First Report of Leaf and Sheath Spot Caused by Waitea circinata var. zeae on Paspalum vaginatum and Zoysia tenuifolia in China.

Abstract

Waitea circinata var. zeae was previously reported as the causal agent of leaf and sheath spot on Festuca arundinacea (1) and Panicum tennesseense (2) in the United States. In late May to mid-September 2013, a disease resembling leaf and sheath spot was observed on Paspalum vaginatum in fairways from several golf courses in Hainan Province, China. Affected plants initially had large yellow and brown spots on leaves and sheathes, then the whole plant turned yellowish-brown and eventually died. The same symptoms were also observed on a lawn established with Zoysia tenuifolia in Hainan University. Symptomatic leaves were surface sterilized in 1% hypochlorite for 1 min, rinsed with sterile water three times, and plated onto potato dextrose agar (PDA) amended with 0.01% gentamicin sulfate. Two Rhizoctonia-like fungal isolates were obtained from the diseased P. vaginatum (Isolate no. ML-WC1) and Z. tenuifolia (Isolate no. HNU-1) samples. After incubation on PDA for 1 week at 25°C, white mycelial colonies developed and eventually turned a salmon color with age. Small, white, spherical sclerotia (0.5 to 1 mm in diameter) were observed submerged throughout the agar media after incubation for 1 week and turned a reddish-brown color within 4 weeks. The two isolates were tentatively identified as W. circinata var. zeae based on these characteristics. The internal transcribed spacer (ITS) region of the ribosomal DNA gene was amplified and sequenced using primer pair ITS1/ITS4. The sequences of the two isolates (GenBank Accession Nos. KJ717943 and KJ717944) were more than 99% similar to W. circinata var. zeae (JQ688056 and JQ688059) sequences deposited in GenBank. To confirm pathogenicity, inocula were prepared by incubating autoclaved rye grains with strains ML-WC1 and HNU-1 for 2 weeks at 25°C. Ten colonized rye grains were uniformly spread around the crowns of 6-week-old P. vaginatum and Z. tenuifolia seedlings grown in 10-cm-diameter pots. Each isolate was placed in four separate pots and four control plants were treated with non-inoculated grain. All pots were covered with translucent plastic bags and placed in a greenhouse at 24 ± 2°C with a 12-h light/dark cycle. By 1 week post-inoculation, significant blighting of leaves and sheaths was observed, while non-inoculated plants showed no symptoms. W. circinata var. zeae was successfully re-isolated from symptomatic plants and confirmed by its Rhizoctonia-like mycelium and small, reddish-brown, spherical sclerotia on the PDA. Recently a related species, W. circinata var. circinata, causing brown ring patch on two cool-season grasses was reported in China (3). However, the isolates of W. circinata var. zeae were distinguished from W. circinata var. circinata base on ITS sequence data and morphological characters. To our knowledge, this is the first report of W. circinata var. zeae infecting P. vaginatum and Z. tenuifolia in China.