Preliminary Study On Transferring Of Mitochondria And Its MtDNA In Wheat

It is always comprehensive regarded to utilize wheat hybrid vigor after wheat male sterile was founded. It has very important practiced significance to improve more yield and quality of wheat. Male sterile is the base of the utilization of crop hybrid vigor. This research does the attempt of transferring of wheat mitochondria and mitochondrial DNA. It is hoped to change existed cultivation pattern of male sterile, establish the method of directional creating male sterile, and conquer the bad effect of the cytoplasm. And especially it does the theory work of the relationship between wheat CMS and mitochondria. The materials of this research are wheat sterile line and maintainer line of S-type, K-type, Ven-type and T-type. The results are as follows: 1. The study establishes a quickly method of extracting high actively mitochondria from yellow seeding of wheat depending on key technique of ferential centrifugation and density gradient centrifugation (isodensity centrifugation). And it is compared the activity, purity, and the ultrastructure of the mitochondria extracted by ferential centrifugation, density gradient centrifugation and the improved the density gradient. 2. On the basic of the quickly exacted high active, the study analyzes many facets of the step of exacting wheat mtDNA, and establishes a quickly extracting mtDNA method. 3. RAPD molecular labeling technique is used, which is analyzed the mtDNA polymorphism of the wheat sterile line and maintainer line of S-type, K-type, Ven-type and T-type by primer S22 screen out steady molecular labeling of the mtDNA with these CMS. 4. It is investigated that transfer mitochondria and mitochondrial DNA by the method of stigma dripping and ovary injection. The result indicates that the seed rate of the method of stigma dripping is obviously more than the method of ovary injection. Although there is no fertility transformation in F1, it is obviously changed on the height of the plant, and it is also found the donor strip on analyzed RAPD of the laminae of plant F1. 5. It is tentative investigated the transferring of mitochondria by microinjection. It is found that the key of the success of transferring of mitochondria by microinjection is not the microinjected from pollen grain trema, but the cultivated of the microinjected pollen grain.