Optimization of DNA extraction methods and molecular identification of endophytic Xylaria from Dendrobium
This study aims at optimizing DNA extraction methods of endophytic Xylaria from Dendrobium and authenticating the fungus based on ITS and secondary structures of ITS2.Extraction effects of kit method and CTAB method at different concentration (2％,3％,4％) were compared and CTAB method was optimized.ITS sequence was amplified and the secondary structure of ITS2 was predicted.Results indicated that the best extraction effect was derived from CTAB at concentration of 4％ and the optimized CTAB method was greatly reduced DNA extraction duration.The ITS sequence of experimental Xylaria strain TJ1 showed 99％ similarity to that of three species of Xylaria,X.arbuscula,X.apiculata and X.venosula,while the secondary structure of ITS2 of the strain showed 100％ similarity to that of Xylaria arbuscula.The optimized CTAB method will provide convenience of future research and the endophytic Xylaria of Dendrobium can be identified accurately based on combining ITS sequence with secondary structures of ITS2.
NETL
NSTL
万方数据
维普
