The Technology Of New Pattern Molecular Markers And Genetic Markers On Rice Male Sterility

Rice is the world's extensive cultivation of important grain crops. It has made great contributions that the success of hybrid rice breeding in rice cultivation is a major breakthrough for the development of grain production. Male sterility is one of the main way in the use of rice heterosis. Molecular marker technique can accurate to expression of species of genetic information which is the new markers depond on DNA form to manifest genetic character.There are many of PCR-based molecular markers. But their development and application are limited by theirself shortcomings. AFLP is widly applicated for many merits. It is high reliability and high efficiency, and has the codominance and recessiveness, the highest level of polymorphism, the most testing sites, the highest rate of molecular recognition, faster, less DNA needed and so on. But it requests highly technical, and must carry on the enzyme to cut with the connector lug. This study does PCR by designed primers which with restriction site sequence, to expect to establish a new tag technique similar AFLP but without enzyme cut DNA and link.Based on restriction enzyme PstI identify the DNA sequence, the study designs a series of simple primers (5' -NNNMCTGCAGMNN) to PCR amplification, then to the various mark technology parameter.Using two selected new primers which are bands clear and stability good, the study carries on PCR analysis to 13 varieties of rice genomic DNA, and the optimization of PCR reacting system and the different electrophorsis method detected. On this basis, the study did PCR comparative analysise the resluts of the same primer on different DNA samples, and different primer for the same DNA samples, the new primer and AFLP primer PCR amplification, and the quality of rice for each other male non-nuclear fertility on the basis of genetic, new primer on male sterility rice conducted a preliminary study on the tags. This study shows that:1,The new primer is proved stability, reliability and a certain polymorphism, from the comparative analysis the new primer on the different DNA samples of rice, different crops; different new tag primer on the same template; new tag primer AFLP primer amplification.2,Compare with the agarose and the polyacrylamide two detection methods, the certificate new marker primer more suitable for denaturing polyacrylamide gelation methodcarries on the examination.3,Fertility of rice genetic analysis shows that: Jin A,Te A,You IA was sterility of the two major gene control.4,Teyou 253 F₂ CMS of individual identification, the establishment of a "fertile pool" and "sterile pool," and with new primer Pp31-42, Pp43-32 for a tag, failed to detect differences in point.