Soybean Restorer Fertility Related Genes Aflp Differences Fragment Screening, Cloning, And Scar Conversion

Soybean is a typical plant of self-pollinate, and it is an important resourse of seeds. As one of the major crops in China and all over the world, soybean plays very important role in world food supply and trade market. Utilization of heterosis is the important and useful way to promote crop output and quality. One of the obstacle to utilize soybean heterosis is that most of the alien cytoplasm has harmful effect and has no excellent restoreing. Male sterility with cytoplasm has been proved to be promising in hybrid soybean breeding, which has broad spectacle of restoring gene resources and has no distinct harmful cytoplasmic effect. While there are less restoring lines with both higher restoring ability and excellent agronomic performance in soybean, so it is impossible to use them directly in hybrid production. Breeding of restorer is one of the prerequisite to produce hybrids using CMS system, and is more difficult than conventional breeding, because the presence of restoring genes in breeding lines cannot be detected through traditional approaches without a generation of progeny testing, which is lab and time-consuming to cross every plant of a segregating population with male sterile line, so the plant with excellent agronomic traits often has no Rf gene and the restoring ability gets stable slowly through generations.The utilization of DNA molecular markers will be beneficial to overcome the difficulty of restorer breeding. With molecular markers, it is possible to detect segregating plants during seedling stage without plenty of test crossing and examining work, make the trait of restoring ability get stable quickly, raise accuracy in selecting restorations, pyramid minor RF genes, raise restoring ability and apply it to the management of hybrid seeds. At the same time, we may locate Rf genes precisely by screening molecular makers linked to them, and this may provide a basis for isolating Rf genes.In this research, soybean is used as a representative genic male sterile AB lines, and it is also an optimal plant material in the research of male sterility mechanism. In order to compare the differences of genomic DNA between the restoring line and the sterile line with AFLP technique, genomic DNA bulks of the restoring and the sterile line of which were built up. By using different primer combinations, some polymorphism bands were determined. Meanwhile, the cloning and sequencing of which was also performed and the main results were as follows:1) Only one obvious polymorphism band was obtained in the restoring line from selective PCR amplification with 64 pairs E+3/M+3 primer combinations, and which was named as ME10-3.2) It proved that there should be nucleotides sequence similar to ME10-3 in the sterility line by the Southern hybridization using the fragment ME10-3 as probe. Basing that, the mutation of nucleotides sequence in the region related to...