DNA條形碼技術(shù)在常見中藥材蛇類鑒別中的應(yīng)用

　　[摘要] 中藥材的準(zhǔn)確鑒別是中藥材研究、生產(chǎn)和應(yīng)用至關(guān)重要的一步。DNA 條形碼技術(shù)以線粒體細(xì)胞色素氧化酶Ⅰ亞基（COⅠ）基因序列作為標(biāo)記，在中藥材鑒別中的應(yīng)用日益增多。研究應(yīng)用DNA 條形碼通用引物擴(kuò)增測序，探討DNA條形碼技術(shù)在常見中藥材蛇類（6科15屬19種共109個樣品）鑒別的可行性，采用鄰接法構(gòu)建該類群分子系統(tǒng)發(fā)育樹。結(jié)果表明，該片段的G+C平均量為43.9%，低于A+T平均量（56.1%）�；�于Kimura 雙參數(shù)模型計算，白唇竹葉青、烏梢蛇和赤練蛇的種內(nèi)平均遺傳距離均大于2%。通過構(gòu)建系統(tǒng)發(fā)育關(guān)系后進(jìn)一步分析發(fā)現(xiàn)，白唇竹葉青一樣品鑒別有誤。烏梢蛇中的某些樣品也可能存在鑒別有誤，即原本是灰鼠蛇的可能被錯誤鑒定為烏梢蛇。造成赤練蛇種內(nèi)遺傳距離差異大的因素需進(jìn)一步分析。DNA條形碼用于常見中藥材蛇類種間的鑒別是可行的，極大程度地彌補(bǔ)統(tǒng)形態(tài)學(xué)分類方法的缺陷，值得加大推廣應(yīng)用和進(jìn)一步深入研究。
　　[關(guān)鍵詞] COⅠ;遺傳距離;系統(tǒng)分類;烏梢蛇
　　[收稿日期] 2014-08-28
　　[基金項目] 國家自然科學(xué)基金項目（31460559）;廣西自然科學(xué)基金項目（2014GXNSFBA118113）;廣西科技廳項目（桂科重12118005-2-5）;南寧市科技攻關(guān)項目（20123238）;廣西分析測試研究中心基本業(yè)務(wù)費項目（2012ACZ07）
　　[通信作者] 李力，E-mail： liboshi1963@vip.163.com;林葵，E-mail： linkui8910@163.com
　　[作者簡介] 黃勇，博士，主要從事藥用動物系統(tǒng)分類和分子進(jìn)化研究，E-mail： huangykiz@163.com
　　DNA barcoding and its utility in commonly-used medicinal snakes
　　HUANG Yong， ZHANG Yue-yun， ZHAO Cheng-jian， XU Yong-li， GU Ying-le， HUANG Wen-qi， LIN Kui， LI Li
　�。�1. Guangxi Botanical Garden of Medicinal Plants， Nanning 530023， China;
　　2. Guangxi Research Center of Analysis， Nanning 530022， China）
　　[Abstract] Identification accuracy of traditional Chinese medicine is crucial for the traditional Chinese medicine research， production and application. DNA barcoding based on the mitochondrial gene coding for cytochrome c oxidase subunitⅠ（COⅠ）， are more and more used for identification of traditional Chinese medicine. Using universal barcoding primers to sequence， we discussed the feasibility of DNA barcoding method for identification commonly-used medicinal snakes （a total of 109 samples belonging to 19 species 15 genera 6 families）. The phylogenetic trees using Neighbor-joining were constructed. The results indicated that the mean content of G+C（46.5%） was lower than that of A+T（53.5%）. As calculated by Kimera-2-parameter model， the mean intraspecies genetic distance of Trimeresurus albolabris， Ptyas dhumnades and Lycodon rufozonatus was greater than 2%. Further phylogenetic relationship results suggested that identification of one sample of T. albolabris was erroneous. The identification of some samples of P. dhumnades was also not correct， namely originally P. korros was identified as P. dhumnades. Factors influence on intraspecific genetic distance difference of L. rufozonatus need to be studied further. Therefore， DNA barcoding for identification of medicinal snakes is feasible， and greatly complements the morphological classification method. It is necessary to further study in identification of traditional Chinese medicine.

相關(guān)熱詞搜索：蛇類 條形碼 中藥材 鑒別 常見