulation levels, as well as the presence or absence of the RB lesion type. This differential response was independent of the known genes that condition resistance to U. appendiculatus, suggesting that resistance to P. pachyrhizi was independent of resistance to U. appendiculatus.

The accessions in the USDA Germplasm Collection at the Univ. of Illinois were evaluated to identify soybean germplasm with resistance to soybean rust. The inoculation program used a mixture of four P. pachyrhizi isolates. Of 16,595 accessions rated for rust severity, 3,215 were selected for a second round of evaluation. Of these, 805 were selected for further evaluation. Some of these selected accessions have the potential to provide soybean rust resistance genes that may be useful for incorporation into commercial soybean cultivars.

Soybean resistance to Phakopsora pachyrhizi, the cause of soybean rust, has been characterized by the following three infection types: (i) immune response (IM; complete resistance) with no visible lesions, (ii) resistant reaction with reddish brown (RB) lesions (incomplete resistance), and (iii) susceptible reaction with tan-colored (TAN) lesions. Based on visual assessments of these phenotypes, single gene resistance in soybean to P. pachyrhizi has been documented, but colonization within infected tissues based on fungal DNA (FDNA) levels in different soybean genotypes had not been analyzed. The research used a quantitative polymerase chain reaction (Q-PCR) assay to compare visual disease assessment to FDNA in controlled inoculation experiments using two isolates of P. pachyrhizi. The objective of the first experiment was to compare data from digital visual disease assessment to FDNA from Q-PCR assays using digital visual disease assessment using five resistant soybean genotypes (one IM and four RB) and five susceptible genotypes (TAN). The objective of the second experiment was to quantify FDNA using Q-PCR at different time points after inoculation to determine if levels of fungal colonization differed in five soybean genotypes with different levels of resistance (one IM, two RB, and two TAN). For experiment 1, the numbers of uredinia and uredinia per lesion on four of the five resistant soybean genotypes were lower (P < 0.05) than the other six genotypes. Significant differences (P < 0.05) in FDNA concentrations were found among soybean genotypes with TAN lesions and among soybean genotypes with RB lesions. Soybean cultivar UG5 (IM phenotype) had significantly less (P < 0.05) FDNA than all of the other genotypes. Some genotypes that produced TAN lesions had significantly lower (P < 0.05) or non-significantly different FDNA concentrations compared to those genotypes that produced RB lesions. For experiment 2, the regression of FDNA on days after inoculation was significant (P < 0.01) with positive slopes for all genotypes except for UG5, in which FDNA declined over time, indicating a reduction of fungal colonization. The results of this Q-PCR FDNA screening technique demonstrates its use to distinguish different types of resistance, and could be used to facilitate the evaluation of soybean breeding populations, where precise quantification of incomplete and/or partial resistance is needed to identify and map quantitative trait loci.

Soybean rust, caused by the fungus Phakopsora pachyrhizi, was detected in the continental United States in 2004. Several new sources of resistance to P. pachyrhizi have been identified in soybean (Glycine max); however, there is limited information about their resistance when challenged with additional U.S. and international isolates. Resistance of 20 soybean (G. max) entries was compared after inoculation with 10 P. pachyrhizi isolates, representing different geographic and temporal origins. Soybean entries included 2 universal susceptible cultivars, 4 sources of soybean rust resistance genes (Rpp1–4), and 4 and 10 resistant entries selected from field trials in Paraguay and Vietnam, respectively. Of the known Rpp1–4 sources of resistance, plant introduction (PI) 459025B (Rpp4) produced reddish-brown (RB) lesions in response to all of the P. pachyrhizi isolates, while PI 230970 (Rpp2) produced RB lesions to all isolates except one from Taiwan, in response to which it produced a susceptible tan (TAN) lesion. PI 200492 (Rpp1) and PI 462312 (Rpp3) produced TAN lesions in response to most P. pachyrhizi isolates. The resistant entries selected from Paraguay and Vietnam varied considerably in their responses to the 10 P. pachyrhizi isolates, with M 103 the most susceptible and GC 84058-18-4 the most resistant. The reaction patterns on these resistant entries to the P. pachyrhizi isolates were different compared with the four soybean accessions with the Rpp genes, indicating that they contain novel sources of rust resistance. Among the P. pachyrhizi isolates, TW 72-1 from Taiwan and IN 73-1 from India produced the most susceptible and resistant reactions, respectively, on the soybean entries.

