リボソームDNA-ITS領域のNested PCRによるネギ黒腐菌核病菌（Sclerotium cepivorum Berkeley）の検出

抄録

The authors demonstrated sensitive detection of allium white rot pathogen (Sclerotium cepivorum Berkeley) using a nested PCR method for the ITS region of ribosomal DNA. A specific primer pair (SCITSF2 and SCITSR4) was designed for the consensus sequences lying on ITS1 and ITS2 among 5 isolates obtained from Shizuoka prefecture and 11 isolates of S. cepivorum. No cross reactions were observed in the PCR using SCITSF2 and SCITSR4 with any of the closely related non-target 10 fungal species of Sclerotiaceae (e.g. 3 Botrytis, Ciboria, Dumontinia and 5 Sclerotinia species) and Sclerotium rolfsii. Nested PCR in combination with primer pairs for the ITS region (OLPF1 and OLPR) and SCITSF2 and SCITSR4 made it possible to detect soil infestation with S. cepivorum, and asymptomatic infections of welsh onion with S. cepivorum, whereas the PCR using SCITSF2 and SCITSR4 partially failed to detect them. These results indicated that this method is capable of specific and sensitive detection of white rot disease infection in culture fields.