【摘要】：利用ISSR分子技術(shù),篩選引物用于構(gòu)建灰葡萄孢(Botrytis cinerea)不同發(fā)育表型群體的指紋圖譜。從收集到的300株灰霉菌中選35株代表菌株,雖然其菌落形態(tài)有所差異,但經(jīng)形態(tài)學鑒定、ITS鑒定、特異引物BC729鑒定及三基因(G3PDH、HSP60和RPB2)系統(tǒng)發(fā)育樹分析均表明其為灰葡萄孢種群。在此前提下,篩選可用于區(qū)分不同發(fā)育表型的ISSR引物,引物所擴增的圖譜要求穩(wěn)定重復(fù),主帶明顯,帶間距寬。經(jīng)篩選獲得3條引物,名稱分別為807、864和886,其最優(yōu)退火溫度分別為51℃、52℃和49℃。供試35個代表菌株經(jīng)引物807擴增后出現(xiàn)3種不同類型指紋圖譜,經(jīng)引物864擴增后出現(xiàn)4種不同類型圖譜,經(jīng)引物886擴增后出現(xiàn)5種不同類型圖譜。將以上3個引物的擴增圖譜聯(lián)合分析,本研究供試菌共出現(xiàn)9種圖譜組合。9個圖譜類型對應(yīng)不同的菌落形態(tài)�；移咸焰叻N下存在高水平分化現(xiàn)象,由于其菌落形態(tài)受培養(yǎng)環(huán)境影響較大,僅憑形態(tài)區(qū)分容易造成誤差,本研究搭建了一個灰葡萄孢種下發(fā)育表型的分子鑒定平臺,為今后開展菌群鑒定與分析、不同研究條件下菌群比較、演化更替及發(fā)掘新菌群提供研究手段。
[Abstract]:ISSR molecular technique was used to screen primers to construct fingerprints of different phenotypic populations of (Botrytis cinerea). 35 representative strains were collected from 300 strains of Ash fungus. Although their colony morphology was different, they were identified as grapevine species by morphological identification, specific primer BC729 identification and phylogenetic tree analysis of three genes (G3PDHU HSP60 and RPB2). On this premise, ISSR primers can be used to distinguish different developmental phenotypes. The amplified map of the primers should be stable and repetitive, the main band is obvious, and the band spacing is wide. Three primers, named 807864 and 886, were obtained. The optimum annealing temperatures were 51 鈩，

本文編號：2244857