淹水时间对水稻土中地杆菌科群落结构及丰度的影响

【目的】通过模拟水稻土淹水过程,探讨地杆菌科(Geobacteraceae)群落结构和相对丰度随淹水时间的动态变化特征,揭示其群落结构和相对丰度变化与微生物Fe(Ⅲ)还原的内在联系。【方法】提取水稻土淹水培养1 h、1 d、5 d、10 d、20 d和30 d后的微生物总DNA,构建地杆菌科16S rDNA克隆文库,采用PCR-RFLP方法分析地杆菌科的群落结构和多样性变化特征,通过Real-time PCR技术测定地杆菌科相对丰度的动态变化。采用厌氧泥浆培养方法,测定水稻土中Fe(Ⅱ)产生量变化。【结果】供试水稻土中,微生物Fe(Ⅲ)还原过程在淹水培养初期变化明显,培养20 d后达到稳定期,最大铁还原潜势为10.16 mg/g,最大反应速率为1.064 mg/(g.d),最大反应速率对应的时间为4.84 d。α多样性指数显示,水稻土中地杆菌科的多样性随淹水时间延长呈现波动性变化,淹水5 d和20 d处理出现2个峰值,而淹水10 d和30 d处理的多样性明显减小。β多样性指数表明淹水过程中群落结构存在明显差异。不同淹水时间共产生了10种地杆菌科优势类型,分别属于Clade 1和Clade 2。Real-time PCR结果表明,地杆菌科与总细菌16S rDNA丰度的比值在淹水培养1 d时最小(1.20%),而20 d时达到最大值(4.54%)。【结论】淹水培养的水稻土中,地杆菌科微生物的多样性和相对丰度的动态变化与微生物Fe(Ⅲ)还原过程密切相关。

Effect of flooding time on community structure and abundance of Geobacteraceae in paddy soil

Abstract: Objective] The dynamic characteristics of community structure and relative abundance of Geobacteraceae were investigated to understand their response to microbial iron(III) reducing in flooded paddy soil. Methods] The paddy soil was incubated anaerobically and the amount of Fe(II) was determined during the flooding incubation. We retrieved Geobacteraceae sequences from clone libraries constructed for different time points (1 h and day 1, 5, 10, 20 and 30) after flooding of the paddy soil. The diversity and community structure were analyzed by using RFLP method, and the relative abundance of Geobacteraceae was detected by real-time PCR. Results] Microbial reduction of iron(III) changed greatly in early time and was stable after incubated for 20 d in paddy soil. The largest iron reduction potential was 10.16 mg/g with a Vmax of 1.064 mg/(g.d) at the time of 4.84 d whereas this process achieved plateau after 20 days flooding. Diversity of Geobacteraceae, given by alpha indices, fluctuated during the flooding incubation. Two peaks of diversity were observed in treatments of 5 d and 20 d respectively, while significant low diversity appeared in samples of 10 d and 30 d. Beta indices described the differences between community structures of Geobacteraceae and hence reflected the variation of the flooding situation over time. In all samples, 10 RFLP-based preponderant types were found, which fell into clade 1 and clade 2 of Geobacteraceae. The relative abundance of Geobacteraceae was the lowest in 1 d (1.20%) and the highest in 20 d (4.54%). Conclusion] The dynamic variation of Geobacteraceae diversity, community structure and abundance are closely related to microbial iron(III) reducing in flooding paddy soil.