First Report of Root Rot on Atractylodes macrocephala (Largehead Atractylodes Rhizome) Caused by Ceratobasidium sp. in China

Atractylodes macrocephala is a perennial herbaceous plant (family Asteraceae) native to China. The biennial root, Largehead Atractylodes Rhizome (LAR), is the most commonly used Chinese herbal medicine to prevent early pregnancy loss due to miscarriage. From summer 2010 to spring 2012, symptoms of root rot were observed on LAR in Xianfeng county, Enshi city, Hubei Province, China. White mold on the root of LAR could be observed at an early growth stage in the field and the white mold spread over the entire plant after 10 days, which differs from root rot of LAR caused by Fusarium oxysporum and Rhizoctonia solani, neither of which are characterized as having mycelium spreading over the whole plant (4). Where root rot symptoms were present, rhizome yield was reduced by 15% on average, with up to 40% yield loss in some fields. Under humid conditions in mid-June, the disease in the field spread quickly and the rhizomes of LAR were completely rotted. After rainfall and increasing temperature from 16 to 35°C, white mycelium appeared and plants withered within a few weeks. In April 2011 and 2012, a fungus was consistently recovered from symptomatic rhizome samples after they were surface sterilized with 0.1% mercuric chloride solution and plated onto potato dextrose agar (PDA). Pale gray colonies with short aerial mycelia and brown sclerotia formed on PDA after 7 days incubation at 28°C. Binucleate cells were observed using light microscopy and the characteristics were matched with morphological characteristics of a Ceratobasidium sp (3). Genomic DNA of the culture was extracted, and the rDNA-internal transcribed spacer sequence (GenBank Accession No. JQ926741) showed 99% identity to Ceratobasidium sp (GenBank No. H269825.1). Mycelial plugs of the culture taken from PDA were inoculated onto 40 rhizomes of 1-year-old seedlings and plants were incubated with a 16-h photoperiod at 28°C and 90% relative humidity in an artificial climate chamber where they developed typical disease symptoms after 2 days. Ten rhizomes of 1-year-old seedlings and were treated with PDA plugs only. All seedlings inoculated with the pathogen were withered and the rhizomes were completely covered with gray mycelium 2 days after inoculation, which was similar to the symptoms observed in the field. After 7 days, the symptoms were more severe than those observed in the field, with seedlings rotted completely. The main stalk of all inoculated plants was covered with gray mycelia in 4 days, and