基于RAPD标记的昆虫病原人类绿僵菌的遗传多样性来源于椰子的可耕地粘土

Desianty Dona Normalisa Sirait, M. C. Tobing, Irda Safni
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摘要

金龟子绿僵菌(mettarizhium anisopliae, Metsch.)是一种昆虫病原真菌,主要攻击犀牛(Oryctes rhinoceros, Linnaeus)。这种真菌是一种潜在的环境友好型生物制剂。本研究旨在从不同油棕种植土壤样品中分离出金银桃真菌,并利用RAPD标记分析其遗传多样性。本研究采用的方法是对绿僵菌(M. anisopliae, ITS)进行形态和分子鉴定,并利用RAPD标记进行遗传多样性检测。对北苏门答腊岛15个油棕种植地分离的15株绿僵菌进行了分子多样性分析。根据形态特征,所有分离株均鉴定为金龟子分枝杆菌。ITS引物测序结果表明,从土壤中分离得到的金龟子分枝杆菌与美国国家生物技术信息中心(NCBI)数据库中的金龟子分枝杆菌和布鲁氏分枝杆菌参考菌株同源性为85 ~ 99%。8条RAPD引物共生成39条可分带,其中38条(98.44%)为多态性条带。采用neighbor - joining Tree方法对RAPD图谱进行聚类分析,将RAPD图谱分成3组。关键词:昆虫病原真菌,遗传多样性,ITS测序,RAPD
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Keragaman genetik cendawan entomopatogen Metarhizium anisopliae (Metsch.) berasal dari tanah pertanaman kelapa sawit berdasarkan penanda RAPD
Metarizhium anisopliae (Metsch.) is an entomopathogenic fungus that attacks Oryctes rhinoceros (Linnaeus). This fungus is one of the potential bioagents that is environmentally friendly. The purpose of this study was to isolate the fungus M. anisopliae from various oil palm plantation soil samples and to see its genetic diversity based on RAPD markers. The method used in this research is the morphological and the molecular identification of M. anisopliae (ITS), and genetic diversity testing using RAPD markers. Fifteen isolates of M. anisopliae isolated from 15 oil palm plantation sites in North Sumatra were examined for their molecular diversity. On the basis of morphological characteristics, all isolates were identified as M. anisopliae. The result of sequencing with the ITS primer showed that all isolates of M. anisopliae isolated from soil had a 85–99% homology of with M. anisopliae and M. brunneum reference strains from gen bank database National Center Biotechnology Information (NCBI). Eight RAPD primers generated 39 scorable bands which 38 (98,44%) of them were polymorphic. Clustering analysis was performed based on RAPD profiles using Neighbour-Joining Tree method that formed 3 groups. Key words: entomopathogenic fungi, genetic diversity, ITS sequencing, RAPD
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