RAPD Analysis Of Mitochondrial DNAs Of Cytoplasmic Male Sterile Line And Its Maintainer In Tobacco

Tobacco is an economic crop of great importance. In modern tobacco agricultural production, taking advantage of cytoplasmic male sterile (CMS) lines and hybrid varieties have become the main development direction. In this study, mitochondnal DNA from 2 male sterile lines and their maintain lines of tobacco cultivars were extracted. We analyze the difference between male sterile lines and maintainer lines on nucleoside acid by RAPD method. Resequencing was applied to compare the difference between cytoplasmic male sterile line and its maintainer on gene. The results are as follows:1. The RAPD of mitochondrial DNAThe mitochondrial DNA of male sterile lines and maintainer lines in tobacco K326, MSK326, K394 and MSK394 were analysed by the method of random amplified polymorphic DNA using 100 random 10-base primers. The result showed that 5 primers did not amplified any band and 86 primers,90.53% of total primers, did not amplified polymorphic band between CMS and maintainer lines. This indicated the homolog of mitochondria between CMS and maintainer line. Mean while,9 primers amplified polymorphic band between CMS and maintainer lines. The differences may indicated some relationship between cytoplasmic male sterility.2. The contrast of genetic coding sequences of target genesFour selected materials were tested by resequencing method and the results were contrasted to genetic coding sequences of mitochondrial DNA genes (GenBank: BA000042).We found that different genetic coding sequences exist in nad6, sdh4, coxl, atp4, atp6, atp8, orf239, orfl08, orf129, orf147, orf306, orf315, orf144, orf118, orf177, orf274, orf155 and orf161 gene between male sterile lines and maintainer lines in tobacco. And bases mutation led to amino acid mutation.