Reid D. Frederick

Epidemics of Soybean Rust (Phakopsora pachyrhizi) in Brazil and Paraguay from 2001 to 2003

In 5 March 2001, a severe rust outbreak was recorded at Pitapó, Paraguay, and the causal organism was determined to be Phakopsora pachyrhizi using polymerase chain reaction (PCR) and DNA sequence analysis. In May, rust surveys showed spread throughout most of Paraguay and into western and northern Parana, Brazil. In the 2001-02 season, rust was widespread in Paraguay, but losses were reduced due to severe drought; however, in Brazil it spread to more than 60% of the soybean acreage, causing field losses estimated at 0.1 million metric tons (MMT). In 2003, the disease was observed in more than 90% of the fields in Brazil, and the projected losses in Mato Grosso and Bahia alone are 2.2 MMT (US

Real-time PCR and its application for rapid plant disease diagnostics

487.3 million). Approximately 80% of the soybean acreage in Brazil was sprayed twice with fungicides at the cost of US

Breeding for resistance to soybean rust

544 million. Differences in efficacy have been observed among the commercial strobilurin and triazol fungicides.

Evaluation of virulence of Phakopsora pachyrhizi and P. meibomiae isolates.

Rapid-cycle real-time polymerase chain reaction (PCR) methods may revolutionize the manner in which plant pathogens are identified and diseases are diagnosed. As the genomics age progresses and more and more DNA sequence data become available, highly specific primers and fluorescent probe sequences can be designed to yield target amplicons to unique regions of a pathogens genome. Portable real-time PCR instruments described here are now allowing for diagnostic assays to be run directly in the field or at remote locations other than the standard diagnostic laboratory. Rapid real-time PCR diagnosis can result in appropriate control measures and (or) eradication procedures more quickly and accurately than traditional methods of pathogen isolation. Disease losses are minimized and control costs reduced. Advantages and disadvantages of rapid real-time PCR for the detection of bacterial, fungal, and viral plant pathogens are described.

Polymerase Chain Reaction Assays for the Detection and Discrimination of the Soybean Rust Pathogens Phakopsora pachyrhizi and P. meibomiae

Soybean rust occurs in all major soybean-growing regions of the world including the North American mainland. Soybean rust, caused by Phakopsora pachyrhizi, is the most destructive foliar disease of soybean, and yield losses of over 50% are common when environmental conditions are conducive for disease development. Heavily infected plants defoliate and mature more rapidly than plants not infected with rust. P. pachyrhizi has a broad host range and can infect many other legumes including some native to Australia. A number of physiological races of the fungus have been reported on these native legumes from Australia and on soybean. In addition, four single genes for rust resistance were previously identified in four different soybean plant introductions. These sources of resistance also have been reported to be susceptible in some field locations and when challenged with certain isolates of P. pachyrhizi. Partial resistance, expressed as reduced pustule number and increased length of latent period, has also been reported but has not been widely used in breeding programs. Yield stability has been used in the past and compares percentage of yields in fungicide and nonfungicide plots. Cultivars or lines with a higher percentage of yield have greater yield stability in the presence of rust. Although soybean rust only recently was found in the continental United States, a proactive project to evaluate the USDA soybean germ plasm collection for rust resistance was initiated in 2002 at the Fort Detrick plant biocontainment facility and at six international locations. Part of this project is to discover soybean lines with greater yield stability, and additional single and partial resistance. To help minimize the impact of soybean rust, the first line of defense will be fungicides, with host resistance and yield stability augmenting the long-term management of soybean rust.

International Fungicide Efficacy Trials for the Management of Soybean Rust

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.

Differential responses of resistant soybean entries to isolates of Phakopsora pachyrhizi

ABSTRACT Soybean rust occurs in Australia and many countries throughout Africa, Asia, and South America. The causal agents of soybean rust are two closely related fungi, Phakopsora pachyrhizi and P. meibomiae, which are differentiated based upon morphological characteristics of the telia. Determination of the nucleotide sequence of the internal transcribed spacer (ITS) region revealed greater than 99% nucleotide sequence similarity among isolates of either P. pachyrhizi or P. meibomiae, but only 80% sequence similarity between the two species. Utilizing differences within the ITS region, four sets of polymerase chain reaction (PCR) primers were designed specifically for P. pachyrhizi and two sets for P. meibomiae. Classical and real-time fluorescent PCR assays were developed to identify and differentiate between P. pachyrhizi and P. meibomiae. Identification of P. pachyrhizi from infected soybean leaves using the real-time PCR assay will allow for more rapid diagnoses.

Identification of a new soybean rust resistance gene in PI 567102B

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.

Adult Plant Evaluation of Soybean Accessions for Resistance to Phakopsora pachyrhizi in the Field and Greenhouse in Paraguay

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.

New Legume Hosts of Phakopsora pachyrhizi Based on Greenhouse Evaluations

Soybean rust (SBR) caused by Phakopsora pachyrhizi Syd. and P. Syd. is one of the most economically important diseases of soybean (Glycine max (L.) Merr.). Durable resistance to P. pachyrhizi is the most effective long-term strategy to control SBR. The objective of this study was to investigate the genetics of resistance to P. pachyrhizi in soybean accession PI 567102B. This accession was previously identified as resistant to SBR in Paraguay and to P. pachyrhizi isolates from seven states in the USA (Alabama, Florida, Georgia, Louisiana, Mississippi, South Carolina, and Texas). Analysis of two independent populations, one in which F2 phenotypes were inferred from F2-derived F3 (F2:3) families and the other in which F2 plants had phenotypes measured directly, showed that the resistance in PI 567102B was controlled by a single dominant gene. Two different isolates (MS06-1 and LA04-1) at different locations (Stoneville, MS and Ft. Detrick, MD) were used to independently assay the two populations. Linkage analysis of both populations indicated that the resistance locus was located on chromosome 18 (formerly linkage group G), but at a different location than either Rpp1 or Rpp4, which were previously mapped to this linkage group. Therefore, the SBR resistance in PI 567102B appeared to be conditioned by a previously unreported locus, with an underlying single dominant gene inferred. We propose this gene to be designated Rpp6. Incorporating Rpp6 into improved soybean cultivars may have wide benefits as PI 567102B has been shown to provide resistance to P. pachyrhizi isolates from Paraguay and the US.