Abstract: A fast and high productivity method for mitochondrial DNA(mtDNA)isolation in Chinese cabbage was introduced.Yellow seedlings were used in order to remov chloroplast DNA(cpDNA).DNase I and RNase A enzyme were added to digest nuclear DNA(nDNA)and RNA in order to purify mitochondrion.Protein K and SDS were added to disrupt mitochondrial membranes and 2 steps of poeeilothermal treatments with 50.C for 1 h and 37℃for 1 h were applied to release more mtDNA.Results showed electrophoretic band of mtDNA was clear and the purity Was high.The ratio of OD2∞/OD2帅was about 1.8.No products were amplified for mtDNA template by using primers deigned by anclear housekeeping Actin genes.It meant mtDNA Was not contaminated by nDNA.670 bp fragment could only be amplified in CMS lines by using primers deigned by orf222 gene special for Chinese cabbage CMS96.RAPD analysis with 5 primers showed many polymorphic bands were founded between CMS96 and maintainer lines.Therefore mtDNA isolated in this method is suitable for molecular manipulation.The innovation in this method is that 2 steps of poecilothermal treatments with 50℃for 1 h and 37℃for 1 h are used to disrupt mitochondrial membranes and make higher productivity of mtDNA.The highest yield in our study is 18.83¨g mtDNA for 1 gram yellow seedlings.

Key words: Chinese cabbage, M itcoehondrial DNA, Isolation, Poecilothermal treatoents, CMS

CLC Number: 

• S634.1