MOLECULAR AND BIOLOGICAL STUDIES ON MALE-STERILE CYTOPLASM IN THE CRUCIFERAE .1. THE ORIGIN AND DISTRIBUTION OF OGURA MALE-STERILE CYTOPLASM IN JAPANESE WILD RADISHES (RAPHANUS-SATIVUS L) REVEALED BY PCR-AIDED ASSAY OF THEIR MITOCHONDRIAL DNAS

Ogura male-sterile cytoplasm was surveyed in common Japanese radish cultivars and in wild radishes growing in various localities in Japan. Mitochondrial (mt) DNA rearrangement involving the atp6 gene was used as a molecular marker. To detect the mtDNA rearrangement, polymerase chain reactions (PCR) were designed to amplify the upstream region of the atp6 gene. The oligonucleotides homologous to the following three regions were synthesized: (1) trnfM, (2) ORF105 and (3) atp6. PCRs were conducted with a pair of the first and the third primers to detect normal mtDNA, and with the second and the third primers for Ogura-type mtDNA. All 15 Japanese cultivars yielded an amplification product which was the same as that of normal mtDNA, whereas some wild radishes gave the product specific to Ogura mtDNA. Twenty-four populations of wild radish were classified into three groups according to the frequency of Ogura-type mtDNA: (1) in ten populations, all four plants analyzed per population had normal type mtDNA, (2) in five populations, only plants with Ogura-type mtDNA were found, and (3) nine populations included both normal and Ogura-type mtDNAs. There were no geographical restrictions and no dine in the distribution of the plants with Ogura-type mtDNA. These results suggested that the Ogura-type; male-sterile cytoplasm originated in wild radishes.