| 摘要: |
| 【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。 |
| 关键词: 银杏 内生小孢子链格孢 产孢表型鉴定法 OPA2-1核苷酸序列分析法 |
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| 基金项目:国家科技支撑计划项目（2007BAI32B04)；西藏自治区地区基金项目（2007-自然基金-25)；国家自然科学基金项目(30760005) |
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| Isolation and identification of endophytic small-spored Alternaria in Ginkgo biloba L. |
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| Abstract: |
| 【Objective】Endophytic small-spored Alternaria in Ginkgo biloba L.were classified and identified to explore effective approach to identify small-spored Alternaria.【Method】Small spored Alternaria were isolated from Gingko 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 from Ginkgo biloba L. leaves.Isolation rate of endophytic small-spored Alternaria from old leaves（7.8%） was higher than that of young ones（4.3%）,1.8 times of the young leaves.The 55 isolates were categorized into 5 groups based on sporulation patterns,but grouped into 2 genotypes by nucleotide sequences of OPA2-1.Isolates N02-4 and XL-7 could be differentiated from other isolates,which gathered in the same group,other isolates gathered in the same group with A.alternaria,A.tenuissima and A.limoniasperae.Sporulation phenotype of isolate N02-4 was different from other isolates,and its OPA2-1 nucleotide sequence with other isolates had three basic differences.Isolate XL-7 was the same on sporulation patterns but 3 different bases on nucleotide sequences of OPA2-1 with A.tenuissima.【Conclusion】By comparing two methods,sporulation patterns could only give us limited information with inaccurate result,and can be accurately classified by combination of molecular biology means. |
| Key words: Ginkgo biloba L. endophytic small-spored Alternaria sporulation phenotypic identification method OPA2-1 nucleotide sequence analysis |
