RAPD Analysis On MtDNA Of The Three Alloplasmic CMS With Isonuclear And Its Cloning And Sequence Of Special Fragment

Cytoplasmic male sterility (CMS) is ubiquitous in nature as a maternal plant genetic phenomenon and which is an important basis in Crop Heterosis Utilization at the same time. Wheat as the main food crop, studying its utilization of heterosis and male sterility is the matter that many breeders have been trying to overcome. Many studies have shown that there are some direct relationships between mitochondrion and CMS in wheat, Exploration of the relationship between mitochondrial DNA (mtDNA) and CMS is very important in studyinging the mechanism of CMS in wheat at present.In this study, male sterile lines in wheat with the same male parent nucleus of 90-110, cytoplasm from kotschyi,bicornis and the T.spelta were used as materials.There was a depth study on the ms (Ae.kotschyi)-90-110,ms (Ae.bicornis)-90-110 and ms(T.spelta)-90-110 and obtained some important results.the main results were as follows:1:Special fragment were amplified using three primers of 50 primers.OPY01-1517 was a special fragment amplified using the primer of OPY01 in ms(Ae.bicornis)-90-110. OPA05-2500 was amplified using the primer of OPA05 in(T.spelta)-90-110. OPP02-750 was amplified using the primer ofof OPP02 in(Ae.kotschyi)-90-110.2:The three above-mentioned special fragments are stable and highly repeatable. The results showed that there was an apparent polymorphism in the three case of male sterility of wheat,and this polymorphism is likely to be the main reason of the difference of restoration and fertility in three case of wheat. At the same time, these markers can be used as marker genes related to male sterility.3:OPY01-1517,OPA05-2500 and OPP02-750 were successfully cloned and sequenced. The three sequences were used to blast the Genbank databases. The results showed that OPY01-1517 and OPP02-750 sequence had 99%and98%similarity with some triticum aestivum mitochondrial genome sequences (wheat mtDNA Genome accession number EU534409.1). OPY01-1517 contained atpl gene sequence and the sequences is located at 3' end, the length is 106bp. The remaining sequence of non-coding sequence. OPP02-750 is the non-coding sequence of triticum aestivum mitochondrial genome of nadl and nad5. These two sequences can be use to distinguish the two wheat varieties of ms(T.spelta)-90-110 and ms(Ae. bicornis)-90-110.