Abstract
Peanut, Arachis hypogaea L., is an important source of vegetable oil and protein in China with a planting area of 5 million ha and annual production of 16.2 million metric tons. In September of 2012, typical symptoms of Sclerotinia blight were first observed on peanut in Shuangcheng, Heilongjiang Province, China. Approximately 50% of the plants in a field were infected. Symptoms began as a chlorotic wilt on the foliage and developed into necrosis of basal stems and developed very quickly in the field. In advanced stages of the disease, stems and branches became bleached and eventually died. White, fluffy mycelium and black irregular sclerotia (3.5 to 5.4 mm diameter) were observed on the infected stems. Infected branches and pegs were shredded, and most pods dropped on the soil during harvest. To isolate the causal agent of the disease, sclerotia were collected from the field. Twenty sclerotia were surface disinfected in 1% NaOCl for 3 min, rinsed three times with sterile water, placed on potato sucrose agar (PSA) with 100 μg/ml streptomycin, and then incubated at 22°C in the dark for 10 days. Fungal DNA was extracted from mycelia with a TIANGEN DNAsecure Plant Kit (Beijing) and amplified by PCR with the universal fungal primer ITS1 and ITS4. PCR products of five replicates were sequenced and subjected to an NCBI BLAST search. The BLAST search revealed that our sequences (GenBank Accession No. KC935388) had 100% identity with reported sequences of Sclerotinia sclerotiorum. The isolates were identified as S. sclerotiorum (Lib.) de Bary based on mycelia, sclerotia, and rDNA sequence analysis. To conduct pathogenicity tests, nine potted peanut plants (1 month old) were each inoculated with a 5-mm-diameter disk of colonized PSA by placing the inoculum on the base of the stem. Plants were inoculated with a plug of non-colonized PSA as controls. All inoculated plants were covered with a plastic bag for 5 days to maintain high humidity and incubated at 22 to 24°C in the growth chamber with a 12-h photoperiod. After 2 days, the inoculated plants showed water-soaked brown symptoms on the stem base and whole plants wilted after 5 days under high moisture conditions. The symptoms were identical to those observed on peanut plants in Shuangcheng, Heilongjiang Province, whereas the control plants remained symptom-free. Re-isolation of the fungus from the inoculated plants confirmed that the causal agent was S. sclerotiorum. S. sclerotiorum has been reported on peanut in the United States and Argentina (1,2,3). To our knowledge, this is the first report of S. sclerotiorum as a pathogen causing Sclerotinia blight on peanut in northeastern China.
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