| 模拟干湿交替对水稻土古菌群落结构的影响 |
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| 引用本文： | 包丽君,贾仲君.模拟干湿交替对水稻土古菌群落结构的影响[J].土壤学报,2017,54(1):191-203. |
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| 作者姓名： | 包丽君 贾仲君 |
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| 作者单位： | 1. 中国科学院南京土壤研究所，南京 210008; 中国科学院大学，北京 100049;2. 中国科学院南京土壤研究所,南京,210008 |
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| 基金项目： | 国家重点基础研究发展计划（973计划）项目（2015CB150501） |
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| 摘 要： | 干湿交替是自然界普遍存在的现象,但长期以来由于技术的限制,复杂土壤中微生物对水分变化的响应规律仍不清楚。针对我国江苏常熟湖泊底泥发育的典型水稻土,在室内开展湿润-风干以及风干-湿润各三次循环,每次循环中湿润、风干状态各维持7d,利用微生物核糖体rRNA的通用引物进行PCR扩增,通过高通量测序分析土壤古菌多样性变化,同时结合实时荧光定量PCR技术,在DNA和RNA水平研究古菌数量对干湿交替过程的响应规律。结果表明:水稻土湿润-风干过程中,在DNA水平土壤古菌数量降幅约为149倍~468倍,而在RNA水平降幅最高仅为2.06倍;水稻土风干-湿润过程中,在DNA水平古菌数量增幅在147倍~360倍之间,而在RNA水平增幅最高仅为2.95倍。表明在干湿交替过程中,DNA水平的古菌16S rRNA基因数量变化远高于RNA水平。基于高通量测序多样性的结果表明,在DNA和RNA水平,湿润土壤3次风干、以及风干土壤3次加水湿润7d恢复后,土壤古菌群落结构均发生统计显著性改变。在微生物门、纲、目、科和属的不同分类水平下,水稻土古菌主要包括3、10、13、14、10种不同的类群,在RNA和DNA水平的结果基本一致。干湿交替导致部分古菌类群发生显著变化,其中在微生物分类学目水平发生显著变化的古菌最高达到6种,主要包括产甲烷古菌和氨氧化古菌,如Methanobacteriales、Methanosarcinales、Methanomicrobiales和Nitrososphaerales等。这些研究结果表明,反复的干湿交替并未显著改变水稻土中古菌的主要类群组成,古菌类群的绝对数量和相对丰度发生了一定程度的变化,但这些变化与微生物生理作用的联系仍需进一步研究;风干土壤中古菌RNA序列极可能来自于完整的古菌细胞,暗示了这些古菌细胞能够较好地适应水稻土中水分的剧烈变化,风干状态的土壤在一定程度也可用于土壤古菌群落组成研究。
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| 关 键 词： | 高通量测序 水稻土 16S rRNA 16S rRNA基因 干湿交替 古菌 |
| 收稿时间： | 2016/1/27 0:00:00 |
| 修稿时间： | 7/4/2016 12:00:00 AM |
Response of soil archaea communities to drying and rewetting cycles in paddy soil |
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| BAO Lijun and Jia Zhongjun.Response of soil archaea communities to drying and rewetting cycles in paddy soil[J].Acta Pedologica Sinica,2017,54(1):191-203. |
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| Authors: | BAO Lijun and Jia Zhongjun |
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| Institution: | Institute of Soil Science, Chinese Academy of Sciences and Institute of Soil Science, Chinese Academy of Sciences |
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| Abstract: | Objective]Dry-wet alternation is a common phenomenon existing in nature,especially in paddy soil. But it is still unclear about rule of the response of soil microorganisms to such variation of soil moisture in complex soil,because of limitations in technology. This study was aimed to investigate rules of changes inabundance and community of archaea in a typical paddy soil derived frnom lacustrine sediment, in Changshu of Jiangsu,experiencing three cycles of drying and wetting in lab. Method]Soil samples of the typical paddy soil from Changshu were subjected to three consective cycles of drying and wetting in lab. Each of the drying or wetting period lasted 7 days. Real-time quantitative polymerase chain reaction(Real-time PCR)and high-throughput sequencing of 16S rRNA genes were performed to analyze how the archaea in the soil samples changed in biodiversity,abundance and community in response to the dry-wet alternations at DNA and RNA levels. Result]The Real-time PCR analysis suggests that the abundance of archaeal 16S rRNA genes copy number changed in response to dry-wet alternation,dropping at DNA level by 149~468 times,but only by 2.06 times on RNA level during the process from wet to dry,and rising by 147~360 times at DNA level,but only by 2.95 times at RNA level during the process from dry to wet. These findings indicate that the change of archaea in 16S rRNA genes copy number was far greater at DNA level than at RNA level. Based on the high-throughput sequencing of 16S rRNA genes,it was found that the archaea community changed significantly in structure at both DNA level and RNA level after three consecutive dry-wet cycles and the changes may be described by non-metric multidimensional scaling(NMDS)(p<0.05). The archaea in the paddy soil could be sorted into 3,10,13,14 and 10 groups at either DNA or RNA level,when classified at phylum,class,order,family and genus level,respectively. The alternation caused significant changes in the archaeal community,especially the six groups at order level,including mainly Methanogenicarchaea and ammonia-oxidizing archaea,like Methanobacteriales,Methanosarcinales,Methanomicrobiales and Nitrososphaerales. The change varied by 2.1%,from 2.82% to 0.69% in total abundance and by 3.79 times, from 0.54% to 2.60%,in maximum fold. Conclusion]The Real-time PCR analysis demonstrates that the abundance of archaeal 16S rRNA genes copy number changed significantly in response to the alternation. The high-throughput sequencing of 16S rRNA genes indicates that the archaeal community in the soil changed significantly in structure with the soil alternating in soil moisture condition from dry to wet and from wet to dry. As in environment free extracellular RNAs decompose rapidly,while free extracellular DNAs may remain intact for quite a long time,it is quite probable that archaeal RNA sequence may come from intact microbial cells,and these archaeal cells are able to adapt to severe moisture changes in paddy soil. As the process of drying or wetting does have some impacts on abundance and composition of archaea communities,it is more advisable to unfold studies on changes in soil archaea at DNA and RNA levels simulataneously so as to expose rules of the soil archaea responding to dry-wet alternation of the soil. |
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| Keywords: | High-throughput sequencing Paddy soil 16S rRNA 16S rRNA gene Dry-wet alternation Archaea |
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