“使它隆”对玉米根部不同微生环境细菌群落多样性的影响
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摘要
【目的】研究除草剂"使它隆"施用后对玉米根部不同微生环境细菌群落结构和多样性的影响。【方法】利用Illumina Miseq高通量测序技术,对玉米根系内生菌、根际和非根际土壤细菌16S rRNA的V4–V5可变区序列进行测定,分析不同生长期喷施除草剂使它隆对玉米土壤细菌及根系内生菌群落结构和多样性的影响。【结果】本研究15个样品共得到544393条有效序列,333565条优质序列。多样本共有OTU分析表明,非根际和根际土壤的群落结构更为相似,在一定程度上说明玉米根部相关细菌的定殖具有选择性并且是从根际到根系逐步专一化。丰度等级曲线和Alpha多样性结果显示非根际和根际土壤群落的丰富度和均匀度较高,而玉米根系内生菌群落的丰富度和均匀度都比较低,且成熟期玉米根系内生菌群落的丰富度在施用除草剂使它隆后下降比较剧烈。群落组成分析发现,使它隆除草后,玉米根部相关细菌各时期在门及属水平上的分布都发生了较大变化。菌群代谢功能预测结果表明玉米生长从苗期到成熟期,微生物的生长压力逐渐加大,需要消耗更多的能量用于新陈代谢和环境适应。【结论】施用除草剂使它隆后会降低玉米根部土壤细菌群落的多样性,使它隆对成熟期玉米根系内生菌群落影响最为显著。
[Objective] The purpose of this research is to study the effects of herbicide "starane" on the structure and diversity of bacterial communities in different micro-environments. [Methods] We used the high-throughput sequencing technique Illumina Miseq to determine the V4–V5 variable region sequence of 16 S rRNA in endophytic, rhizosphere and non-rhizosphere soils of maize roots. The herbicides were sprayed in different growth stages, and we researched the herbicide effect on the structure and diversity of maize root endophytes as well as soil bacteria. [Results] We obtained a total of 544393 effective sequences and 333565 high-quality sequences from 15 samples. The multi-sample OTU analysis showed that the community structure of non-rhizosphere soil and rhizosphere soil was similar, which indicated that the colonization of maize-related bacteria was selective and the gradual specialization from rhizosphere to root. The results of rank abundance curve and Alpha diversity showed that the richness and uniformity of non-rhizosphere and rhizosphere soil were higher, while the richness and uniformity of endophytic community in maize roots of mature stage were lower. The abundance of endophytic community in maize roots was significantly reduced by the application of herbicides. The composition analysis of the community showed that the distribution of bacteria in the maize roots was with a great change at both the Phylum and Genus level. The prediction of metabolic function of the flora indicated that the growth pressure of the microorganisms was gradually increased from the seeding stage to the mature stage of the corn, which required more energy for bacterial metabolism and environmental adaptation. [Conclusion] The application of herbicide "starane" can reduce the diversity of bacterial community in the roots of maize, and the effect on the endophytic community of maize roots is more obvious.
[Objective] The purpose of this research is to study the effects of herbicide "starane" on the structure and diversity of bacterial communities in different micro-environments. [Methods] We used the high-throughput sequencing technique Illumina Miseq to determine the V4–V5 variable region sequence of 16 S rRNA in endophytic, rhizosphere and non-rhizosphere soils of maize roots. The herbicides were sprayed in different growth stages, and we researched the herbicide effect on the structure and diversity of maize root endophytes as well as soil bacteria. [Results] We obtained a total of 544393 effective sequences and 333565 high-quality sequences from 15 samples. The multi-sample OTU analysis showed that the community structure of non-rhizosphere soil and rhizosphere soil was similar, which indicated that the colonization of maize-related bacteria was selective and the gradual specialization from rhizosphere to root. The results of rank abundance curve and Alpha diversity showed that the richness and uniformity of non-rhizosphere and rhizosphere soil were higher, while the richness and uniformity of endophytic community in maize roots of mature stage were lower. The abundance of endophytic community in maize roots was significantly reduced by the application of herbicides. The composition analysis of the community showed that the distribution of bacteria in the maize roots was with a great change at both the Phylum and Genus level. The prediction of metabolic function of the flora indicated that the growth pressure of the microorganisms was gradually increased from the seeding stage to the mature stage of the corn, which required more energy for bacterial metabolism and environmental adaptation. [Conclusion] The application of herbicide "starane" can reduce the diversity of bacterial community in the roots of maize, and the effect on the endophytic community of maize roots is more obvious.
