To investigate host specialization in Macrophomina phaseolina, the fungus was isolated from soybean, corn, sorghum, and cotton root tissue and soil from fields cropped continuously to these species for 15 years in St. Joseph, LA. Chlorate phenotype of each isolate was determined after growing on a minimal medium containing 120 mM potassium chlorate. Consistent differences in chlorate sensitivity were detected among isolates from different hosts and from soil versus root. To further explore genetic differentiation among fungal isolates from each host, these isolates were examined by restriction fragment length polymorphism and random amplified polymorphic DNA (RAPD) analysis. No variations were observed among isolates in restriction patterns of DNA fragments amplified by polymerase chain reaction covering the internal transcribed spacer region, 5.8S rRNA and part of 25S rRNA, suggesting that M. phaseolina constitutes a single species. Ten random primers were used to amplify the total DNA of 45 isolates, and banding patterns resulting from RAPD analysis were compared with the neighbor-joining method. Isolates from a given host were genetically similar to each other but distinctly different from those from other hosts. Chlorate-sensitive isolates were distinct from chlorate-resistant isolates within a given host. In greenhouse tests, soybean, sorghum, corn, and cotton were grown separately in soil infested with individual isolates of M. phaseolina that were chosen based on their host of origin and chlorate phenotype. Root colonization and plant weight were measured after harvesting. More colonization of corn roots occurred when corn was grown in soil containing corn isolates compared with isolates from other hosts. However, there was no host specialization in isolates from soybean, sorghum, or cotton. More root colonization in soybean occurred with chlorate-sensitive than with chlorate-resistant isolates.

Asian soybean rust, caused by Phakopsora pachyrhizi Sydow, has been known to occur in the eastern hemisphere for nearly a century. More recently, it was reported from Hawaii in 1994, eastern and southern Africa from 1996-1998, Nigeria in 2001, and Brazil and Paraguay in 2002. Aerobiological models suggested that urediniospores of the pathogen would be disseminated on wind currents to the continental United States in association with tropical storms if the disease became established north of the equator during hurricane season (U.S. Soybean Rust Detection and Aerobiological Modeling online publication at www.aphis.usda.gov/ppq/ ep/soybean_rust/ ). Since soybean rust was observed at approximately 5°N latitude in South America before several hurricanes impacted the continental United States in September 2004, it seems likely that the introduction was associated with at least one of these tropical storms, especially hurricane Ivan. Symptoms of the disease were first observed on soybean (Glycine max (L.) Merr.) in the continental United States on November 6, 2004 in a field near Baton Rouge, LA. Typical pustules and urediniospores on infected leaves were readily apparent when viewed with a dissecting microscope. Urediniospores were obovoid to broadly ellipsoidal, hyaline to pale yellowish brown with a minutely echinulate thin wall, and measured 18 to 37 × 15 to 24 μm. Paraphyses were cylindric to clavate and slightly thickened at the apex, colorless to pale yellowish brown, and 25–50 × 6–14 μm in size. This morphology is typical of Phakopsora pachyrhizi and P. meibomiae, a less aggressive, western hemisphere species (2). DNA was extracted from leaves containing sori using the Qiagen DNeasy Plant Mini kit. P. pachyrhizi was detected using a real-time polymerase chain reaction (PCR) protocol (1) that differentiates between P. pachyrhizi and P. meibomiae performed in a Cepheid thermocycler with appropriate positive and negative controls. The PCR master mix was modified to include OmniMix beads (Cepheid). The field diagnosis of P. pachyrhizi was confirmed officially by the USDA/APHIS on November 10, 2004, and this was followed on November 11, 2004 by a wide-ranging survey of soybean and kudzu (Pueraria sp.) in soybean production areas in southern and central Louisiana. Collections from this survey also were assayed as described above, and six soybean specimens from five sites were confirmed positive. The disease was not found on kudzu samples. To our knowledge, this is the first report of P. pachyrhizi in the continental United States. Voucher specimens have been placed in the USDA National Fungus Collection. References: (1) R. D. Frederick et al. Phytopathology 92:217, 2002. (2) Y. Ono et al. Mycol. Res. 96:825, 1992.

