Characterization of CMS and maintainer lines in indica rice (Oryza sativa L.) based on RAPD marker analysis

Total DNA from WA type CMS lines: Zhenshan 97 A, Longtepu A and their maintainers Zhenshan 97 B, Longtepu B was extracted by CTAB method. One hundred primers were used for screening RAPD markers to distinguish CMS line (A) and maintainer (B) plants at seedling stage. Results showed that under the conditions of 37 °C annealing temperature and 1.5 mM MgCl₂ concentration, in Zhenshan 97 A, Longtepu A there was a 1600 bp DNA fragment in product amplified by primer OPA12, while in Zhenshan 97 B, and Longtepu B no 1600 bp fragment was found. The 1600 bp fragment was also found in DA type CMS line Xieqingzao A, but was absent in Xieqingzao B. Also in the restorer line, Minghui 63 the 1600 bp fragment was absent. In F₁ and F₂ generation of Zhenshan 97A/Minghui 63, all plants investigated had the 1600 bp fragment. When mitochondrial DNA (mtDNA) was isolated from the three CMS (A) and their B lines and amplified by OPA12, results showed that the 1600 fragment was found in all the three A lines and was absent in the three B lines. In DA type Xieqingzao A, two other fragments (700 bp, 1000 bp) were also found except the 1600 bp. These results indicate that the 1600 bp fragment derived from CMS mitochondrial DNA can be used as a RAPD marker to distinguish A and B plants at seedling stage, and the fragments (700 bp, 1000 bp) can be used to distinguish WA and DA cytoplasms.