Abstract: Using etiolated seedlings of 2 sesame varieties,an economic,rapid,efficient sesame seed mitochondria DNA(mtDNA) extraction method was established through the key factor DNase digestion time,including homogenizing processing,differential centrifugation comprising 2 400 g centrifugation for 15 min and 12 000 g centrifugation for 12 min to acquire primary mitochondria,DNase digestion for 1.5 h to eliminate the pollution of genome DNA,centrifugation with 0.6 mol/L sucrose padding to get pure mitochondria,and SDS-proteinase K lysis to obtain pure mtDNA.The mtDNA samples were tested using DNA/Protein Analyzer,PCR with genome-special markers(nuclear gene-specific primer beta-actin,mitochondrial gene-specific primer rrn26S and chloroplast gene-specific primer ARCP5) and 1.0%agarose gel electrophoresis.It was proved that the mtDNA isolated by this method not only has the yield more than 1 μg per gram material,but also characterized of high purity.The results showed that this high quality mtDNA can be successfully used in genetic studies as sesame cytoplasmic male sterility and stress resistance.

Key words: Sesame, mtDNA, Extraction, DNase treatment

CLC Number: 

• S6.03