水稻条斑病抗感品种根际微生物群落结构和功能分析
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摘要
通过抗感水稻细菌性条斑病(简称细条病)的品种的水根际可培养细菌群落结构和代谢功能多样性分析,探讨其根际细菌多样性差异与水稻抗细条病的相关性。采集孕穗期水稻品种CG2和IR24的植株根际土壤,采用多种培养基进行可培养细菌的分离、鉴定,并通过16S rRNA序列构建系统进化树;利用Biolog技术分析不同品种根际微生物的代谢功能差异。结果显示:不同水稻品种的根际土壤可培养细菌分析发现抗病品种CG2以芽孢杆菌为优势菌群,而感病品种IR24以节杆菌为优势种群;对条斑病菌有抑制的菌株,筛选后以芽孢杆菌为主,且抗病品种的比率达47%以上,而感病材料中只有2.3%;抗病品种根际微生物群落的Shannon多样性指数(H′), Shannon均匀度指数(SE), Simpson指数(D), McIntos指数(U)和McIntosh均匀度指数(ME)分别增加了8.80%, 48.49%, 11.76%,41.88%和0.52%;主成份分析表明抗病品种CG2的根际土壤微生物与感病品种IR24相比,对碳水化合物,氨基酸类、酯类、醇类、胺类和酸类的利用率分别提高了12.52%、66.53%、3.78%、7.49%、27.98%和40.67%。研究表明,从根际土壤细菌数量和种类上看,抗病品种CG2的根际土壤细菌数量和种类与感病品种相比,细菌群落结构多样性更为丰富,发挥生防作用的菌群更多,为水稻条斑病的生物防治提供了思路和资源。
We analyzed the rice rhizosphere microbiome composition and metabolism of different resistance rice cultivars, to explore the relationship between the rhizosphere soil microbial diversity and resistance to rice bacterial leaf streak. We collected the rhizosphere soil of plants CG2 and IR24 rice varieties in booting stage, using a variety of culture medium to cultivate bacteria isolation, purification, counting, and through the 16 S rRNA homology comparison and phylogenetic tree analysis, and microbial metabolic function difference was studied by using Biolog. The results showed that 1057 single colony different varieties were isolated from the culture medium, and they were divided into 15 types, 6 genera: Bacillus,Arthrobacter, Chryseobacterium, Sphingobacterium, Microbacterium, Curtobacterium. Plate count in rhizosphere soil of different rice varieties of culturable bacteria showed bacillus was the dominant species in resistance cultivar CG2 and Arthrobacter was dominant species in the susceptible cultivar IR24. Shannon diversity index(H) and Shannon evenness index(SE), Simpson index(D), McIntos index(U) and McIntosh evenness index(ME) of the rhizosphere microbial community of resistance cultivar CG2 were increased by 8.80%, 48.49%, 11.76%, 41.88% and 0.52%, as compared to the susceptible cultivar IR24. The principal component analysis showed that the use of carbohydrates, amino acids, esters,alcohols, amine and acid were increased by 12.52%, 66.53%, 3.78%, 7.49%, 27.98%, and 40.67% for the resistance cultivar CG2 rhizosphere soil microorganisms as compared with susceptible variety IR24. The research shows that the colony number and kind of the resistant cultivars CG2 rhizosphere soil bacteria are more than the susceptible cultivars IR24. The bacterial diversity is more abundant; the community structure is more stable; niche competition is stronger than pathogenic bacteria.The results can provide ideas and resources for the biological control of rice bacterial leaf streak.
We analyzed the rice rhizosphere microbiome composition and metabolism of different resistance rice cultivars, to explore the relationship between the rhizosphere soil microbial diversity and resistance to rice bacterial leaf streak. We collected the rhizosphere soil of plants CG2 and IR24 rice varieties in booting stage, using a variety of culture medium to cultivate bacteria isolation, purification, counting, and through the 16 S rRNA homology comparison and phylogenetic tree analysis, and microbial metabolic function difference was studied by using Biolog. The results showed that 1057 single colony different varieties were isolated from the culture medium, and they were divided into 15 types, 6 genera: Bacillus,Arthrobacter, Chryseobacterium, Sphingobacterium, Microbacterium, Curtobacterium. Plate count in rhizosphere soil of different rice varieties of culturable bacteria showed bacillus was the dominant species in resistance cultivar CG2 and Arthrobacter was dominant species in the susceptible cultivar IR24. Shannon diversity index(H) and Shannon evenness index(SE), Simpson index(D), McIntos index(U) and McIntosh evenness index(ME) of the rhizosphere microbial community of resistance cultivar CG2 were increased by 8.80%, 48.49%, 11.76%, 41.88% and 0.52%, as compared to the susceptible cultivar IR24. The principal component analysis showed that the use of carbohydrates, amino acids, esters,alcohols, amine and acid were increased by 12.52%, 66.53%, 3.78%, 7.49%, 27.98%, and 40.67% for the resistance cultivar CG2 rhizosphere soil microorganisms as compared with susceptible variety IR24. The research shows that the colony number and kind of the resistant cultivars CG2 rhizosphere soil bacteria are more than the susceptible cultivars IR24. The bacterial diversity is more abundant; the community structure is more stable; niche competition is stronger than pathogenic bacteria.The results can provide ideas and resources for the biological control of rice bacterial leaf streak.