引文
[1]Bu B,Yang JZ,Gong GS,Zhang M,Liu W,Song XY.Effect of plastic film mulch and herbicides on weeds and maize growth in a summer field.Weed Science,2013,31(3):40-43.(in Chinese)补彬,杨继芝,龚国淑,张敏,刘武,宋小宇.地膜覆盖和除草剂对夏玉米田杂草及玉米生长发育的影响.杂草学报,2013,31(3):40-43.
[2]Wang NN.The study of the community structures of endophytes within common plant species and soil microorganisms in Inner Mongolia grassland.Master Dissertation of Jiangxi Agricultural University,2013.(in Chinese)王纳纳.内蒙古草原典型植物内生菌和土壤微生物群落结构的研究.江西农业大学硕士学位论文,2013.
[3]Sheng Y.Effects of herbicide chlorimuron-ethyl on soil microbial ecology.Master Dissertation of Northeast Agricultural University,2011.(in Chinese)盛宇.长残留除草剂氯嘧磺隆对土壤微生物生态的影响.东北农业大学硕士学位论文,2011.
[4]Song XD.The study of safety and weed control effect of hormone herbicide on the naked oats.Master Dissertation of Gansu Agricultural University,2015.(in Chinese)宋旭东.激素型除草剂对裸燕麦的安全性及其除草效果研究.甘肃农业大学硕士学位论文,2015.
[5]Xu YR,Fang ZJ,Lu XP,Hao LJ.Effects of starane on maize soil bacterial diversity analyzed by high-throughput sequencing technology.Acta Microbiologica Sinica,2017,57(7):985-993.(in Chinese)许艳蕊,方志军,卢晓平,郝鲁江.基于高通量测序技术分析使它隆对玉米土壤细菌多样性的影响.微生物学报,2017,57(7):985-993.
[6]Wang CL,Fang ZJ,Xu YR,Lu XP,Mu CH,Shan K,Hao LJ.Effects of starane on the community diversity of maize root endophytes analyzed using high-throughput sequencing technology.Crops,2018,(1):160-165.(in Chinese)王春蕾,方志军,许艳蕊,卢晓平,穆春华,单凯,郝鲁江.基于高通量测序技术分析使它隆对玉米根系内生菌多样性的影响.作物杂志,2018,(1):160-165.
[7]Zhao F,Zhao MZ,Wang Y,Pang FH.Biodiversity of bacteria and fungi in rhizosphere of strawberry with different continuous cropping years.Microbiology China,2017,44(6):1377-1386.(in Chinese)赵帆,赵密珍,王钰,庞夫花.不同连作年限草莓根际细菌和真菌多样性变化.微生物学通报,2017,44(6):1377-1386.
[8]Mago?T,Salzberg SL.FLASH:fast length adjustment of short reads to improve genome assemblies.Bioinformatics,2011,27(21):2957-2963.
[9]Caporaso JG,Kuczynski J,Stombaugh J,Bittinger K,Bushman FD,Costello EK,Fierer N,Pe?a AG,Goodrich JK,Gordon JI,Huttley GA,Kelley ST,Knights D,Koenig JE,Ley RE,Lozupone CA,McDonald D,Muegge BD,Pirrung M,Reeder J,Sevinsky JR,Turnbaugh PJ,Walters WA,Widmann J,Yatsunenko T,Zaneveld J,Knight R.QIIME allows analysis of high-throughput community sequencing data.Nature Methods,2010,7(5):335-336.
[10]Edgar RC.Search and clustering orders of magnitude faster than BLAST.Bioinformatics,2010,26(19):2460-2461.
[11]Langille MGI,Zaneveld J,Caporaso JG,McDonald D,Knights D,Reyes JA,Clemente JC,Burkepile DE,Thurber RLV,Knight R,Beiko RG,Huttenhower C.Predictive functional profiling of microbial communities using 16SrRNA marker gene sequences.Nature Biotechnology,2013,31(9):814-821.
[12]Ramette A.Multivariate analyses in microbial ecology.FEMSMicrobiology Ecology,2007,62(2):142-160.
[13]Fang Y,Wang W,Yao XD,Peng XQ.Soil microbial community composition and environmental controls in northern temperate steppe of China.Acta Scientiarum Naturalium Universitatis Pekinensis,2017,53(1):142-150.(in Chinese)方圆,王娓,姚晓东,彭晓茜.我国北方温带草地土壤微生物群落组成及其环境影响因素.北京大学学报(自然科学版),2017,53(1):142-150.
