银杏内生小孢子链格孢的分离与鉴定

Isolation and identification of endophytic small-spored Alternariain Ginkgo biloba L.

【目的】对银杏内生小孢子链格孢进行分类鉴定,探索鉴别内生小孢子链格孢的有效方法。【方法】采用组织分离法分离银杏叶片中的小孢子链格孢,分别用产孢表型鉴定法和OPA2-1核苷酸序列分析法对分离的小孢子链格孢进行初步鉴定。【结果】从银杏叶片中分离到55株内生小孢子链格孢,老叶中内生小孢子链格孢的分离几率(7.8%)明显比新叶(4.3%)高,是新叶的1.8倍。根据产孢表型可将55个分离物归为5个类群;通过OPA2-1核苷酸序列及系统发育分析,将银杏内生小孢子链格孢分为2个聚类组,其中分离物N02-4和XL-7聚在一个独立的分支上,而其余9个分离物与细链格孢(A.alternaria)、细极链格孢(A.tenuissima)、粗柠檬链格孢(A.limoniasperae)聚为一类,未能反映遗传上的差异。分离物N02-4的产孢表型与其他分离物不同,且其OPA2-1核苷酸序列与其他分离物有3个碱基的差异;分离物XL-7的产孢表型与细极链格孢(A.tenuissima)一致,但在核苷酸序列上存在3个碱基的差异。【结论】仅根据产孢表型对银杏内生小孢子链格孢进行归类,信息有限,结果不准确,需结合分子生物学手段才能进行准确归类。

【Objective】Endophytic small-spored Alternariain Ginkgo biloba L.were classified and i-dentified to explore effective approach to identify small-spored Alternaria.【Method】Small-spored Alternaria were isolated fromGingko biloba L.leaf tissue.Small-spored Alternaria were identified and analyzed by sporulation phenotype and OPA2-1 nucleotide sequences respectively.【Result】Total 55 isolates of endophytic small-spored Alternaria were isolated fromGinkgo biloba L.leaves.Isolation rate of endophytic small-spored Al...