引文
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[13]李洪连,袁红霞,王烨,等.根际微生物多样性与棉花品种对黄萎病抗性关系研究--Ⅰ.根际微生物数量与棉花品种对黄萎病抗性的关系[J].植物病理学报,1998(04):341-345.
[14]蔡秋华,左进香,李忠环,等.抗性烤烟品种根际微生物数量及功能多样性差异[J].应用生态学报,2015,26(12):3766-3772.
[15]雷娟利,寿伟松,董文其,等.抗感枯萎病西瓜根际微生物比较研究[J].微生物学通报,2008,(07):1034-1038.
[16]陈宏宇,李晓鸣,王敬国.抗病性不同大豆品种根面及根际微生物区系的变化Ⅰ.非连作大豆(正茬)根面及根际微生物区系的变化[J].植物营养与肥料学报,2005,(06):98-103+109.
[17]ADAME-áLVAREZ R,MENDIOLA-SOTO J,HEIL M.Order of arrival shifts endophyte-pathogen interactions in bean from resistance induction to disease facilitation[J].FEMS microbiology letters,2014,355(2):100-107.
[18]D’ALESSANDRO M,ERB M,TON J,et al.Volatiles produced by soil-borne endophytic bacteria increase plant pathogen resistance and affect tritrophic interactions[J].Plant,cell&environment,2014,37(4):813-826.
[19]SPENCE C,ALFF E,JOHNSON C,et al.Natural rice rhizospheric microbes suppress rice blast infections[J].BMC plant biology,2014,14(1):130.
[20]陆晓菊,官会林,张正芸,等.三七连作根际土壤微生物区系的16S rRNA系统遗传多样性[J].微生物学报,2015,55(02):205-213.
[21]WEBER K,GROVE J,GEHDER M,et al.Data transformations in the analysis of community-level substrate utilization data from microplates[J].Journal of Microbiological Methods,2007,69(3):461-469.
[22]董艳,董坤,汤利,等.小麦蚕豆间作对蚕豆根际微生物群落功能多样性的影响及其与蚕豆枯萎病发生的关系[J].生态学报,2013,33(23):7445-7454.
[23]CHOI K,DOBBS F.Comparison of two kinds of Biolog microplates(GN and ECO)in their ability to distinguish among aquatic microbial communities[J].Journal of Microbiological Methods,1999,36(3):203-213.
[24]熊君,林辉锋,李振方,等.旱直播条件下强弱化感潜力水稻根际微生物的群落结构[J].生态学报,2012,32(19),6100-6109.
[25]潘丽媛,肖炜,董艳,等.超高产生态区水稻根际微生物物种及功能多样性研究[J].农业资源与环境学报,2016,33(6):583-590.
[26]YANTI Y.Peroxidase Enzyme Activity of RhizobacteriaIntroduced Shallots Bulbs to Induce Resistance of Shallot towards Bacterial Leaf Blight(Xanthomonas axonopodis pv allii)[J].Procedia Chemistry,2015,14(1):501-507.
[27]赵明富,李梦娇,张芬芬,等.变栖克雷伯菌在石斛体内的定殖动态及其对石斛黑斑病的防效试验[J].西南林业大学学报,2015,35(03):14-19.
[28]任慧爽,徐伟芳,王爱印,等.桑树内生细菌多样性及内生拮抗活性菌群的研究[J].西南大学学报(自然科学版),2017,39(01):36-45.
[29]田新莉,曹理想,杨国武,等.水稻内生放线菌类群及其对宿主病原菌的抗性研究[J].微生物学报,2004,(05):641-646.