Soybean rust, caused by Phakopsora pachyrhizi, is one of the most important constraints to soybean production worldwide. The absence of high levels of host resistance to the pathogen has necessitated the continued search and identification of sources of resistance. In one set of experiments, 178 soybean breeding lines from the International Institute of Tropical Agriculture were rated for rust severity in the field in 2002 and 2003 at Ile-Ife, Yandev, and Ibadan, Nigeria. Thirty-six lines with disease severity ≤3 (based on a 0-to-5 scale) were selected for a second round of evaluation in 2004 at Ibadan. In the third round of evaluation under inoculated field conditions, 11 breeding lines with disease severity ≤2 were further evaluated for rust resistance at Ibadan in 2005 and 2006. The breeding lines TGx 1835-10E, TGx 1895-50F, and TGx 1903-3F consistently had the lowest level of disease severity across years and locations. In another set of experiments, 101 accessions from the United States Department of Agriculture–Agricultural Research Service and National Agriculture Research Organization (Uganda) were evaluated in the first round in 2005 under inoculated conditions in the screenhouse; 12 accessions with disease severity ≤20% leaf area infected were selected for evaluation in the second round in 2005 and 2006 under inoculated field conditions at Ibadan. Highly significant differences (P < 0.0001 in disease severity were observed among the 101 accessions during this first round of rust evaluation. Significant (P < 0.0001) differences in rust severity and sporulation also were observed among the 12 selected accessions. Accessions PI 594538A, PI 417089A, and UG-5 had significantly (P < 0.05) lower disease severity than all other selected accessions in both years of evaluation, with rust severities ranging from 0.1 to 2.4%. These results indicate that some of the breeding lines (TGx 1835-10E, TGx 1895-50F, and TGx 1903-3F) and accessions (PI 594538A, PI 417089A, and UG-5) would be useful sources of soybean rust resistance genes for incorporation into high-yielding and adapted cultivars.

Fourteen soybean accessions and breeding lines were evaluated for resistance to soybean rust caused by the fungus Phakopsora pachyrhizi. Evaluations were conducted in replicated experiments in growth chambers using detached leaves and under greenhouse and field conditions. In growth-chamber experiments, inoculation of detached leaves with 1 × 106 spores/ml resulted in a significantly (P < 0.0001) higher total number of pustules and spores per unit leaf area than inoculations with lower spore concentrations. Amending agar medium with plant hormones significantly (P < 0.0001) aided retention of green leaf color in detached leaves. Leaf pieces on a medium containing kinetin at 10 mg/liter had 5% chlorosis at 18 days after plating compared with leaf pieces on media amended with all other plant hormones, which had higher levels of chlorosis. Leaf age significantly affected number of pustules (P = 0.0146) and number of spores per pustule (P = 0.0088), and 3- to 4-week-old leaves had a higher number of pustules and number of spores per pustule compared with leaves that were either 1 to 2 or 5 to 6 weeks old. In detached-leaf and greenhouse screening, plants were evaluated for days to lesion appearance, days to pustule formation, days to pustule eruption, lesion number, lesion diameter, lesion type, number of pustules, and spores per pustule in 1-cm2 leaf area. Plants also were evaluated for diseased leaf area (in greenhouse and field screening) and sporulation (in field screening) at growth stage R6. There were significant (P < 0.0001) differences among genotypes in their response to P. pachyrhizi infection in the detached-leaf, greenhouse, and field evaluations. Accessions PI 594538A, PI 417089A, and UG-5 had very low levels of disease compared with the susceptible checks and all other genotypes. Detached-leaf, greenhouse, and field results were comparable, and there were significant correlations between detached-leaf and greenhouse (absolute r = 0.79; P < 0.0001) and between detached-leaf and field resistance (absolute r = 0.83; P< 0.0001) across genotypes. The overall results show the utility of detached-leaf assay for screening soybean for rust resistance.

