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        藥用植物羌活種子DNA條形碼鑒定研究

        發(fā)布時(shí)間:2019-08-26 來(lái)源: 歷史回眸 點(diǎn)擊:

        http://img1.qikan.com.cn/qkimages/zgyi/zgyi201603/zgyi20160305-1-l.jpghttp://img1.qikan.com.cn/qkimages/zgyi/zgyi201603/zgyi20160305-2-l.jpghttp://img1.qikan.com.cn/qkimages/zgyi/zgyi201603/zgyi20160305-3-l.jpg
          [摘要]基于ITS2 條形碼對(duì)藥用植物羌活種子進(jìn)行分子鑒定,以確;_。該研究對(duì)藥用植物羌活種子進(jìn)行形態(tài)特征比較,每份樣品進(jìn)行3次隨機(jī)取樣,對(duì)每次取樣的單粒種子提取基因組DNA,PCR擴(kuò)增,雙向測(cè)序并拼接獲得ITS2序列。通過(guò)遺傳距離法、建樹(shù)法,并結(jié)合中藥材DNA條形碼鑒定系統(tǒng)()進(jìn)行基原鑒定。結(jié)果顯示,羌活與寬葉羌活種子形態(tài)特征相似,難以明顯區(qū)分。經(jīng)分子鑒定,27份種子樣品有24份樣品為2010年版《中國(guó)藥典》規(guī)定的羌活正品基原種子,包括13份羌活種子,11份寬葉羌活種子,另外3份樣品不屬于羌活基原植物種子。研究證明,DNA條形碼技術(shù)可以準(zhǔn)確鑒別羌活種子基原,為中藥材種質(zhì)資源鑒定提供了新方法,對(duì)中藥材種子質(zhì)量標(biāo)準(zhǔn)的建立提供依據(jù)。
          [關(guān)鍵詞]中藥材種子;羌活;ITS2; 分子鑒定
          [Abstract]In order to guarantee the species correction of Notopterygium seeds, a molecular identification method with ITS2 as DNA barcode has been verified. In this study, 27 samples of Notopterygium seeds were collected from the main producing area of Notopterygium. The morphological characteristics of the Notopterygium seeds were firstly surveyed. Then the DNA extraction, PCR amplification, DNA sequencing and DNA assembly were carried out. The species identification for a Notopterygium seed was implemented through distance method, NJ-tree method and the DNA barcoding system for traditional Chinese medicine (). The results showed that the seeds of N. incisum and N. franchetii had similar morphological characteristics and were difficult to distinguish clearly based on morphological descriptions. With the results of molecular identification, 24 samples were genuine including 13 N. incisum seeds samples and 11 N. franchetii genuine seeds samples. In conclusion, DNA barcode technology can accurately and efficiently identify the species of Notopterygium seeds. Furthermore, this study will provide a new method for germplasm resources identification of medicinal materials and supplies some guidelines for establishing Chinese herbal seeds and seedlings quality standards.
          [Key words]Chinese herbal seeds; Notopterygium; ITS2; molecular identification
          中藥材種子是中藥材生產(chǎn)、流通、使用的源頭,種質(zhì)資源的優(yōu)劣直接影響中藥材的產(chǎn)量與質(zhì)量,準(zhǔn)確鑒定中藥材種子,確保種質(zhì)來(lái)源純正可靠,既為大規(guī)模規(guī)范種植生產(chǎn)中藥材打下基礎(chǔ),也為中藥材的臨床用藥安全提供保障。本研究以中藥材羌活種子為研究對(duì)象,首次將DNA條形碼技術(shù)由中藥材鑒定領(lǐng)域引入到中藥材種子鑒定中,為后續(xù)中藥材種子鑒定研究提供依據(jù),對(duì)中藥材種子質(zhì)量標(biāo)準(zhǔn)的建立有重要意義。
          中藥材羌活為傘形科植物羌活Notopterygium incisum Ting ex H.T.Chang或?qū)捜~羌活N. franchetii H.de Boiss.的干燥根莖和根[1],為傳統(tǒng)中藥成方藥材,用藥需求量大。羌活植物生長(zhǎng)周期長(zhǎng),受高海拔生長(zhǎng)環(huán)境和自身發(fā)育特性的影響,羌活種子成熟度不夠,發(fā)芽率較低,使羌活資源更替困難,加上過(guò)度采挖,野生羌活資源日益減少[2]。羌活被列入國(guó)家重點(diǎn)保護(hù)野生藥材物種名錄,為42種國(guó)家重點(diǎn)保護(hù)的野生動(dòng)植物藥材品種之一。規(guī);斯しN植繁育是解決羌活資源危機(jī)的主要途徑,有關(guān)羌活種質(zhì)特性和發(fā)芽率的研究報(bào)道不斷增多且取得一定成效[3-5],這對(duì)中藥材羌活資源庫(kù)的恢復(fù)重建有重要意義。本研究基于DNA條形碼技術(shù)對(duì)羌活種子樣品鑒定,以保證中藥材羌活種子基原真實(shí)準(zhǔn)確,將極大促進(jìn)羌活種質(zhì)資源的保護(hù)開(kāi)發(fā)。
          1材料
          從甘肅、四川、青海等中藥材羌活主產(chǎn)區(qū)購(gòu)買(mǎi)27份藥用植物羌活種子樣品,實(shí)驗(yàn)樣品信息見(jiàn)表1。
          2方法
          2.1形態(tài)學(xué)考察樣品為羌活種子,于LY-WN-HPCCD型動(dòng)態(tài)顯微成像系統(tǒng)(成都勵(lì)揚(yáng)精密機(jī)電有限公司)下觀察種子外部形態(tài)(形狀、大小、顏色,表面紋理),拍照并記錄,游標(biāo)卡尺(上海九量五金工具有限公司)精確測(cè)量長(zhǎng)、寬、厚及延展翅長(zhǎng)度,并用百粒法測(cè)其千粒重[6],確保取樣均勻性和代表性。

        相關(guān)熱詞搜索:羌活 條形碼 鑒定 種子 藥用植物

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