[14]Yu J.Diversity of root endophytic bacteria and identification of promoting growth bacteria in corn and soybean.Doctor Dissertation of Northeast Agricultural University,2016.(in Chinese)喻江.玉米和大豆根内生细菌多样性及促生细菌鉴定评价.东北农业大学博士学位论文,2016.
[15]Wang XC.The influence of long term rice-rice-green manure rotation on rice bulk soil and root-associated bacteria.Master Dissertation of Chinese Academy of Agricultural Sciences,2015.(in Chinese)王秀呈.稻-稻-绿肥长期轮作对水稻土壤及根系细菌群落的影响.中国农业科学院硕士学位论文,2015.
[16]Cui YS.Bacterial community structure and function in rhizospheric soil of Panax notoginseng.Master Dissertation of Kunming University of Science and Technology,2017.(in Chinese)崔尹赡.三七根际土壤细菌的群落结构与功能研究.昆明理工大学硕士学位论文,2017.
[17]Yao B,Zhang CL.Effect of three herbicides on microbial biomass C,N and respiration in paddy soil.Ecology and Environment,2008,17(2):580-583.(in Chinese)姚斌,张超兰.除草剂对土壤微生物生物量碳、氮及呼吸的影响.生态环境,2008,17(2):580-583.
[18]Sun SQ,Liu X,Yao Y,Gao ZG,Sun YQ,Li A.Effects of atrazine and nicosulfuron on soil microorganism.Agrochemicals,2014,53(4):276-279.(in Chinese)孙淑清,刘限,姚远,高增贵,孙艳秋,李安.莠去津和烟嘧磺隆对玉米田土壤微生物的影响.农药,2014,53(4):276-279.
[19]Li X,Zhang JY,Zhang MM,Ju CM,Zheng GY,Zhang HH,Cai DJ,Hu JW,Sun GY.Effects of chemical herbicide at different spraying ways on the carbon source metabolism of soil microbial community in alfalfa rhizosphere.Acta Agrestia Sinica,2014,22(1):57-64.(in Chinese)李鑫,张景云,张萌萌,鞠成梅,郑桂英,张会慧,蔡敦江,胡举伟,孙广玉.化学除草剂不同施用方法对紫花苜蓿根际土壤微生物群落碳源利用的影响.草地学报,2014,22(1):57-64.
[2]Wang NN.The study of the community structures of endophytes within common plant species and soil microorganisms in Inner Mongolia grassland.Master Dissertation of Jiangxi Agricultural University,2013.(in Chinese)王纳纳.内蒙古草原典型植物内生菌和土壤微生物群落结构的研究.江西农业大学硕士学位论文,2013.
[3]Sheng Y.Effects of herbicide chlorimuron-ethyl on soil microbial ecology.Master Dissertation of Northeast Agricultural University,2011.(in Chinese)盛宇.长残留除草剂氯嘧磺隆对土壤微生物生态的影响.东北农业大学硕士学位论文,2011.
[4]Song XD.The study of safety and weed control effect of hormone herbicide on the naked oats.Master Dissertation of Gansu Agricultural University,2015.(in Chinese)宋旭东.激素型除草剂对裸燕麦的安全性及其除草效果研究.甘肃农业大学硕士学位论文,2015.
[5]Xu YR,Fang ZJ,Lu XP,Hao LJ.Effects of starane on maize soil bacterial diversity analyzed by high-throughput sequencing technology.Acta Microbiologica Sinica,2017,57(7):985-993.(in Chinese)许艳蕊,方志军,卢晓平,郝鲁江.基于高通量测序技术分析使它隆对玉米土壤细菌多样性的影响.微生物学报,2017,57(7):985-993.
[6]Wang CL,Fang ZJ,Xu YR,Lu XP,Mu CH,Shan K,Hao LJ.Effects of starane on the community diversity of maize root endophytes analyzed using high-throughput sequencing technology.Crops,2018,(1):160-165.(in Chinese)王春蕾,方志军,许艳蕊,卢晓平,穆春华,单凯,郝鲁江.基于高通量测序技术分析使它隆对玉米根系内生菌多样性的影响.作物杂志,2018,(1):160-165.
