sRNA在鞘氨醇单胞菌TY降解尼古丁中的调控功能研究

微生物降解是去除环境中尼古丁的主要途径。sRNA作为广泛存在于细菌内且对基因表达起重要调控作用的调控元件获得了越来越多的关注。目前sRNA研究主要在致病菌致病机制和模式菌株压力应答方面，sRNA调控污染物降解及胁迫条件下污染物降解的研究还十分缺乏。鞘氨醇单胞菌TY可高效降解尼古丁，课题组前期对其降解分子机制进行了深入研究。本研究拟以菌株TY为研究对象，在前期基础上，利用RNA-seq和生物信息分析等技术预测与尼古丁降解及低温胁迫下尼古丁降解相关的sRNA，通过RT-qPCR、Northern blot、基因敲除、基因回补与过表达等技术鉴定其功能，进一步利用EMSA、融合子活性检测、点突变及RNA二级结构鉴定等手段阐明sRNA调控尼古丁降解的机制。本研究有助于深入了解在污染物降解过程中以及低温情况下sRNA的调控情况，对于微生物在生物反应器、土壤强化修复及污水处理中的低温瓶颈有重要意义。

Microbial bioremediation is the main way to relieve the nicotine residue in environment. As the most important regulatory elements found in many bacteria, sRNAs paly important role in physiology activities and gained growing attention in recent years. Many researches about sRNA have been focused on the pathogenesis of pathogenic bacteria or stress response of several type strains, little attention has been paid to pollutant degradation and regulation of pollutant degradation under adversity. Sphingomonas melonis TY was an effective nicotine-degrading strain. Besides, the pathway and the molecular mechanism of nicotine degradation in strain TY have been fully studied by our research group. Based on the previous study, we will use the data analysis of sRNA RNA-seq to predict putative nicotine degradation- and nicotine degradation under cold stress- related sRNA first, then by using RT-qPCR, Northern blot, gene knockout and gene complementation to identity the role of these predicted sRNA, and using the EMSA, activity of infusions, point mutation and secondary structure of RNA to clarify the regulatory function of sRNA in nicotine degradation. Our research will help to understand the function of sRNA in pollutant degradation and their regulatory role in pollutant degradation to resist stress. In addition, this study will provide important guidance to the bioreactor, soil bioremediation, and waste water treatment, especially to the low activity under cold environment.