人工栽培蛹蟲草退化現(xiàn)象的分子分析
MOLECULAR ANALYSIS OF DEGENERATION OF ARTIFICIAL PLANTED CORDYCEPS MILITARIS
李美娜 吳謝軍 李春燕 馮伯森 李慶偉
摘 要：利用PCR-RFLP和RAPD方法對野生馴化蛹蟲草及其退化菌種進(jìn)行了基因水平的分析.PCR-RFLP實驗,采用真菌通用引物ITS1和ITS4,擴(kuò)增出5.8S和其兩端的兩個轉(zhuǎn)錄間隔區(qū)(ITS),選用5種識別四個堿基的內(nèi)切酶(HaeIII、AfaI、TagI、AluI和XspI),其中只XspI酶切結(jié)果在兩個菌種中存在差異,進(jìn)一步測序表明,整個片段全長534bp,共有13個位點(diǎn)發(fā)生堿基突變,且都為c轉(zhuǎn)換為t.RAPD實驗,選用4組計80個隨機(jī)引物進(jìn)行PCR擴(kuò)增,共篩選出9個對所有供試個體均存在明顯差異的引物.結(jié)果表明了退化菌株較野生馴化菌株在DNA水平上發(fā)生了頻率較高的突變,這些突變可能直接與導(dǎo)致菌種退化的基因相關(guān)聯(lián).
關(guān)鍵詞：馴化菌株, 退化菌株, 5.8SrRNA, RFLP, RAPD
分類號：Q939.53 文獻(xiàn)標(biāo)識碼：A
文章編號：1007-3515(2003)02-0277-0282
基金項目：大連市科技局重點(diǎn)推廣項目:(2002-05)
作者簡介：李慶偉,通訊作者: email; liqw@yahoo.com.cn
作者單位：李美娜（遼寧師范大學(xué)生命科學(xué)學(xué)院,大連,116029）
吳謝軍（大連雙天新藥研究開發(fā)中心,大連,116031）
李春燕（大連雙天新藥研究開發(fā)中心,大連,116031）
馮伯森（遼寧師范大學(xué)生命科學(xué)學(xué)院,大連,116029）
李慶偉（遼寧師范大學(xué)生命科學(xué)學(xué)院,大連,116029）
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