Yan Miaozhang, Li Lina, Wang Baoli, Qu Dong
College of Life Sciences, Northwest A&F University, Yangling 712100, China.
Wei Sheng Wu Xue Bao. 2013 Jun 4;53(6):577-85.
The diversity of Fe-hydrogenase based on Fe-hydrogenase gene of Clostridium was studied for exploring the biochemical mechanism of soil microbial hydrogen production and revealing the Fe-hydrogenase microbial community structure changes during the paddy soil flooding incubation.
We used denatured gradient gel electrophoresis and real-time quantitative PCR to achieve the goal.
The band number of Fe-hydrogenase denatured gradient gel electrophoresis fingerprints indicated Fe-hydrogenase microbes structure varied significantly during the completely flooding incubation. The Principal Component Analysis (PCA) showed the highly similar communities of Fe-hydrogenase microbial was divided into three groups: 1 d and 20 d, the 5 d, 30 d and 40 d. With the growth of the flooding incubation time Fe-hydrogenase microbial community structure became relatively stable and convergence. Alpha diversity index analysis found that the richness index (R), Shannon-Weaver index (H'), Simpson index (D(S)) numerical values of 1 d and 10 d were lower compared with other points, indicated the two time points of low Fe-hydrogenase diversity, simple Fe-hydrogenase microbial community structure and successive variation of community structure during the whole flooding incubation. After sequencing 15 Fe-hydrogenase preponderant bands ( labeled by G1 - G15), the phylogenetic tree of Fe-hydrogenase showed that the preponderant bands all had high similarity with the Clostridium Fe-hydrogenase in earlier flooding incubation stage and non-Clostridium Fe-hydrogenase in the later stage. Real-time quantitative PCR results demonstrated that the Fe-hydrogenase gene copy number was 10(6) level.
In the research we found 4 kinds of Clostridium Fe-hydrogenases and three kinds of non-Clostridium Fe-hydrogenases, the corresponding Fe-hydrogenase microbial community structure had successively significant variation in the early incubation stage and tend to be relatively stable and convergence in the late stage.
基于梭菌属铁氢化酶基因研究铁氢化酶的多样性，以探索土壤微生物产氢的生化机制，并揭示淹水培养过程中稻田土壤铁氢化酶微生物群落结构的变化。
我们采用变性梯度凝胶电泳和实时定量PCR来实现这一目标。
铁氢化酶变性梯度凝胶电泳指纹图谱的条带数量表明，在完全淹水培养期间，铁氢化酶微生物结构发生了显著变化。主成分分析（PCA）显示，铁氢化酶微生物群落高度相似，分为三组：1天和20天，5天、30天和40天。随着淹水培养时间的延长，铁氢化酶微生物群落结构变得相对稳定且趋同。α多样性指数分析发现，1天和10天的丰富度指数（R）、香农-威纳指数（H'）、辛普森指数（D(S)）数值低于其他时间点，表明这两个时间点铁氢化酶多样性较低，铁氢化酶微生物群落结构简单，且在整个淹水培养过程中群落结构连续变化。对15条铁氢化酶优势条带（标记为G1 - G15）进行测序后，铁氢化酶系统发育树表明，优势条带在淹水培养早期与梭菌属铁氢化酶高度相似，后期与非梭菌属铁氢化酶高度相似。实时定量PCR结果表明，铁氢化酶基因拷贝数为10(6)水平。
本研究发现4种梭菌属铁氢化酶和3种非梭菌属铁氢化酶，相应的铁氢化酶微生物群落结构在培养前期有显著的连续变化，后期趋于相对稳定且趋同。