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-cm(^2) 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. 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.

Asian soybean rust is one of the most devastating diseases of soybean with yield losses of 10 to 100% reported. The disease is found primarily in the lower canopy before flowering and in the middle and upper canopy after flowering. Heavily infected plants often prematurely defoliate causing significant yield losses. Until useful genetic resistance can be identified and moved into commercial cultivars, fungicides will be the primary means to control the disease. There is not much information on fungicide application in soybean. Fungicide use in soybean has been limited to seed treatments and a single late season foliar. With soybean rust the canopy needs to be protected from onset of flowering through pod fill. The research presented is a preliminary summary of the measurement of canopy penetration using high and low water volumes with two fungicides, Bravo and Quadris. Fungicides were applied aerially at 5 and 10 gal/ac in six locations in the southern US and by ground in three locations in the midwest US. In the ground application experiment, air induction, flat fan pointed down, flat fans on drops set to spray 105° and twin jet on forward facing right angle drops set to spray 80° nozzles were compared. Field design was set up as a strip plot with at least three replications per location. Three water sensitive paper strips were placed at mid canopy across the spray swath in three locations the length of the plot. Increased water volume in both ground and aerial application improved fungicide coverage when compared to the lower application volume. Among the nozzle tips evaluated in ground applications, overhead flat fans provided the least fungicide coverage in mid canopy.

This paper focuses on the Asian soybean rust (Phakopsora pachyrhizi), the fungicides so far evaluated to control this disease, including the corresponding country where each fungicide was tested, as well as the individual summaries of application trials and recommendations. The importance of the timing and number of applications to control the disease, and the factors determining the number of applications (length of the reproductive phase of the crop, persistence of the compound and severity of the epidemic) are also emphasized. Among the most effective fungicides from trial done in Southern Africa were flusilazole + carbendazim, difenoconazole, and triadimenol. Trials in South America have identified several triazoles (tebuconazole and tetraconazole) as well as several strobularins and strobularin mixes ( azoxystrobin, pyraclostrobin, pyraclostrobin + boscalid and trifloxystrobin + propoconazole). The recommendations for control of the disease based on the experience in Southern Africa were two applications with the first application times 50 days after planting, at or before flowering, and a second application 20 days later.

The efficacy of fungicides in managing soybean rust was evaluated in 12 environments in South America and southern Africa over three growing seasons from 2002 to 2005. There were differences in final soybean rust severity, defoliation, and yield among the treatments at most locations. In locations where soybean rust was not severe, all the fungicides evaluated reduced severity. In locations where soybean rust was severe, applications of triazole and triazole + strobilurin fungicides resulted in lower severity and higher yields compared with other fungicides. The strobilurin fungicides provided the highest yields in many locations; however, severity tended to be higher than that of the triazole fungicides. There also were differences in yield and severity between the trials with two and three applications of several fungicides, with three applications resulting in less severe soybean rust and higher yields. However, the third application of tebuconazole, tetraconazole, and the mixtures containing azoxystrobin and pyraclostrobin was not needed to maintain yield. These fungicides were among the most effective for managing soybean rust and maintaining yield over most locations.

