A Stachybotrys chartarum isolate from soybean

As part of our effort to investigate fungi associated with soybean roots, Stachybotrys chartarum was isolated from soybean root lesions. Since this fungus has not been reported to cause a disease of soybean, the objectives were to identify and characterize this fungus using biological, chemical, and molecular approaches. Fungal morphology was examined using light and environmental scanning electron microscopy. Phialides bearing conidia arose from determinate, macronematous, dark olivaceous conidiophores. The phialides were obovate or ellipsoidal in whorls. Conidia were unicellular, round or ellipsoidal, 5-13 x 4-7 microm, initially hyaline with smooth walls then dark brown to black and rough-walled when mature. Radial growth of the fungus on cornmeal, oatmeal and potato dextrose agar was 38, 47, and 33 mm in diam., respectively, after 10 days at 25 degrees C. Pathogenicity was performed using sorghum grain colonized by S. chartarum placed below sown soybean seeds in a soil: sand (1:1) steam-pasteurized mix. Three weeks after inoculation, root lesions ranged from 7 to 25 mm long. The fungus was reisolated from soybean root lesions and was reidentified as S. chartarum. Biochemical analysis indicated that this soybean isolate produced satratoxins G and H along with roridin L-2, as well as the spircyclic lactones and lactams in rice culture. PCR using a S. chartarum-specific primer StacR3 and IT51 amplified a 198-bp DNA fragment from the total genomic DNA. The DNA sequence of the ITS region was 100% identical to the S. chartarum strain ATCC 9182, one nucleotide mismatch with S. chartarum strain UAMH 7900, and differed from all published sequences of 12 other species of Stachybotrys and 2 species of Memnoniella in GenBank with genetic divergence ranging from 5.26 to 9.98%. This molecular evidence further supports the identification of S. chartarum isolated from soybean root lesions.