Discipline of Marine Biotechnology and Ecology, CSIR-Central Salt and Marine Chemicals Research Institute (CSMCRI), G. B. Marg, Bhavnagar 364002, Gujarat, India.
Microbiol Res. 2013 Mar 30;168(3):165-73. doi: 10.1016/j.micres.2012.09.005. Epub 2012 Oct 16.
Population indices of bacteria and archaea were investigated from saline-alkaline soil and a possible microbe-environment pattern was established using gene targeted metagenomics. Clone libraries were constructed using 16S rRNA and functional gene(s) involved in carbon fixation (cbbL), nitrogen fixation (nifH), ammonia oxidation (amoA) and sulfur metabolism (apsA). Molecular phylogeny revealed the dominance of Actinobacteria, Firmicutes and Proteobacteria along with archaeal members of Halobacteraceae. The library consisted of novel bacterial (20%) and archaeal (38%) genera showing ≤95% similarity to previously retrieved sequences. Phylogenetic analysis indicated ability of inhabitant to survive in stress condition. The 16S rRNA gene libraries contained novel gene sequences and were distantly homologous with cultured bacteria. Functional gene libraries were found unique and most of the clones were distantly related to Proteobacteria, while clones of nifH gene library also showed homology with Cyanobacteria and Firmicutes. Quantitative real-time PCR exhibited that bacterial abundance was two orders of magnitude higher than archaeal. The gene(s) quantification indicated the size of the functional guilds harboring relevant key genes. The study provides insights on microbial ecology and different metabolic interactions occurring in saline-alkaline soil, possessing phylogenetically diverse groups of bacteria and archaea, which may be explored further for gene cataloging and metabolic profiling.
采用基于基因的宏基因组学技术研究了盐碱土中的细菌和古菌的种群指数,并建立了可能的微生物环境模式。使用 16S rRNA 和参与碳固定(cbbL)、氮固定(nifH)、氨氧化(amoA)和硫代谢(apsA)的功能基因构建了克隆文库。分子系统发育分析表明,放线菌、厚壁菌门和变形菌门以及盐杆菌科的古菌成员占优势。该文库由新的细菌(20%)和古菌(38%)组成,与之前检索到的序列相似度≤95%。系统发育分析表明,这些微生物具有在应激条件下生存的能力。16S rRNA 基因文库包含新的基因序列,与培养的细菌具有远缘同源性。功能基因文库是独特的,大多数克隆与变形菌门关系较远,而 nifH 基因文库的克隆也与蓝细菌和厚壁菌门有同源性。定量实时 PCR 显示,细菌丰度比古菌高两个数量级。基因定量表明了携带相关关键基因的功能群的大小。该研究提供了关于微生物生态学和盐碱土中发生的不同代谢相互作用的见解,这些微生物具有系统发育上多样化的细菌和古菌群体,可以进一步探索其基因分类和代谢分析。