Soybean rust, caused by Phakopsora pachyrhizi, is a devastating foliar disease of soybean that may cause significant yield losses if not managed by well-timed fungicide applications. To determine the effect of fungicide timing on soybean rust severity and soybean yield, field trials were completed in Paraguay (four locations), the United States (two locations), and Zimbabwe (one location) from 2005 to 2006. Treatments at each location included applications of tebuconazole, pyraclostrobin, or a combination of azoxystrobin + propiconazole, and in some locations pyraclostrobin + tebuconazole at the following soybean growth stages (GS): (i) GS R1 (beginning flowering), (ii) GS R3 (beginning pod), (iii) GS R5 (beginning seed), (iv) GS R1 + R3, (v) GS R3 + R5, and (vi) GS R1 + R3 + R5. Soybean yields from plots treated with fungicides were 16 to 114% greater than yields from no fungicide control plots in four locations in Paraguay, 12 to 55% greater in two locations in the United States, and 31% greater in Zimbabwe. In all locations, rust severity measured over time as area under the disease progress curve (AUDPC) was negatively correlated (r = –0.3, P < 0.0001) to yield. The effectiveness of any given treatment (timing of application and product applied) was often dependent on when rust was first detected and the intensity of its development. For example, when soybean rust was first observed before GS R3 (two locations in Paraguay), the plants in plots treated with a fungicide at GS R1 had the lowest AUPDC values and highest yields. When soybean rust was first observed after GS R3, plants treated with a fungicide at GS R3 and/or GS R5 had the lowest AUDPC values and highest yields with a few exceptions.

Soybean rust (SBR), caused by Phakopsora pachyrhizi, has become established in Africa since the first report in Uganda in 1996 (2). The urediniospores, as windborne propagules, have infested new regions of Africa, initiating SBR in many countries, including Ghana and Democratic Republic of the Congo in 2007 (4) and Tanzania in 2014 (3). No refereed reports have been published about rust in Malawi, but some people have indicated that soybean rust may have been observed as early as 2008. Typical symptoms and signs of SBR, including leaf yellowing and tan, sporulating uredinia, were observed on soybean in May 2014 during field surveys in the major soybean-growing areas of Malawi, including the central (Dowa, Mchinji, and Kasungu) and southern (Thyolo) regions in nine out of 12 sites surveyed. When microscopically examined, urediniospores were elliptical, echinulate, and hyaline to pale yellowish brown. Leaves exhibiting sporuliferous uredinia were sent by APHIS permit to the University of Illinois. To confirm the pathogen, symptomatic soybean leaf tissue of approximately 1 cm2 was excised from each of the samples, and DNA was extracted using the FastDNA Spin Kit (MP Biomedicals, Solon, OH), with further purification using the MicroElute DNA Clean-up Kit (Omega Bio-Tek, Norcross, GA). The resulting DNA was analyzed by quantitative PCR using published Taqman assays for P. pachyrhizi and P. meibomiae, with a multiplexed exogenous internal control reaction to validate negative results (1). P. pachyrhizi DNA was detected in excess of 180,000 genome equivalents/cm2 in all samples, indicating a substantial infection. P. meibomiae DNA was determined to be absent from all samples, within the limit of quantification of ~2 pg DNA/cm2. Urediniospores dislodged from three leaves and inoculated onto susceptible soybean cultivar Williams 82 produced tan lesions after 2 weeks of incubation in a detached-leaf assay. This is the first confirmed report of P. pachyrhizi causing rust on soybean in Malawi, putting at risk 14,000 ha currently under soybean production. The reports of soybean rust in Malawi and adjoining countries will alter soybean production practices and research interests. In some cases, foliar application of fungicides has increased and planting dates have been changed to avoid conditions that are most conducive for rust development. Efforts to understand the virulence and genetic diversity of the pathogen in the region are needed in order to develop and deploy resistant cultivars.

