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作物杂志,2017, 第1期: 127–134 doi: 10.16035/j.issn.1001-7283.2017.01.023

• 生理生化·植物营养·栽培耕作 • 上一篇    下一篇

不同水分条件对玉米根际微生物群落的影响

冯帅1,2,刘小利1,2,吴小丽1,2,江世杰2,周正富2,张维2,陈明2,王劲1,2,柯秀彬2   

  1. 1西南科技大学生命科学与工程学院,621010,四川绵阳
    2中国农业科学院生物技术研究所,100081,北京
  • 收稿日期:2016-11-10 修回日期:2016-12-18 出版日期:2017-02-15 发布日期:2018-08-26
  • 作者简介:冯帅,硕士研究生,研究方向为农业微生物基因组学
  • 基金资助:
    国家973计划(2015CB755701);国家自然科学基金(31400439);转基因重大专项(2014ZX0800301B)

Effects of Water Conditions on Microbial Community in Maize Rhizosphere

Feng Shuai1,2,Liu Xiaoli1,2,Wu Xiaoli1,2,Jiang Shijie2,Zhou Zhengfu2,Zhang Wei2,Chen Ming2,Wang Jin1,2,Ke Xiubin2   

  1. 1Life Science and Engineering College,Southwest University of Science and Technology,Mianyang 621010,Sichuan
    2Biotechnology Research Institute,Chinese Academy of Agriculture Science,Beijing 100081,China
  • Received:2016-11-10 Revised:2016-12-18 Online:2017-02-15 Published:2018-08-26

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摘要：

通过玉米盆栽试验研究不同水分条件下（土壤孔隙含水量20%和80%）土壤微生物群落结构及种群数量的变化。采集玉米生长第60天时的根际土和表层土,通过MiSeq Illumina高通量测序技术扩增细菌和古菌16S rRNA基因分析群落多样性,同时通过绝对定量PCR扩增细菌和古菌16S rRNA基因分析其种群数量。结果表明：高通量测序共得到41 546个细菌OTUs和6 921个古菌OTUs。土壤中细菌的多样性丰富,优势菌群为变形菌（Proteobacteria）和酸杆菌（Acidobacteria）,而古菌的群落组成单一,优势菌群为奇古菌（Thaumarchaeota）。非度量多维尺度（NMDS）分析表明不同土壤区域中细菌群落结构具有明显差异,而古菌群落结构受水分和土壤区域的影响不明显。定量分析结果显示细菌和古菌数量受土壤区域和水分条件的影响显著。表层土中细菌和古菌数量均显著高于根际土。根际土中细菌和古菌数量在水分充足条件下均显著高于干旱条件,表层土中细菌和古菌数量也表现出相同趋势。本研究结果表明土壤不同区域（生态位）可能是影响土壤中微生物群落最重要的因素。

关键词: 玉米根际, 微生物群落, 细菌, 古菌, 16S rRNA多样性测序, qPCR

Abstract:

Maize microcosms were constructed to investigate the dynamics of soil microbial community structure and population at different water treatments (20% and 80% WFPS) under greenhouse condition. We quantified the abundance and characterized the community structures based on the 16S rRNA gene of bacteria and archaea, using qPCR and barcoded pyrosequencing, by collecting the rhizosphere soil and surface soil at the 60th day of maize growth. Results showed that 41 546 OTUs of bacteria and 6 921 OTUs of archaea were obtained by high throughput sequencing. Higher diversity of bacteria was observed, and it was dominant by Proteobacteria and Acidobacteria. While the community structure of archaea was low, and it was predominant by Thaumarchaeota. Nonmetric Multidimensional Scaling (NMDS) analysis showed a significant difference in bacterium community pattern between rhizosphere soil and surface soil, but archaea community structure was not significantly affected. The abundances of bacterium and archaea were also significantly affected by “soil compartments”. Higher abundance of bacterium and archaea were found in surface soil than rhizosphere soil. In the rhizosphere soil, the abundance of bacterium and archaea under well-watered condition was significantly higher than those under drought condition. Besides, bacterium and archaea showed the similar trend in the surface soil. Our study indicates that soil compartments (niche differentiation) may be the most important factor affecting microbial community in agricultural soil.

