First Report of Ophiosphaerella herpotricha Causing Spring Dead Spot of Bermudagrass in Mississippi.

Spring dead spot (SDS) is the most destructive disease of bermudagrass (Cynodon dactylon (L.) Pers.). Symptoms of SDS appear in the spring when bermudagrass transitions out of winter dormancy. These symptoms include depressed, straw-colored patches that range from several centimeters to a meter in diameter. Infected roots and rhizomes are black, brittle, and necrotic. The disease is caused by three species of fungi: Ophiosphaerella herpotricha (Fr:Fr) J. Walker; O. korrae (J. Walker & A.M. Smith) Shoemaker & C.E. Babcock; or O. narmari (J. Walker & A.M. Smith) Wetzel, Hubert & Tisserat. However, O. korrae is the most prevalent causal organism of SDS in the southeastern United States and was the only species reported in Mississippi (1). In April of 2006, root samples were collected from a bermudagrass putting green in Booneville, MS with a high level of SDS incidence and severity. Symptomatic roots were collected and surface disinfested in 0.6% NaOCl and plated on one-quarter-strength potato dextrose agar (PDA) amended with streptomycin sulfate and chloramphenicol. Hyphae growing from the roots were transferred to full-strength PDA after 5 to 7 days. Mycelium from five pure-culture colonies plus an O. herpotricha control was harvested after 4 weeks of growth and the genomic DNA was extracted. The genomic DNA of the Booneville isolates and the O. herpotricha control were amplified by PCR using species-specific primers OHITS1 and OHITS2 for O. herpotricha (2). Amplification of a 454-bp fragment of DNA confirmed one of the five unknown isolates as O. herpotricha. The other four isolates were not identified. 'Sahara' bermudagrass (4 weeks old in 3.8 × 20 cm Cone-tainers containing a sand and soil mixture) was inoculated with the Booneville-O. herpotricha isolate and the O. herpotricha control. One gram of oat seed infested with O. herpotricha isolates was inserted 2 cm below the crowns in the root zone of bermudagrass plugs. The inoculated bermudagrass plants were incubated for 4 weeks in the greenhouse. A control consisting of noninfested sterile oats was included. Following incubation, black, necrotic roots were observed on the plants inoculated with both O. herpotricha isolates. No symptoms were observed on roots of noninfested plants. Symptomatic roots were disinfested and plated on one-quarter-strength PDA. Koch's postulates were completed after O. herpotricha was reisolated from roots of plants inoculated with both O. herpotricha isolates and confirmed by PCR as mentioned above. The identification of O. herpotricha as a causal organism of SDS in Mississippi clarifies the involvement of multiple causal agents in this state and broadens the geographic distribution of this root-rot species. References: (1) F. B. Iriarte et al. Plant Dis. 88:1341, 2004. (2) N. A. Tisserat et al. Phytopathology 84:478, 1994.