張引芳; 法朗西斯·莫林娜
上海市農(nóng)科院食用菌研究所; 美國菌種保藏中心 上海 201106

【中文摘要】 本文應(yīng)用RFLP技術(shù)鑒別兩未知菌株S1001和S1002是否系鱗翅目蛹蟲草菌(Cordyceps milicaris)。試驗以ITSl,TTS4為引物,PCR擴(kuò)增rDNA的5.8S+ITS區(qū)段,擴(kuò)增產(chǎn)物純化后用DNA限制性內(nèi)切酶酶切,獲得DNA的RFLP圖譜,并同ATCC模式菌株的RFLP圖譜相比較。結(jié)果表明:三個作為對照的ATC菌株具相同的RFLP圖譜,而兩待鑒定菌株之間存在差異,且明顯不同于ATCC模式菌株,從而證明,兩待鑒菌株與ATCC菌株之間在rDNA區(qū)域具有明顯的結(jié)構(gòu)差異,故推斷兩待鑒定菌株不隸屬于鱗翅目蛹蟲草菌。

【英文摘要】 RFLP was applied in order to identify if two unknown cultures--S1001,S1002were correctly assigned to Cordyceps militaris. Restriction analysis of enzymatically amplified ribosomal DNA was used to compare fingerprints of these cultures with three ATCC "reference" strains of C. militaris. DNA was extracted from cultures grown in liquid medium and the region spanning of the 5.8S rRNA gene and the two internal transcribed spaces (5.8S + ITS) were amplified with the Polymerase Chain Reaction (PCR). The PCR products were purified and then subjected to double digestion with three pairs of cndonucleases. For all enzyme pairs, the three ATCC strains exhibited identical profiles that were easily distinguishable from those of the two unknown strains, which showed very different fingerprints. The results suggested that the two cult-arts were incorrectly assigned to C. militaris.

【中文關(guān)鍵詞】 PCR; RFLP; 要; 菌株鑒別; 鱗翅目蛹蟲草菌
【英文關(guān)鍵詞】 PCR; RFLP; Identification; Cordyceps militaris

【文獻(xiàn)出處】 食用菌學(xué)報,Acta Edulis Fungi,編輯部郵箱,1994年02期 【DOI】CNKI:SUN:SYJB.0.1994-02-005