Establishment Of MtDNA Fingerprinting Of CMS Lines And Maintainer Lines Of Soybean By RAPD

Cytoplasmic male sterility (CMS) in plant is a phenomenon of fail to produce functional pollen in the course of sexual propagation. It is a frequently encountered phenotype in flowering plants. In 1921,CMS was first found in flax, and so far, it has been found in maize, broomcorn, rice, wheat, rape and sunflower, which is more effective while using heterosis.Great success has been achieved in the development of hybrid varieties in several major crops, including rice and maize. Due to lack of genetic male sterile resources or mutants, soybean is one of a few crop species, in which heterosis has not been exploited to a large extent that is practically viable until recently. The first soybean CMS line was found by Huan Sun et al. in 1993 and great progress of studying and using was made in this field. However, little has been known about its molecular mechanism until now.Twenty one pairs of cytoplasmic male sterile lines and their maintainer lines(JLCMS1-1A,JLCMS1-1B,JLCMS1-2A,JLCMS1-2B......JLCMS1-21A,JLCMS1-21B)were used in this experiment. The mitochondrial DNA from soybean by Randomly Amplified Polymorphic DNA (RAPD) technology was studied, and mtDNA fingerprinting of sterile lines and their maintainer lines were established. This is the foundation for isolation and identification of the relevant genes of CMS soybean, and offering the theoretical foundation for the molecular mechanism of announcing the soybean cytoplasm male sterility (CMS ).The main results are as follows:1. Via repeat experiments the method was fished out for the purification of soybean mitochondrial DNA which is so pure that can be used for the analysis of RAPD.2. A steady and high-efficient reaction system of RAPD has been set up in this study. The optimum conditions in 25μl volume are as follows: 0.1mmol/L dNTPs, 2U Taq DNA enzyme, 0.4ng/μl sample DNA, 0.8ng/μl primer.3. It was proved that we could measure polymorphism of mitochondrial DNA of sterile line and its maintainer by RAPD. Some differential amplified productions of mitochondrial DNA of soybean between sterile lines and their maintainer lines were found. Three...