干旱区生态环境知识资源中心(Arid): 醉马草免培养内生细菌的多样性

Arid

	醉马草免培养内生细菌的多样性
其他题名	Endophytic bacterial diversity in Achnatherum inebrians by culture-independent approach
	张雪兵; 史应武; 曾军; 娄恺
来源期刊	生态学报
ISSN	1000-0933
出版年	2011
卷号	31 期号:8 页码:2178-2187
中文摘要	为了解醉马草内生细菌的组成和多样性,采用液氮研磨法提取醉马草总DNA,利用内生细菌16S rRNA基因特异性引物对醉马草总DNA进行16S rRNA基因扩增,构建醉马草内生细菌16S rRNA基因文库.对筛选到249个克隆进行HaeШ酶切分析,得到57个操作分类单元(OTUs).系统发育分析将其归为4个门:变形杆菌门(Proteobacteria),厚壁菌门(Firmicutes),放线菌门(Actinobacteria),拟杆菌门(Bacteroidetes)以及1个未知类群.其中74%的克隆与可培养细菌中的37个属具有高的16SrRNA基因序列相似性(97%--100%),其中芽孢杆菌属(Bacillus spp.),红球菌属(Rhodococcus spp.),黄杆菌属(Flavobacterium spp.),黄单胞杆菌属(Xanthomonas spp.)最为丰富.另外26%的克隆序列与GenBank中已存细菌16S rRNA基因序列相似性小于96%.研究结果表明,醉马草中可培养内生细菌丰富,多样并且可能存在一些潜在新物种或新类群
英文摘要	Achnatherum inebrians(Hance) Keng, which is belonged to genus Achnatherum Beauv, Gramineae family, also known as Oxytropis glabra,is one of poisonous perennial herbs on grassland and is wildly distributed in Inner Mongolia, Ningxia, Gansu, Qinghai and Xinjiang province of northwest China. It is toxic to hoofed mammals of the genus Equus, however, they are hardy and easy to grow in arid and semi-arid areas and played active role in improving ecological environment. The toxic ingredients extracted from the plant are usually used as anti-cancer drugs, immune modulators;such as Swainsonine, one of well studied anti-cancer drugs that often used to stimulate bone marrow cell proliferates. Therefore, the plant is worth of fullly usage and utilizing it rationally. In this study,a culture-independent of analysis were chosen to investigate the endophytic bacterial composition and diversity associated in Achnatherum inebrians. Total community DNA were extracted by modified CTAB-SDS DNA extraction protocol from plant stem, leaf and root which were grinded up using liquid nitrogen grinding method after strictly surface-sterilization. To construct 16S rDNA clone libraries, endophytic bacterial specific primer pair 799F and 1492R were selected to amplify bacterial 16S rDNA partial gene, and then PCR products were ligated into a PMD18-T Vector and transformed into Escherichia coli DH 5alpha. Transformants were selected by blue and white selection. Positive clones were analyzed by Restriction Fragment length polymorphism(RFLP), and clones with unique RFLP patterns were selected for sequencing, alignment and constructing 16S rRNA gene phylogenetic tree. In total 249 clones were screened, which could be grouped into 57 operational taxonomic units(OTUs), based on Hae III digestion. These were divided into four phyla(Proteobacteria, Firmicutes, Actinobacteria, Bacteroides) and one unknown group by phylogenetic analysis. Of them,74% of clones classified into 37 genera of bacteria phylum were highly related to cultivable species with high 16S rRNA gene similarity(>96% sequence similarity);and among them genera Bacillus spp., Rhodococcus spp., Flavobacterium spp., Xanthomonas spp., were most abundant;In addition,26% of clones showed less than 96% of 16S rRNA gene identity with bacterial sequences deposited in GenBank database. Besides, rarefaction curve reaches clear saturation and its Coverage C value is 93%, indicating that most of the estimated endophytic bacterial diversity was sampled and 249 positive clones for enzyme digestion were enough good to the community investigation. These results suggest that Achnatherum inebrians harbors a diversity of cultivable endophytic bacteria and maybe existence of some potential novel species or lineages
中文关键词	醉马草 ; 内生细菌 ; 免培养法 ; 生物多样性
英文关键词	Achnatherum inebrians endophytic bacterial culture-independent biodiversity
语种	中文
国家	中国
收录类别	CSCD
WOS类目	ECOLOGY
WOS研究方向	Environmental Sciences & Ecology
CSCD记录号	CSCD:4182715
资源类型	期刊论文
条目标识符	http://119.78.100.177/qdio/handle/2XILL650/226908
作者单位	新疆农业科学院微生物应用研究所, 乌鲁木齐, 新疆 830091, 中国
推荐引用方式 GB/T 7714	张雪兵,史应武,曾军,等. 醉马草免培养内生细菌的多样性[J],2011,31(8):2178-2187.
APA	张雪兵,史应武,曾军,&娄恺.(2011).醉马草免培养内生细菌的多样性.生态学报,31(8),2178-2187.
MLA	张雪兵,et al."醉马草免培养内生细菌的多样性".生态学报 31.8(2011):2178-2187.