Complete Sequence Analysis Of Mitochondrial DNA In Pinus Taeda

In this study,the loblolly pine clone,202,was selected as the research material,and five methods of DNA extractions were screened for better DNA quality.The high quality loblolly pine DNA was sequenced through the Solexa high throughput sequencing principle,and the mitochondrial DNA(mitochondrial deoxyribonucleic acid,mt DNA)was enriched,then the obtained mt DNA sequence was analyzed,and the gene functional annotation was arranged.The main results were as follows:(1)Five kinds of DNA extraction methods were: 3 traditional extraction methods(CTAB,SDS,Zigenhagen)and 2 extraction kits(Tian Gen extraction kit,Sheng Gong extraction Kit).The concentration of DNA obtained by the traditional CTAB method was the highest,and there was a significant difference between the concentration of DNA extracted from the other four methods(P<0.05),which was in accordance with the results of gel imaging.The OD260/OD280 of DNA proposed by CTAB and Tian Gen kit were slightly more than 2,and there was a small amount of RNA contamination.The OD260/OD280 of SDS and Sheng Gong kit was slightly less than 1.8,indicating a small amount of protein and other macromolecules.The purity of DNA extracted by Zigenhagen method was the best,and the OD260/OD280 value was 1.8-2.0,and there was no impurity pollution.To sum up,in the extraction method of needles DNA of Pinus taeda,CTAB method and Tian Gen kit method were better than the other methods.(2)The DNA of loblolly pine was sequenced by Solexa high throughput sequencing.The length of the mitochondrial genome was 1,191,054 bp,the proportion of GC was47.77%,with no gap,and the sequencing data was complete.(3)There were 84 genes in the mitochondrial DNA of Pinus taeda,including 11 types,which were 7 genes encoding the r RNA,3 genes encoding the cytochrome C oxidase subunit,14 genes encoding the ribosomal protein,9 genes encoding the NADH dehydrogenase subunit,38 t RNA-amino acid genes,and 5 genes encoding the ATPase subunit,4 genes encoding cytochrome c biogenesis,1 gene encoding Sec-independent protein translocase protein,1 gene encoding maturase,1 gene encoding succinate dehydrogenase cytochrome subunit and 1 encoding apo cytochrome b.