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利用聚合酶链反应-变性梯度凝胶电泳技术鉴定巴里恩托斯岛南极土壤中的放线菌群落。

Identification of actinomycete communities in Antarctic soil from Barrientos Island using PCR-denaturing gradient gel electrophoresis.

作者信息

Learn-Han L, Yoke-Kqueen C, Shiran M S, Vui-Ling C M W, Nurul-Syakima A M, Son R, Andrade H M

机构信息

Department of Biomedical Science, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang, Selangor Darul Ehsan, Malaysia.

出版信息

Genet Mol Res. 2012 Feb 8;11(1):277-91. doi: 10.4238/2012.February.8.3.

DOI:10.4238/2012.February.8.3
PMID:22370930
Abstract

The diversity of specific bacteria taxa, such as the actinomycetes, has not been reported from the Antarctic island of Barrientos. The diversity of actinomycetes was estimated with two different strategies that use PCR-denaturing gradient gel electrophoresis. First, a PCR was applied, using a group-specific primer that allows selective amplification of actinomycete sequences. Second, a nested-PCR approach was used that allows the estimation of the relative abundance of actinomycetes within the bacterial community. Molecular identification, which was based on 16S rDNA sequence analysis, revealed eight genera of actinomycetes, Actinobacterium, Actinomyces, an uncultured Actinomycete, Streptomyces, Leifsonia, Frankineae, Rhodococcus, and Mycobacterium. The uncultured Actinomyces sp and Rhodococcus sp appear to be the prominent genera of actinomycetes in Barrientos Island soil. PCR-denaturing gradient gel electrophoresis patterns were used to look for correlations between actinomycete abundance and environmental characteristics, such as type of rookery and vegetation. There was a significant positive correlation between type of rookery and abundance of actinomycetes; soil samples collected from active chinstrap penguin rookeries had the highest actinomycete abundance. Vegetation type, such as moss, which could provide a microhabitat for bacteria, did not correlate significantly with actinomycete abundance.

摘要

在巴连托斯南极岛屿,尚未有关于特定细菌类群(如放线菌)多样性的报道。采用两种不同的利用聚合酶链反应-变性梯度凝胶电泳的策略对放线菌的多样性进行了评估。首先,应用聚合酶链反应,使用一组特异性引物,实现对放线菌序列的选择性扩增。其次,采用巢式聚合酶链反应方法,以估计细菌群落中放线菌的相对丰度。基于16S核糖体DNA序列分析的分子鉴定揭示了8个放线菌属,即放线杆菌属、放线菌属、一种未培养的放线菌、链霉菌属、利夫森菌属、弗兰克氏菌科、红球菌属和分枝杆菌属。未培养的放线菌属和红球菌属似乎是巴连托斯岛土壤中放线菌的主要属。利用聚合酶链反应-变性梯度凝胶电泳图谱来寻找放线菌丰度与环境特征(如繁殖地类型和植被)之间的相关性。繁殖地类型与放线菌丰度之间存在显著正相关;从活跃的南极帽带企鹅繁殖地采集的土壤样本中放线菌丰度最高。植被类型(如苔藓)虽可为细菌提供微生境,但与放线菌丰度并无显著相关性。

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