First Report of Gaeumannomyces graminis var. graminis on Kikuyugrass (Pennisetum clandestinum) in the United States.

Kikuyugrass (Pennisetum clandestinum) is a warm-season grass and invasive weed in the landscape, but can be used for golf course fairways in southern California. In 1999, a decline of kikuyugrass was observed on golf courses in southern California beginning in late summer or early autumn. Symptoms included sunken, bleached patches of turf with individual plants having chlorotic foliage and reduced vigor. Roots and stolons were often covered with dark, ectotrophic fungi, and lobed hyphopodia were visible on the stolons. On colonized roots, the cortex was rotted, and the stele showed evidence of colonization by the fungus. In March 2002, a sample of kikuyugrass exhibiting decline symptoms was obtained from a golf course fairway in Los Angeles, CA. Sections of roots and stolons were surface sterilized for 60 s in a 0.3% sodium hypochlorite solution and placed on acidified water agar. Emerging colonies were transferred to potato dextrose agar (PDA). Isolates were characteristic of Gaeumannomyces spp. (2) with dark hyphae and curled colony edges. The rDNA internal transcribed spacer (ITS) regions of two isolates (HCC-5 and -6) were amplified by polymerase chain reaction (PCR) using universal fungal rDNA primers ITS 4 (5'-TCCTCCGCTTATTGATATGC-3') and ITS 5 (5'-GGAAGTAAAAGTCG TAACAAGG-3') (3). PCR products were sequenced and exhibited 99% sequence identity to G. graminis var. graminis (GenBank Accession No. 87685). These isolates were grown separately on autoclaved sand and cornmeal media (1) for 21 days at 25°C. Styrofoam cups were partially filled with autoclaved medium-coarse sand, and 10 g of inoculum was spread evenly in a layer on top. This layer was covered by an additional centimeter of autoclaved sand and 5 g of kikuyugrass seed (cv. 'AZ-1'). Both isolates were tested separately using six replicate cups per isolate. Controls were prepared using only a 10 g layer of autoclaved sand and cornmeal. Cups were misted at 1 h intervals on a greenhouse bench maintained at 25°C. Seeds germinated and emerged after ≈10 days. In cups inoculated with isolate HCC-5 or -6, dark mycelia were evident on the coleoptiles of the emerging plants. Plants were removed and washed 21 days after planting. Inoculated plants were chlorotic and had reduced root and foliar growth compared to the controls. Coleoptiles, hypocotyls, and roots were covered with dark, ectotrophic fungi with lobed hyphopodia present on the hypocotyls. In colonized roots, cortical tissue was rotted with extensive colonization of the epidermis and penetration of the fungus into the root cortex. Sections of infected root tissue were surface disinfested, placed on acidified water agar, and the resulting colonies transferred to PDA. Isolates exhibited the same colony morphology and characteristics as those previously identified as G. graminis var. graminis. To our knowledge, this is the first report of this fungus as a pathogen of kikuyugrass. References: (1) M. J. C. Asher. Ann. Appl. Biol. 70:215, 1972. (2) P. C. Cunningham. Isolation and culture. Pages 103-123 in: Biology and Control of Take All. M. J. C. Asher and P. J. Shipton, eds. Academic Press, London, 1981. (3) T. J. White et al. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. Pages 315-322 in: PCR Protocols: A Guide to Methods and Applications. M. A. Innis et al. eds. Academic Press, San Diego, CA, 1990.