Research And Application Of Sinomenium Acutum DNA Barcoding

DNA barcoding technology is the latest development in the field of biological molecular identification, that by using a standard, enough variation, short and easy to enlarge DNA fragments in the genome to identify species. This identification technology is easy to master, can identify the specificity of the intraspecific and interspecific diversity, while is conducive to the protection of the rare species, and find new species. This study mainly discusses the analysis of DNA barcoding technology used in Sinomenium acutum medicinal plants, to identify Sinomenium acutum and its adulterant species, establish Sinomenium acutum DNA barcode development process, for in the market circulation of Sinomenium acutum a radical effect of herbs.The main results are as follows:1.By comparing the CTAB method, high salt and low pH method and two kinds of kits for extraction method of DNA,best extraction method of DNA from TIANGEN plant genome extraction kits was found,on the basis of this extraction method of DNA of Chinese herbs was optimized;In this study universal plant primer of DNA barcode was used,according to the description of literature on these universal plant primer PCR conditions and procedures,the annealing temperature and cycle times of PCR reaction were adjusted to facilitate the study.2. Sinomenium acutum 4 candidate DNA barcodes (ITS2, psbA-trnH, matK, rbcL) were amplified and sequenced,the data obtained was processed and analyzed by bioinformatics software,according to the principle of selection for DNA barcoding,comparing the distance of inter/intra species between the 4 fragments,ITS2 was the most suitable fragment for Sinomenium acutum DNA barcode.3.Using DNA barcoding technology based on ITS2 sequences 10 samples from different areas of herbs were identified,identified successfully of Sinomenium acutum genuine herbs and its adulterants Sabia swinhoei and Paederia scandens.Although the processing or processing of some medicinal herbs has an impact on molecular identification, the DNA bar code technology can solve most of the problems that have been identified in the authenticity of Chinese herbal medicines.