Phakopsora pachyrhizi Syd. was reported on legume hosts other than soybean in Tanzania as early as 1979 (1). Soybean rust (SBR), caused by P. pachyrhizi, was first reported on soybean in Africa in Uganda in 1996 (3), and its introduction into Africa was proposed to occur through urediniospores blowing from western India to the African east coastal areas by moist northeast monsoon winds (4). The fungus rapidly spread and was reported on soybean in South Africa in 2001, in western Cameroon in 2003, and in Ghana and the Democratic Republic of the Congo in 2007 (5). A second species causing SBR on soybean, P. meibomiae, has not been reported in Africa or elsewhere, outside of the Americas. From 2012 to 2014, symptomatic leaf samples were collected in the major soybean growing areas of the Tanzanian Southern Highlands (Iringa, Mbeya, and Ruvuma regions). Symptoms of SBR included yellowing of leaves and tan sporulating lesions. These symptoms were observed at flowering through seed maturity. From fields surveyed in 2012, 2013, and 2014, SBR was observed in 5 of 14, 7 of 11, and 14 of 31 fields, respectively. Some of the leaves sampled had up to 80% of the leaf area affected. When microscopically examined, urediniospores were elliptical, echinulate, and hyaline to pale yellowish brown. In 2014, sporuliferous uredinia were observed on leaf material collected from the Iringa and Ruvuma regions of Tanzania, and a subset of these samples was sent by APHIS permit to the University of Illinois. To confirm the pathogen, symptomatic soybean leaf tissue of approximately 1 cm2 was excised from each of the samples, and DNA was extracted using the FastDNA Spin Kit (MP Biomedicals, Solon, OH), with further purification using the MicroElute DNA Clean-up Kit (Omega Bio-Tek, Norcross, GA). The DNA was subjected to quantitative PCR using published Taqman assays for P. pachyrhizi, P. meibomiae, and a multiplexed exogenous internal control reaction to validate negative results (2). P. pachyrhizi DNA was detected in excess of 66,000 genome equivalents/cm2 in all samples, and P. meibomiae DNA was determined to be absent from all samples (limit of quantification ~2 pg DNA/cm2). Free surviving urediniospores were dislodged from 12 samples and inoculated onto susceptible soybean cultivar Williams 82, which produced sporulating SBR lesions after 2 weeks of incubation in a detached-leaf assay. Thus, Koch's postulates were completed. This is the first report of P. pachyrhizi causing rust on soybean in Tanzania. In vivo cultures have been established from most of these samples, and ongoing research includes an evaluation of the P. pachyrizi virulence on a differential set, and characterization of the genetic diversity.

Nigeria and Uganda are the closest countries to the Democratic Republic of Congo (DRC) where soybean rust caused by Phakopsora pachyrhizi has been reported. In February 2007, during a disease survey in DRC, soybean (Glycine max) leaves with rust symptoms (tan, angular lesions with erumpent sori exuding urediniospores) were observed in 10 fields in the following areas in Bas Congo Province: Bangu, Kimpese, Kolo-Fuma, Lukala, Mbanza-Ngungu, Mpalukide, Mvuazi, and Ntemo. Rust incidence in these fields ranged from 85 to 100%, while severity ranged between 3 and 35% of the leaf area on infected plants. Urediniospores were hyaline, minutely echinulate, and 23 to 31 × 16 to 20 µm. Within a week of collection, infected leaf samples were sent to the USDA-ARS Foreign Disease-Weed Science Research Unit (FDWSRU) for pathogen identification. DNA was extracted from sections of leaves containing sori with the Qiagen DNeasy Plant Mini kit (Valencia, CA), and all 10 field samples amplified in a real-time fluorescent PCR with the P. pachyrhizi-specific primers Ppm1 and Ppa2. Infected leaves of cultivar Vuangi collected from one field each in the INERA Research Station, Kimpese-Crawford, and Kimpese-Ceco were separately washed in sterile water to collect urediniospores that were used to separately inoculate three detached leaves of susceptible cultivar TGx 1485-1D. Lesions on inoculated leaves developed 5 days after inoculation (DAI), and pustules (110 to 130 µm) formed 7 DAI and erupted 2 days later exuding columns of urediniospores similar in size to the initially collected isolates. Inoculation of another set of detached leaves with a spore suspension (1 × 10(^6) spores per ml) from the first set of detached leaves resulted in typical rust symptoms. Seedlings of cultivar Williams also showed typical rust symptoms when inoculated separately with urediniospores collected from nine fields (i.e., all except Kimpese-Ceco, which was infective in the detached leaf assay). Inoculation and incubation were carried out at the FDWSRU Plant Pathogen Containment Facility at Fort Detrick as described earlier (2). The PCR assay, morphological characters of the isolates, and pathogenicity tests demonstrate that P. pachyrhizi occurs in DRC. To our knowledge, this is the first report of P. pachyrhizi infecting soybean in DRC.

