Mitochondria DNA Polymorphism And Its Relationship With Cold Hardiness In Peach[Prunus Persica(L.)Batsch]

Through the simple extraction of peach mitochondrial DNA(mtDNA), bioinformatics analysis and and PCR, Mitochondria DNA Polymorphism and Relationship with Cold Hardiness in Peach were studyed mitochondria DNA polymorphism and relationship with cold hardiness in peach(Prunus persica(L.)Batsch). It is very important to find molecular marker linked to the tree cold hardiness in the cytoplasm(mitochondrial). It is very helpful to select breeding of peach tree and identify and improve the accuracy of the early selection of parents. There were some chief process and conclusion as follows:This study investigated a reproducible, simple and convenient mitochondrial DNA isolation method via differential centrifugation, sucrose density gradient centrifugation and mitochondria intravital staining. The result indicated that after the high speed tissue machine, high purity mitochondria could be exacted. The process was preliminarily obtained, which can satisfy normal experiment requirements. In addition, the experiment were supply the procedures of peach mitochondrial DNA isolation, laying a foundation for molecular biology research of extranuclear inheritance of mitochondrial DNA.The experimental materials were those peach leaves which were graded by the natural selection of the winter of 2009. 21 varieties included Huichun, Guizhou, GF677, Quzhi, Ruiguang27, Zaoxialu, Fenghuayulu, Zhonghuashoutao, Yangquanroutao, Jilin8701, Jilin8801, Mafim, Shouhong, Honggang, Vallegrande, DesertRed, Wuyuehuo, Zhongning3, Shenzhoubaimi, Huangjinpantao, Qingsi. The tie of specific fragments polymorphism and varieties hardiness of partial sequences of atp9, atp4(orf25), coxI, coxIII, 18s-5srrna, ssu, nad4 exon1/2 were studied via PCR and direct sequencing. The results indicated that we first get partial gene sequences even the amino acid sequence of mitochondrial DNA atp9, atp4(orf25), coxI, coxIII, 18s-5srrna, nad4 exon1/2 were obtained. There was no difference among 21 varieties expect partial sequences of atp9.G base in-96 bp ~-94 bp of open reading frame upstream noncoding region deletion or not made a distinction between cold resistance and tender varieties, the accuracy rates was 81%. Because of the structure of atp9 was few studied, there may be the circuit changer in expression regulation or RNA structure.