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广西山口红树林内生放线菌的分离、筛选及初步鉴定
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科技基础条件平台项目(No. 2005DKA21203); 国家自然科学基金项目(No. 30570040); “重大新药创制”科技重大专项(No. 2009ZX09301-003, 2009ZX09302-004)


Isolation, Screening and Preliminary Identification of Endophytic Actinobacteria from Mangroves at Shankou of Guangxi Province
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    摘要:

    从广西山口国家红树林生态自然保护区采集8种真红树植物和5种半红树植物的根、茎、叶及胚轴等组织样品共计41份。经过表面消毒后粉碎, 分别以几丁质、腐殖酸、木聚糖等作为主要营养及能量来源设计了8种分离培养基, 从植物组织中分离得到118株放线菌。使用6种新药筛选模型对分离菌株进行了生理活性筛选, 77株放线菌的发酵液样品在一个或多个药物筛选模型中显示为阳性, 初筛阳性率为65.3%。对77株筛选阳性菌株进行的初步分类研究结果显示, 其中44株属于链霉菌属, 25株为小单孢属的菌株, 3株属于糖丝菌属, 3株为诺卡氏属菌株, 1株是拟诺卡氏属菌株, 1株是伦茨氏菌属的菌株。对一株在抗多重耐药检定菌HH22 和降血脂模型上都显示出良好活性的菌株I07A-01824 进行表型和基因型分析, 确定I07A-01824为黄白链霉菌(Streptomyces albidoflavus)菌株。该研究显示, 红树植物内生环境不仅仅孕育了丰富多样的内生放线菌, 而且红树植物内生放线菌也是新型天然生理活性次级代谢产物的有效来源。

    Abstract:

    Fourty one plant tissue samples, including root, stem, leaf and hypocotyl were collected from 8 kinds of eumangrove and 5 kinds of semi-mangrove at Shankou Mangrove Nature Reserve in Guangxi Province. These samples were grinded into little pieces after surface sterilization, and then seeded into the isolation media containing chitin, humic acid or xylan as the main nutrition and energy source. As a result, 118 actinobacterial strains were purified from these plant tissue samples. These isolates were screened using 6 models for screening new antibiotics, and the fermentation broths from 77 strains exhibited activity on at least one screening model, the positive rate among the isolates is 65.3%. Based on the morphological characteristics, chemotaxonomic and 16S rRNA gene analysis, 77 isolates were identified as Streptomyces spp. (44), Micromonospora spp. (25), Saccharothrix spp. (3), Nocardia spp. (3), Nocardiopsis sp. (1) and Lentzea sp. (1). Strain I07A-01824, which showed strong antibacterial activity against the test strain Enterococcus faecalis HH22 (a multi-resistant clinical isolate) and positive activity on the screening model of statins-like antihyperlipidemics, was identified as a member of a known species Streptomyces albidoflavus. This study demonstrates that an unexpected diversity of actinobacteria colonizes inside the mangrove tissues, and these endophytic actinobacteria are promising sources of new natural secondary metabolites with physiological activities.

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魏玉珍,张玉琴,赵莉莉,李秋萍,苏静,刘红宇,孙承航,余利岩. 广西山口红树林内生放线菌的分离、筛选及初步鉴定[J]. 微生物学通报, 2010, 37(6): 0823-0828

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