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| 施氮水平对稻-稻-紫云英稻田土壤细菌数量及群落结构的影响 |
| Effect of nitrogen application level on abundance and community structure of paddy soil bacteria under ricerice-Chinese milk vetch(Astragalus sinicus L.)cropping system |
| 投稿时间：2018-01-17 |
| DOI：10.13254/j.jare.2018.0021 |
| 中文关键词: 紫云英，氮肥，双季稻，细菌群落结构，高通量测序，荧光定量 |
| 英文关键词: vetch, nitrogen fertilizer, double cropping rice, soil bacterial community structure, high-throughput sequencing, real-time PCR |
| 基金项目:国家自然科学基金项目（31501274，31870424）；中国科协青年人才托举工程项目（YESS20160040）；湿地生态与农业利用教育部工程研究中心2015年开放基金项目（KF201504） |
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| 中文摘要: |
| 通过对稻-稻-紫云英稻田土壤细菌数量和群落结构多样性进行研究，揭示不同施氮水平对稻田土壤微生物的影响，以期为促进稻田土壤养分管理提供依据。以湖南省8年定位田间试验稻田土壤为对象，试验处理以冬闲水稻田不施氮肥为对照（CK），设置冬种紫云英还田条件下水稻季三个施氮肥水平分别为不施氮肥（0 kg N·hm-2，CN0）、施中水平氮肥（100 kg N·hm-2，CN100）、施高水平氮肥（200 kg N·hm-2，CN200）。采用荧光定量PCR和Illumina MiSeq高通量测序平台分别研究了不同处理下稻田土壤的总细菌数量和群落结构。结果表明：稻田土壤总细菌数量为1.25×108~8.47×109拷贝数·g-1干土；处理间物种多样性指数（Shannon和Simpson）和物种丰富度指数（Ace和Chao）均存在显著性差异；16S rRNA基因在门水平分类下3个主要类群是变形菌门（Proteobacteria）、绿弯菌门（Chloroflexi）和硝化螺旋菌门（Nitrospirae），分别占总OTU比例的42.22%~54.54%、8.56%~20.73%、10.98%~15.08%；CN100土壤样品中变形菌门的相对丰度低于CN200、CN0和CK，绿弯菌门的相对丰度分别为CN200、CN0和CK的2.26、1.58倍和1.17倍。相关性分析表明，土壤细菌16S rRNA基因拷贝数与铵态氮含量呈显著正相关。影响土壤细菌菌群结构的因子分析结果表明，土壤细菌与土壤pH值、铵态氮、硝态氮存在密切相关性。研究表明，在冬种紫云英还田条件下施加氮肥可以显著改变湖南省双季稻区土壤微生物数量与结构。 |
| 英文摘要: |
| In order to study the effect of nitrogen rate on soil microbial ecology and provide a basis for the promotion of soil nutrient management in paddy fields, we investigated the abundance and community structure of paddy soil bacteria in a rice-rice-vetch cropping system. Soil samples were taken from an 8-year continuous field trial, with three treatments(no nitrogen application, 0 kg N·hm-2, CN0; middle nitrogen application level, 100 kg N·hm-2, CN100; high nitrogen application level, 200 kg N·hm-2, CN200) under milk vetch returning condition, and fallow was set as control(CK). Soil bacterial abundance and community structure were estimated by using fluorescence quantitative PCR and Illumina MiSeq high-throughput sequencing platform, respectively. The results showed that the abundance of soil bacteria ranged from 1.25×108 to 8.47×109 copies per gram dry soil in all soil samples. Sequencing results indicated that community diversity indexes(Shannon and Simpson) and community richness indexes(Ace and Chao) had significant variation among different treatments at 0.05 probability level. Proteobacteria, Chloroflexi and Nitrospirae were three mainly soil bacterial groups under Phylum classification level, accounting for total proportion of OTU 42.22%~54.54%, 8.56%~20.73% and 10.98%~15.08%, respectively. The relative abundance of Proteobacteria of soils from CN100 was lower than those from the other three treatments, while Chloroflexi relative abundance was 2.26, 1.58 and 1.17 times higher than CN200, CN0 and CK, respectively. Correlation analysis showed that bacterial 16S rRNA gene copy numbers was positively correlated with soil ammonium nitrogen. Factors analysis of soil bacterial community structure indicated that there was a close correlation between soil bacteria and soil chemical indicators. A forementioned results demonstrated that soil microbial quantity and community structure could be significantly altered by nitrogen application combined with vetch incorporation in double cropping soils. |
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