Key words: Maize rhizosphere, Microbial community, Bacterium, Archaea, 16S rRNA pyrosequencing, Quantitative PCR

图1

第60天时玉米生长量 数值为平均值±标准误（n=3）,不同字母表示差异显著（P<0.05）,下同"

图2

第60天玉米根际土和表层土中细菌与古菌16S rRNA基因拷贝数"

表1

方差分析环境因子（土壤区域、水分条件）对细菌和古菌16S rRNA基因拷贝数的影响"

变异来源Source of variation 自由度
df		 细菌Bacterium 古菌Archaea
F值F-value P(>F) F值F-value P(>F)
土壤区域Soil compartment 1 87.39 <0.01 127.90 <0.01
水分条件Water treatment 1 54.21 <0.01 79.67 <0.01
土壤区域×水分条件Soil compartment×Water treatment 1 28.89 <0.01 50.79 <0.01

表2

细菌16S rRNA基因多样性指数"

水分条件Water treatment 土壤区域Soil Compartment 操作分类单元OTUs 丰富度指数Chao1 多样性指数Shannon index 测序深度指数Coverage
干旱Water stress 根际土Rhizosphere soil 10 337 19 030.3±172.1 10.8±0.03 0.966
水充足Well-watered 根际土Rhizosphere soil 9 074 20 662.5±97.7 10.9±0.02 0.986
干旱Water stress 表层土Surface soil 9 839 20 438.0±546.3 10.7±0.06 0.994
水充足Well-watered 表层土Surface soil 12 296 16 311.5±815.2 10.1±0.14 0.995

表3

古菌16s rRNA基因多样性指数"

水分条件Water treatment 土壤区域Soil Compartment 操作分类单元OTUs 丰富度指数Chao1 多样性指数Shannon index 测序深度指数Coverage
干旱Water stress 根际土Rhizosphere soil 1 252 229.2±33.7 4.3±0.10 0.996
水充足Well-watered 根际土Rhizosphere soil 1 710 272.3±26.8 4.5±0.05 0.959
干旱Water stress 表层土Surface soil 2 320 300.7±29.3 4.8±0.03 0.979
水充足Well-watered 表层土Surface soil 1 639 186.2±16.2 4.5±0.05 0.979

表4

玉米所有根际土和表层土中细菌16S rRNA基因文库的前10个OTUs及注释"

OTU号码
OTU number		 分类Taxon 相对丰度 (%)
Relative abundance
OTU_18078 f_ Anaerolineaceae 3.85
OTU_24643 g_ Salinimicrobium 3.28
OTU_5453 f_ Cytophagaceae 1.86
OTU_26825 g_ Planktothrix 1.65
OTU_1926 f_ Anaerolineaceae 1.53
OTU_10853 s_ Arthrobacter globiformis 0.81
OTU_47774 g_ Flavisolibacter 0.65
OTU_35806 g_ Ramlibacter 0.63
OTU_43589 g_ Marmoricola 0.53
OTU_56073 c_ Acidobacteria 0.53

表5

玉米所有根际土和表层土中古菌16S rRNA基因文库的前10位OTUs及注释"

OTU号码
OTU number		 分类Taxon 相对丰度(%)
Relative abundance
OTU_41732 c_ Soil Crenarchaeotic Group 31.89
OTU_18108 c_ Soil Crenarchaeotic Group 6.64
OTU_44564 c_ Soil Crenarchaeotic Group 5.79
OTU_48728 c_ Soil Crenarchaeotic Group 5.14
OTU_47697 c_ Soil Crenarchaeotic Group 4.31
OTU_53860 c_ Soil Crenarchaeotic Group 2.77
OTU_3112 c_ Soil Crenarchaeotic Group 1.54
OTU_3179 c_ Soil Crenarchaeotic Group 1.32
OTU_53911 c_ Soil Crenarchaeotic Group 1.25
OTU_28469 c_ Soil Crenarchaeotic Group 1.09

图3

在纲分类水平上根际土和表层土中细菌群落结构与古菌群落结构"

图4

非度量多维尺度分析土壤区域（生态位）、水分条件因素对细菌和古菌群落结构的影响"

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