[7]Zhao F,Zhao MZ,Wang Y,Pang FH.Biodiversity of bacteria and fungi in rhizosphere of strawberry with different continuous cropping years.Microbiology China,2017,44(6):1377-1386.(in Chinese)赵帆,赵密珍,王钰,庞夫花.不同连作年限草莓根际细菌和真菌多样性变化.微生物学通报,2017,44(6):1377-1386.
[8]Mago?T,Salzberg SL.FLASH:fast length adjustment of short reads to improve genome assemblies.Bioinformatics,2011,27(21):2957-2963.
[9]Caporaso JG,Kuczynski J,Stombaugh J,Bittinger K,Bushman FD,Costello EK,Fierer N,Pe?a AG,Goodrich JK,Gordon JI,Huttley GA,Kelley ST,Knights D,Koenig JE,Ley RE,Lozupone CA,McDonald D,Muegge BD,Pirrung M,Reeder J,Sevinsky JR,Turnbaugh PJ,Walters WA,Widmann J,Yatsunenko T,Zaneveld J,Knight R.QIIME allows analysis of high-throughput community sequencing data.Nature Methods,2010,7(5):335-336.
[10]Edgar RC.Search and clustering orders of magnitude faster than BLAST.Bioinformatics,2010,26(19):2460-2461.
[11]Langille MGI,Zaneveld J,Caporaso JG,McDonald D,Knights D,Reyes JA,Clemente JC,Burkepile DE,Thurber RLV,Knight R,Beiko RG,Huttenhower C.Predictive functional profiling of microbial communities using 16SrRNA marker gene sequences.Nature Biotechnology,2013,31(9):814-821.
[12]Ramette A.Multivariate analyses in microbial ecology.FEMSMicrobiology Ecology,2007,62(2):142-160.
[13]Fang Y,Wang W,Yao XD,Peng XQ.Soil microbial community composition and environmental controls in northern temperate steppe of China.Acta Scientiarum Naturalium Universitatis Pekinensis,2017,53(1):142-150.(in Chinese)方圆,王娓,姚晓东,彭晓茜.我国北方温带草地土壤微生物群落组成及其环境影响因素.北京大学学报(自然科学版),2017,53(1):142-150.
[14]Yu J.Diversity of root endophytic bacteria and identification of promoting growth bacteria in corn and soybean.Doctor Dissertation of Northeast Agricultural University,2016.(in Chinese)喻江.玉米和大豆根内生细菌多样性及促生细菌鉴定评价.东北农业大学博士学位论文,2016.
[15]Wang XC.The influence of long term rice-rice-green manure rotation on rice bulk soil and root-associated bacteria.Master Dissertation of Chinese Academy of Agricultural Sciences,2015.(in Chinese)王秀呈.稻-稻-绿肥长期轮作对水稻土壤及根系细菌群落的影响.中国农业科学院硕士学位论文,2015.
[16]Cui YS.Bacterial community structure and function in rhizospheric soil of Panax notoginseng.Master Dissertation of Kunming University of Science and Technology,2017.(in Chinese)崔尹赡.三七根际土壤细菌的群落结构与功能研究.昆明理工大学硕士学位论文,2017.
[17]Yao B,Zhang CL.Effect of three herbicides on microbial biomass C,N and respiration in paddy soil.Ecology and Environment,2008,17(2):580-583.(in Chinese)姚斌,张超兰.除草剂对土壤微生物生物量碳、氮及呼吸的影响.生态环境,2008,17(2):580-583.
[18]Sun SQ,Liu X,Yao Y,Gao ZG,Sun YQ,Li A.Effects of atrazine and nicosulfuron on soil microorganism.Agrochemicals,2014,53(4):276-279.(in Chinese)孙淑清,刘限,姚远,高增贵,孙艳秋,李安.莠去津和烟嘧磺隆对玉米田土壤微生物的影响.农药,2014,53(4):276-279.
[19]Li X,Zhang JY,Zhang MM,Ju CM,Zheng GY,Zhang HH,Cai DJ,Hu JW,Sun GY.Effects of chemical herbicide at different spraying ways on the carbon source metabolism of soil microbial community in alfalfa rhizosphere.Acta Agrestia Sinica,2014,22(1):57-64.(in Chinese)李鑫,张景云,张萌萌,鞠成梅,郑桂英,张会慧,蔡敦江,胡举伟,孙广玉.化学除草剂不同施用方法对紫花苜蓿根际土壤微生物群落碳源利用的影响.草地学报,2014,22(1):57-64.