Soybean rust, caused by Phakopsora pachyrhizi, is an important foliar disease of soybean. Disease severity is dependent on several environmental factors, although the precise nature of most of these factors under field conditions is not known. To help understand the environmental factors that affect disease development, soybean rust epidemics were studied in Nigeria by sequentially planting an early-maturing, highly susceptible cultivar, TGx 1485-1D, and a late-maturing, moderately susceptible cultivar, TGx 1448-2E, at 30- to 45-day intervals from August 2004 to September 2006. Within each planting date, disease onset occurred earlier on TGx 1485-1D than on TGx 1448-2E, and rust onset was at least 20 days earlier on soybean planted between August and October than on soybean planted between November and April. The logistic model provided a better description of the temporal increase in rust severity than the Gompertz model. Based on the logistic model, the highest absolute rates of disease increase were observed on soybean planted in September 2006 and October 2004 for TGx 1485-1D and TGx 1448-2E, respectively. Disease severity as measured by the relative area under disease progress curve (RAUDPC) was significantly (P < 0.05) negatively correlated with evaporation (r = –0.73), solar radiation (r = –0.59), and temperature (r = –0.64) but positively correlated with urediniospore concentration (r = 0.58). Planting date and soybean cultivar significantly (P < 0.05) affected disease severity, with severity being higher on soybean crops planted during the wet season than those planted in the dry season. Within the wet season, planting in May and July resulted in a significantly (P < 0.05) lower RAUDPC than planting between August and October. Yields were significantly (P < 0.001) related to RAUDPC during the wet season, whereby an increase in RAUDPC resulted in a linear decrease in yield. This study suggests that selection of planting date could be a useful cultural practice for reducing soybean rust.

Soybean rust, caused by Phakopsora pachyrhizi, is a major disease limiting soybean production primarily in the tropics and subtropics of Asia. Research at the Asian Vegetable Research and Development Center (AVRDC) has focused on monitoring disease development; evaluating yield losses; obtaining basic information on the biology of the fungus; and on finding sources of resistance and developing these sources into breeding lines. Rust was monitored on one moderately resistant and three susceptible lines at five locations in Taiwan during three separate seasons. Apparent infection rates were similar within lines over locations and seasons. Several experiments showed that soybean maturation was significantly positively correlated to the rate of rust development, whereas effects due to the environment and the host genotype were not as highly correlated. To compare soybean lines, methods were developed to compensate for differences in host maturities. The best method used the relative soybean life time (RLT) as a time element from 0 to 100. The time between planting and maturity was converted to a percentage of the soybean life cycle completed. Factors related to pathogenic diversity of the fungus, and the effect of environmental parameters were studied. Nine races were identified from forty-two isolates using a differential set consisting of 11 lines. The predominant races were complex with multiple virulence factors for compatibility on the differentials. Studies on leaf wetness and temperature indicated that the optimum temperature for uredospore germination was 15~ 25°C ; the minimal dew period for infection was 6 hours at 20~ 25°C and 8-10 hours at 15~ 17.5°C ; and a mean night temperature below 15°C greatly reduced lesion numbers or completely prevented lesion development. Field studies showed that precipitation was a critical factor in the development of epidemics. It was used to predict rust severity, and was more important than frequency and intensity of the infection period which consisted of leaf wetness, temperature, and their interaction. Difficulties associated with identifying and quantifying rate-reducing resistance and the ineffectiveness of race-specific resistance have brought about techniques to develop higher soybean yields with tolerance to rust. Also techniques were developed to better quantify and understand the components involved in partial resistance. In other studies, new sources of resistance were identified in accessions of the wild perennial Glycine species. (Key words: Phakopsora pachyrhizi, infection rates, perennial Glycine spp., yield losses, races)