Molecular Marker And Cytology And Differencial Display For Cytoplasmic Male Sterility (CMS) In Radish (Raphanus Sativus L.)

Radish (Raphanus sativus L.) is one of the most important vegetable crops, it shows remarkable heterosis, producing hybrid seed using Cytoplasmic Male Sterile(CMS) line as female parent is a very important way to utilize the heterosis in radish. In this study, screening the Restore fertility(Rf) gene, hybrid the CMS line and the variety with Rf, using the DDRT-PCR in the hybrid and female parent; constructing the RAMP system in radish, using it in radish CMS; light-microscopy, electro-microscopy and DDRT-PCR were performed to detect the mechanism of CMS in radish maked by many CMS line and maintainer line.The total DNA were isolated from young radish leaves, two DNA fragments about 780bp and 280bp were amplificated in primer to orf138 gene region, the two fragments exit or not simultaneity in radish. After recovering the 780bp fragment in maintainer line B₁ and CMS line A₁ and A₅, blast result shows that the three fragments is 100% homology with the reported orf138 nucleotide sequence, separately. There is no difference among the three fragments, they are 97.75% homology each other. Screening the Rf gene though the primer to STS marker linked to Rf gene, the Rf gene about fragment 190bp was discovered in "Dong fang hei da gen" only. The sterility was influenced by environment, the fertility may be changed in CMS line and maintainer line. The fragments about 780bp, 280bp and 190bp exiting or not in the radish which fertility change is same as those fertility is common. Maybe the factor related to pollen has changed. Hybrid the CMS line and "Dong fang hei da gen", after comparing the hybrid and female parent though DDRT-PCR, the difference bands exit in female parent mostly, the homology is high between the RNA in plant.The Random Amplified Microsatellite Polymorphism (RAMP) system suitable to radish was constructed. Genome DNA of Radish was analyzed by optimizing the concentration of Mg²⁺, dNTP and primer in. Three different concentrations level were set respectively, Mg²⁺ 0.75, 1.5, 3.0mmol · L^-1; dNTP 0.05, 0.15, 0.3 mmol· L^-1; primer 0.065, 0.2, 0.4 μmol· L^-1, and the suitable annealing temperature was also screened. The...