云南三种金属尾矿中的原核微生物群落特征及重金属耐受菌株和基因的挖掘

Ore resource of Yunnan province is abundant. With minerals processing, masses of low grade tailings were discarded. Heavy metals in tailings destroyed environment greatly, which threatens human health. Bio-leaching is an important tool for tailings treatment. However, the screening of bioleaching microorganism is important because of low leaching rate and affected easily by external factors. The shortcut is obtaining heavy-metal tolerant microorganism from tailings. In our previous study, we isolated some heavy-metal tolerant bacteria from tailings in Yunnan. In this project, we aim to analyze relationship between Prokaryote communities and heavy-metals by comparing Prokaryote communities in Gejiu tin tailing, Dongchuan copper-titanium tailing and Lanping Jinding Lead-Zinc tailing using 16S rRNA gene (V3-V4 region for bacteria and V4 region for archaea) high-throughput sequencing technology. Prokaryote community differences between inside and outside the tailings can illuminate the change law of Prokaryote community. Through isolating the Prokaryotes, detecting their metal-resistance genes (Zhf, PvSR2, merA and nixA) and tolerance against temperature, pH, heavy metals, and metal-resistance Prokaryotes will be obtained. The performance of this project can provide new Prokaryote resources and reference data for understanding the Prokaryote community structure and bioleaching systems.

云南有着丰富的金属矿产资源。矿山开采形成的尾矿严重破坏了生态环境，给人类健康带来隐患。用微生物对低品位尾矿浸出是消除重金属污染的重要渠道之一。但浸出率低、易受环境条件影响等特点使得浸矿菌种的选育尤为重要，研究表明从尾矿中可筛选获得高效浸矿菌种。我们前期已从尾矿中分离了一些重金属抗性菌株。基于此，本课题拟通过16S rRNA基因（细菌V3-V4区，古菌V4区）高通量测序分析云南个旧锡矿、东川铜钛矿、兰坪金顶铅锌矿三种尾矿中的原核微生物群落结构，揭示不同金属尾矿的微生物群落特点，比较尾矿内外微生物群落组成差异，分析重金属对微生物群落的影响；分离尾矿原核微生物，检测其重金属抗性、抗性基因（Zhf, PvSR2, merA和nixA）、温度和低pH耐受等特征，为深入理解不同金属尾矿中的微生物群落特征及生理特性提供数据，同时为处理尾矿、治理重金属污染提供参考和可筛选的菌种/基因资源库。

云南有着丰富的金属矿产资源。矿山开采形成的尾矿严重破坏了生态环境，给人类健康带来隐患。用微生物对低品位尾矿浸出是消除重金属污染的重要渠道之一。但浸出率低、易受环境条件影响等特点使得浸矿菌种的选育尤为重要，研究表明从尾矿中可筛选获得高效浸矿菌种。我们前期已从尾矿中分离了一些重金属抗性菌株。本项目根据研究路线，通过分析16S rRNA基因高通量测序分析云南个旧锡矿、东川铜钛矿、兰坪金顶铅锌矿三种尾矿中的原核微生物群落结构发现，Acitinobacter、Proteobacteria、Acidobacteriota、Chloroflexi、Gemmatimonadota、Bacteroidota、Firmicutes、Myxococcota、Methylomirabilota、Patescibacteria是尾矿区的主要原核微生物类群。其中，个旧尾矿的Acitinobacter类群数量是兰坪和东川的近两倍，而Firmicutes却是兰坪和东川的不到20%；尾矿内的重金属含量远远高于尾矿外，比较尾矿内外微生物群落组成差异发现，尾矿内 微生物类群丰度远远低于矿外。通过尾矿原核微生物分离，共获得826株菌（去重复后），其中筛选到了9株高抗重金属同时耐低pH的放线菌菌株；获得了Zhf, PvSR2, merA和nixA基因呈阳性的菌株163株。本研究结果为深入理解不同金属尾矿中的微生物群落特征及生理特性提供了数据，同时为处理尾矿、治理重金属污染提供了参考和可筛选的菌种/基因资源库。