改进的CTAB法提取32种中药破壁饮片DNA及物种鉴定

Extraction of DNA from 32 Kinds of Chinese Herbal Pieces by Improved CTAB Method and Species Identification

中药破壁饮片是由传统中药饮片经过气流破壁技术加工而成,已失去了原药材原有的外观性状特征,传统的鉴别方法已很难鉴别其真伪。本研究选用ITS2序列作为DNA条形码对32个中药破壁饮片品种进行研究,验证DNA条形码技术应用于中药破壁饮片上的可行性。通过提取32个品种的DNA、PCR扩增及双向测序获得ITS2序列,在中药材DNA条形码鉴定系统和NCBI数据库中进行比对。实验结果表明:32个品种通过改良的CTAB法均能成功提取DNA,PCR扩增及测序后均能得到高质量的ITS2序列。因此,基于ITS2序列的DNA条形码技术可有效地鉴定中药破壁饮片,为中药破壁饮片真伪鉴别提供有效手段。

Ultrafine granular powder of Chinese medicine is processed by the traditional Chinese medicine decoction pieces through airflow breaking technology.It has lost the original appearance characteristics of the original medicinal materials,and traditional identification methods have been unable to identify its authenticity.We choose 32 varieties in this study,ITS2 sequences were selected as DNA barcodes to study all varieties of ultrafine granular powder of Chinese medicine,to verify the feasibility of applying DNA barcode technology to ultrafine granular powder of Chinese medicine.ITS2 sequence was obtained by extraction of 32 strains of DNA,PCR amplification and bidirectional sequencing.BLAST was performed in the barcode identification system and Pubmed database of Chinese herbal medicines.The result shows that DNA of 32 samples can be successfully extracted by modified CTAB method,and high quality ITS2 sequences can be obtained after PCR amplification and sequencing.Therefore,based on the ITS2 sequence of DNA barcode technology can effectively identify the ultrafine granular powder of Chinese medicine,to provide a useful method for the identification of ultrafine granular powder of Chinese Medicine.