Soybean [Glycine max (L.) Merr.] resistance to soybean rust (SBR) caused by Phakopsora pachyrhizi could reduce reliance on fungicides to manage this disease. The objective of this study was to identify soybean germplasm with resistance to field populations of P. pachyrhizi in the United States. Field evaluations of 576 accessions from the USDA Soybean Germplasm Collection for resistance to SBR were conducted at seven locations in the southern United States between 2006 and 2008. Accessions from maturity groups (MG) 000 to X and North American susceptible check cultivars from each MG except X were rated for disease severity in all year–location environments, and for disease incidence, fungal sporulation, lesion type, and/or uredinia density in certain environments. While none of the accessions was immune in all environments, 64 were resistant in two or more locations each year that they were tested. Some accessions appeared to be more resistant in certain environments than in others. Of the original four Rpp genes described in the literature, Rpp1 provided the highest level of resistance, and among the accessions with uncharacterized Rpp genes, PI 567104B had the highest overall resistance across environments. The plant introductions confirmed to be resistant in these evaluations should be useful sources of genes for resistance to North American populations of P. pachyrhizi

Verticillium wilt caused by race 2 of Verticillium dahliae is common in proportions) was about 5.7 ms per gram of, soil. In five other fields, California on tomato cultivars with the Ve gene for resistance to race 1. however, in which the numbers of race 2 ms were determined, a linear About 47% of 124 isolates of V. dahliae taken directly from tomato field correlation was observed between numbers of race 2 ms (0.0 to 2.0 ms/g of 'soils were race 2; the remaining isolates were either race 1 (43%) or soil) and DI (0 to 100%) when data were plotted arithmetically after nonpathogenic on tomato (about 10%). In contrast, isolates from diseased conversion of DI %to loge (1/ 1-DI) (slope = 2.Oand r =0.877). The line for tomato plants (race I resistant cultivars) from the same fields were the same data plotted on a logio-loge-logio scale had a slope of 1.0(or 1.57 predominantly race 2 (about 86% of 153 tested). Race I was more virulent if assumed to be nonlinear and transformed to logio before regression than race 2 on cultivars lacking the Ve gene for resistance (susceptible). analysis) instead of 0.66, as predicted by Baker et al (10) for abstract Average virulence of race 2 isolates was lower on cultivars with the Ve gene mathematical Model II. Thus, the models and equivalent interpretations than on susceptible cultivars. Incidence of Verticillium wilt (DI) on race I for slopes of lines in arithmetic, logio-logio and log0o-log,-0ogio plots appear resistant cultivars was essentially 100% in 46 fields where soil inoculum to be of questionable validity. density (ID) of total microsclerotia (ms) (race 1 and 2 in undetermined

Thirty-nine isolates of Fusarium oxysporum were collected from tomato plants displaying wilt symptoms in a field in California 2 years after F. oxysporum f. sp. lycopersici race 3 was first observed at that location. These and other isolates of F. oxysporum f. sp. lycopersici were characterized by pathogenicity, race, and vegetative compatibility group (VCG). Of the 39 California isolates, 22 were in VCG 0030, 11 in VCG 0031, and six in the newly described VCG 0035. Among the isolates in VCG 0030, 13 were race 3, and nine were race 2. Of the isolates in VCG 0031, seven were race 2, one was race 1, and three were nonpathogenic to tomato. All six isolates in VCG 0035 were race 2. Restriction fragment length polymorphisms (RFLPs) and sequencing of the intergenic spacer (IGS) region of rDNA identified five IGS RFLP haplotypes, which coincided with VCGs, among 60 isolates of F. oxysporum from tomato. Five race 3 isolates from California were of the same genomic DNA RFLP haplotype as a race 2 isolate from the same location, and all 13 race 3 isolates clustered together into a subgroup in the neighbor joining tree. Collective evidence suggests that race 3 in California originated from the local race 2 population.