ldentification of RAPD and SCAR Markers Linked to a Restorer Gene for Ogura Cytoplasmic Male Sterility in Radish (Raphanus sativus L.) by Bulked Segregant Analysis

概要

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DNA markers linked to a fertility restorer (Rf) gene for Ogura cytoplasmic nrale sterility in radish (Raphanus sativus L.) were screened by bulked segregant analysis using random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) and high-resolution agarose gel electrophoresis. Ten male-fertile and 10 male-sterile plants arbitrarily selected from the F_2 segregating population made by the selfing of an F_1 plant between `MS-Gensuke' (a male-sterile line possessing Ogura cytoplasm) and `Comet' (a restorer having a nuclear Rf gene), were used as the materials for bulked DNA samples. Using 10 out of 260 arbitrary prixners tested, RAPD fragments specific to the bulked DNA samples from male-fertile F_2 plants were identified. Further Itnkage analysis performed with 89 individuals in the F_2 segregating population showed that the seven RAPD markers were linked to the Rf gene. RAPD markers distributed on both sides of the Rf gene, and the closest marker, OPH11_<410> positioned 1.2 cM from the Rf gene. The three closest RAPD markers to the Rf gene were converted to SCAR markers by molecular cloning and nucleotide sequencing. This allowed identification of more reliable DNA markers linked to the Rf gene than the RAPD fragments.
日本育種学会の論文