Development of a molecular marker specific to a novel CMS line in radish (Raphanus sativus L.).
In this study, we have investigated the cytoplasmic male sterility (CMS) of a novel male sterile radish line, designated NWB CMS. The NWB CMS was crossed with 16 fertile breeding lines, and all the progenies were completely male sterile. The degree of male sterility exhibited by NWB CMS is more than Ogura CMS from the Cruciferae family. The NWB CMS was found to induce 100% male sterility when crossed with all the tested breeding lines, whereas the Ogura CMS did not induce male sterility with any of the breeding lines. PCR analysis revealed that the molecular factor that influenced Ogura CMS, the orf138 gene, was absent in the NWB CMS line, and that the orf138 gene was not also expressed in this CMS line. In order to identify the cytoplasmic factors that confer male sterility in the NWB CMS line, we carried out RFLP analyses with 32 mitochondrial genes, all of which were used as probes. Fourteen genes exhibited polymorphisms between the NWB CMS line and other radish cultivars. Based on these RFLP data, intergenic primers were developed in order to amplify the intergenic regions between the polymorphic genes. Among these, a primer pair at the 3' region of the atp6 gene (5'-cgcttggactatgctatgtatga-3') and the 5' region of the nad3 gene (5'-tcatagagaaatccaatcgtcaa-3') produced a 2 kbp DNA fragment as a result of PCR. This DNA fragment was found to be specific to NWB CMS and was not present in other CMS types. It appears that this fragment could be used as a DNA marker to select NWB CMS line in a radish-breeding program.
Nahm
Seok-Hyeon
SH
Biotechnology Institute, Nong Woo Bio Co., Yeoju, Gyeonggi, South Korea. shnahm1@nongwoobio.co.kr
Lee
Hee-Jeong
HJ
Lee
Shi-Woo
SW
Joo
Gyun-Young
GY
Harn
Chee-Hark
CH
Yang
Seung-Gyun
SG
Min
Byung-Whan
BW
D015152:Blotting, Northern; D001947:Breeding / Q000379:methods; D003433:Crosses, Genetic; D017931:DNA Primers; D004272:DNA, Mitochondrial / Q000235:genetics; D017343:Genes, Plant / Q000235:genetics*; D005819:Genetic Markers / Q000235:genetics*; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D031224:Raphanus / Q000235:genetics*; D012098:Reproduction / Q000235:genetics; D013045:Species Specificity
D017931:DNA Primers; D004272:DNA, Mitochondrial; D005819:Genetic Markers