新疆土壤VBNC状态菌的复苏可培养化及其系统发育关系研究
发布时间：2018-11-21 15:45
【摘要】：新疆土壤由于环境特殊,人为干扰较少,拥有巨大的微生物资源宝库,但是目前通过传统培养方法能分离到的生境中的菌种不到总量的0.01%-10%,大部分微生物资源因为处在活的不可培养(viable but non-culturable,VBNC)状态而未被人们认识,更无法被我们所用。VBNC状态菌作为自然界中未能开发利用的巨大微生物资源,新的微生物资源的鉴定及微生物环境功能的研究值得引起关注。本课题旨在基于复苏促进因子(1resuscitation promoting factor, Rpf),采用MPN(most probable number)培养体系,对新疆土壤样品进行VBNC资源菌的复苏可培养化研究。共获得并分离新疆土壤菌株60株,其中VBNC状态菌48株,构建了一个小型的VBNC资源菌库,丰富了实验室的菌种资源。所分离VBNC菌株分属于16个属：Bacillus, Brevibacillus, Staphylococcus, Brevibacterium, Arthrobacter, Kocuria, Microbacterium, Cellulosimicrobium, Rhodococcus, Streptomyces, Enterobacter, Escherichia, Brevundimonas, Pseudomonas, Ensifer 和 Herbaspirillum。体现了新疆土壤Rpf复苏可培养化的VBNC菌具有分布广,种类多的特性。其中以Bacillus属菌数最多,占分离VBNC菌株总数的40%；放线菌属种类最多,包括Streptomyces, Microbacterium等7个菌属。经过对分离菌株的初步分类学鉴定,共获得14株潜在的新种。通过形态学、细胞化学和分子等多相分类学手段对其中一株Arthrobacter属的菌株DSXY973完成了菌株系统发育定位和描述工作。本研究表明Rpf不仅对高GC革兰氏阳性菌有良好的复苏促进作用,对低GC革兰氏阳性菌也具有很好的复苏促进效果,其中Bacillus属对Rpf最为敏感,另外,Rpf对一部分革兰氏阴性菌也有一定的复苏促进效果。本研究为Rpf对环境中VBNC状态资源菌的复苏分离提供了科学依据,为新菌种资源的开发和后续应用研究提供了新的思路和途径。
[Abstract]:Because of the special environment and less human disturbance, Xinjiang soil has a huge treasure trove of microbial resources, but the bacteria species in the habitats that can be separated by traditional culture methods are less than 0.01-10% of the total amount. The majority of microbial resources are not known because they are in a living, uncultivated (viable but non-culturable,VBNC state, and they are not used by us as a huge microbial resource that cannot be exploited and utilized in nature. The identification of new microbial resources and the study of microbial environmental function deserve attention. The purpose of this study was to study the resuscitation and culturability of VBNC resource bacteria in Xinjiang soil samples based on 1resuscitation promoting factor, Rpf), (MPN (most probable number) culture system). A total of 60 strains of soil strains were obtained and isolated from Xinjiang, of which 48 strains were VBNC status bacteria. A small library of VBNC resources was constructed, which enriched the resources of bacteria in the laboratory. The isolated VBNC strains belong to 16 genera: Bacillus, Brevibacillus, Staphylococcus, Brevibacterium, Arthrobacter, Kocuria, Microbacterium, Cellulosimicrobium, Rhodococcus, Streptomyces, Enterobacter, Escherichia, Brevundimonas, Pseudomonas, Ensifer and Herbaspirillum.. The results showed that the VBNC bacteria which could be cultivated by soil Rpf resuscitation in Xinjiang had the characteristics of wide distribution and variety. Among them, the Bacillus genus was the largest, accounting for 40% of the total isolated VBNC strains, and the actinomycetes genus was the most, including 7 genera such as Streptomyces, Microbacterium. Through the preliminary taxonomic identification of the isolated strains, a total of 14 potential new species were obtained. The phylogenetic localization and description of one of the strains of Arthrobacter genus DSXY973 were carried out by means of morphological, cytochemistry and molecular multiphasic taxonomy. The results showed that Rpf not only promoted the recovery of Gram-positive bacteria with high GC, but also promoted the resuscitation of Gram-positive bacteria with low GC. Bacillus genus was the most sensitive to Rpf. Rpf can also promote the recovery of some Gram-negative bacteria. This study provides a scientific basis for the recovery and isolation of VBNC resource bacteria in the environment by Rpf, and provides new ideas and approaches for the development and application of new strain resources.
【学位授予单位】：浙江师范大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S154.3
本文编号：2347411
[Abstract]:Because of the special environment and less human disturbance, Xinjiang soil has a huge treasure trove of microbial resources, but the bacteria species in the habitats that can be separated by traditional culture methods are less than 0.01-10% of the total amount. The majority of microbial resources are not known because they are in a living, uncultivated (viable but non-culturable,VBNC state, and they are not used by us as a huge microbial resource that cannot be exploited and utilized in nature. The identification of new microbial resources and the study of microbial environmental function deserve attention. The purpose of this study was to study the resuscitation and culturability of VBNC resource bacteria in Xinjiang soil samples based on 1resuscitation promoting factor, Rpf), (MPN (most probable number) culture system). A total of 60 strains of soil strains were obtained and isolated from Xinjiang, of which 48 strains were VBNC status bacteria. A small library of VBNC resources was constructed, which enriched the resources of bacteria in the laboratory. The isolated VBNC strains belong to 16 genera: Bacillus, Brevibacillus, Staphylococcus, Brevibacterium, Arthrobacter, Kocuria, Microbacterium, Cellulosimicrobium, Rhodococcus, Streptomyces, Enterobacter, Escherichia, Brevundimonas, Pseudomonas, Ensifer and Herbaspirillum.. The results showed that the VBNC bacteria which could be cultivated by soil Rpf resuscitation in Xinjiang had the characteristics of wide distribution and variety. Among them, the Bacillus genus was the largest, accounting for 40% of the total isolated VBNC strains, and the actinomycetes genus was the most, including 7 genera such as Streptomyces, Microbacterium. Through the preliminary taxonomic identification of the isolated strains, a total of 14 potential new species were obtained. The phylogenetic localization and description of one of the strains of Arthrobacter genus DSXY973 were carried out by means of morphological, cytochemistry and molecular multiphasic taxonomy. The results showed that Rpf not only promoted the recovery of Gram-positive bacteria with high GC, but also promoted the resuscitation of Gram-positive bacteria with low GC. Bacillus genus was the most sensitive to Rpf. Rpf can also promote the recovery of some Gram-negative bacteria. This study provides a scientific basis for the recovery and isolation of VBNC resource bacteria in the environment by Rpf, and provides new ideas and approaches for the development and application of new strain resources.
【学位授予单位】：浙江师范大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S154.3
【参考文献】
相关期刊论文 前1条
1 张英,李伟,王妙冬,施耀;微杆菌ZD-M2降解二苯并噻吩的特性及其生长条件优化[J];化工学报;2005年07期
，本文编号：2347411
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