Use of Mitochondrial DNA Polymorphisms to Distinguish Cytoplasms of Cultivated and Wild Species in Section Cepa of Allium.

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  • ミトコンドリア DNA 多型のネギ属 Cepa 節栽培種および野生種の細胞質判別への利用

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Abstract

Two methods for distinguishing mitochondrial DNA (mtDNA) among two cultivated species, Allium fistulosum and A. cepa, and four wild species, A. altaicum, A. galanthum, A. oschaninii and A. vavilovii, in section Cepa of Allium were compared. One method was the RFLP analysis using probes developed from mtDNA of A. cepa and the other was the PCR-RFLP analysis of two mtDNA regions, V7 region of small ribosomal subunit RNA (srRNA) gene and subunit 3 of NADH dehydrogenase (nad3)-ribosomal protein S12 (rps12) region. The same results were obtained in hybridization of probes with mtDNA and total DNA, so that the latter was used for distinguishing the mtDNA in the first method. Mitochondrial DNA polymorphisms were detected in all species combinations by RFLP analysis by using the probes. Polymorphisms were also detected in several species combinations by PCR-RFLP analysis of the V7 region of srRNA gene although they were not detected in any species combinations by analyzing the nad3-rps12 region. The two methods were effective in identifying mtDNA of reciprocal F1 hybrids between A. galanthum and A. fistulosum. In this study, effective combinations of probes and restriction enzymes and those of primers and restriction enzymes to distinguish mtDNA were successfully established for every species combination. Our results can be applied for confirmation of cytoplasmic substitution between two cultivated and four wild species in section Cepa.

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