First report of Ophiosphaerella narmari causing spring dead spot of hybrid bermudagrass in China.

Abstract

Hybrid bermudagrass (Cynodon dactylon×C. transvaalensis) is widely used as turf in transition zone of China. Spring dead spot (SDS) is one of the most damaging diseases of hybrid bermudagrass. Symptoms of SDS appear when hybrid bermudagrass starts to break dormancy with warm temperature in early spring. The symptoms show sunken, circular or irregularly shaped, straw-colored patches, with 20 to 100 cm in diameter. The patches maintain dormant as the surrounding uninfected turfgrass resumes growth and turns green. SDS pathogens are soilborne fungi that colonize roots, stolons and rhizomes, infected roots or rhizomes become black and eventually collapse. Three species of fungi are reported to cause SDS: Ophiosphaerella herpotricha (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 (Walker and Smith 1972; Walker 1980; Shoemaker and Babcock 1989; Wetzel et al. 1999). However, distribution of the three species may vary by geographical region (Cottrill et al. 2016). In October 2020, symptoms of SDS were observed on hybrid bermudagrass fairways of Taihu golf course in Wuxi, Jiangsu province. Root samples of SDS were collected, symptomatic roots with 3-4 cm length were cut, washed 2-3 times, surface sterilized in 0.6% NaOCl for 5 min, rinsed and blotted dry for 2 min and placed on potato dextrose agar (PDA) amended with 50 mg L-1 each of ampicillin, streptomycin sulfate and tetracycline. Plates were incubated in the dark at 25℃ for 5-7 days, Hyphae growing from the roots were transferred to new PDA plates. A total of 7 fungal isolates with morphology similar to SDS pathogens were obtained (Tredway et al. 2009). The genomic DNA was extracted from 2 of them (7-41, 8-6) and amplified with universal primers ITS5 and ITS4 (White et al. 1990). PCR products were sequenced (deposited as MW536995 and MW536994 in GenBank, not available yet) and showed 99.79% similarity to O. narmari (KP690979). Pathogenicity tests were performed on 'Tifdwarf' hybrid bermudagrass (9-week-old in 5 × 20 cm Cone-Tainers containing a sand and nutrition substrate mixture). Eight oat seeds infested with O. narmari were inserted 5 cm below the soil surface in the root zone of hybrid bermudagrass. The inoculated turfgrass grew for five weeks in the growth chamber with a 12-h day/night cycle of 25/20°C and 90% relative humidity. A control treatment was inoculated with 8 noninfested sterile oat seeds, and each treatment was replicated 3 times. The root tissues of hybrid burmudagrass inoculated with O. narmari became black and necrotic, no symptoms were observed on the roots of noninfested plants. O. narmari was consistently reisolated from symptomatic roots, and confirmed by PCR as mentioned above. To the best of our knowledge, this is the first report of O. narmari caused spring dead spot in the transition zone of China. The identification of SDS caused by O. narmari will have important implications for the management of this root-rot species on hybrid bermudagrass.