[2]SWARTHOUT D,HARPER E,JUDD S,et al.Measures of leaf-level water-use efficiency in drought stressed endophyte infected and non-infected tall fescue grasses[J].Environmental and Experimental Botany,2009.66(1),88-93.
[3]DE L F,LEIA C,FERNANDA L,ARAúJO W.Epicoccum nigrum P16,a sugarcane endophyte,produces antifungal compounds and induces root growth[J].PLoS One,2012,7(6):e36826.
[4]SINGH M,SINGH D,MESAPOGU S,et al.Concomitant colonization of nifH positive endophytic Burkholderia sp.in rice(Oryza sativa L.)promotes plant growth[J].World Journal of Microbiology and Biotechnology,2011,27(9):2023-2031.
[5]EDWARDS J,JOHNSON C,SANTOS-MEDELLíN C,et al.Structure,variation,and assembly of the root-associated microbiomes of rice[J].Proceedings of the National Academy of Sciences,2015,112(8):E911-E920.
[6]THIJS S,VAN DILLEWIJN P,SILLEN W,et al.Exploring the rhizospheric and endophytic bacterial communities of Acer pseudoplatanus growing on a TNT-contaminated soil:towards the development of a rhizocompetent TNT-detoxifying plant growth promoting consortium[J].Plant and soil,2014,385(1-2):15-36.
[7]赵丹,赵娟,刘涛,等.元阳梯田水稻品种‘月亮谷’根系内生及根际细菌的群落结构[J].云南农业大学学报(自然科学),2017,32(01):17-26.
[8]MENDES L,KURAMAE E,NAVARRETE A,et al.Taxonomical and functional microbial community selection in soybean rhizosphere[J].The ISME journal,2014,8(8):1577-1587.
[9]YAN Y,KURAMAE E,DE HOLLANDER M,et al.Functional traits dominate the diversity-related selection of bacterial communities in the rhizosphere[J].The ISMEjournal,2017,11(1):56-66.
[10]PII Y,MIMMO T,TOMASI N,et al.Microbial interactions in the rhizosphere:beneficial influences of plant growthpromoting rhizobacteria on nutrient acquisition process.Areview[J].Biology and fertility of soils,2015,51(4):403-415.
[11]CHAPARRO J,BADRI D,VIVANCO J.Rhizosphere microbiome assemblage is affected by plant development[J].The ISME journal,2014,8(4):790-803.
[12]CARVALHAIS L C,DENNIS P G,BADRI D V,et al.Linking jasmonic acid signaling,root exudates,and rhizosphere microbiomes[J].Molecular Plant-Microbe Interactions,2015,28(9):1049-1058.
[13]李洪连,袁红霞,王烨,等.根际微生物多样性与棉花品种对黄萎病抗性关系研究--Ⅰ.根际微生物数量与棉花品种对黄萎病抗性的关系[J].植物病理学报,1998(04):341-345.
[14]蔡秋华,左进香,李忠环,等.抗性烤烟品种根际微生物数量及功能多样性差异[J].应用生态学报,2015,26(12):3766-3772.
[15]雷娟利,寿伟松,董文其,等.抗感枯萎病西瓜根际微生物比较研究[J].微生物学通报,2008,(07):1034-1038.
[16]陈宏宇,李晓鸣,王敬国.抗病性不同大豆品种根面及根际微生物区系的变化Ⅰ.非连作大豆(正茬)根面及根际微生物区系的变化[J].植物营养与肥料学报,2005,(06):98-103+109.
[17]ADAME-áLVAREZ R,MENDIOLA-SOTO J,HEIL M.Order of arrival shifts endophyte-pathogen interactions in bean from resistance induction to disease facilitation[J].FEMS microbiology letters,2014,355(2):100-107.
[18]D’ALESSANDRO M,ERB M,TON J,et al.Volatiles produced by soil-borne endophytic bacteria increase plant pathogen resistance and affect tritrophic interactions[J].Plant,cell&environment,2014,37(4):813-826.
[19]SPENCE C,ALFF E,JOHNSON C,et al.Natural rice rhizospheric microbes suppress rice blast infections[J].BMC plant biology,2014,14(1):130.
[20]陆晓菊,官会林,张正芸,等.三七连作根际土壤微生物区系的16S rRNA系统遗传多样性[J].微生物学报,2015,55(02):205-213.
[21]WEBER K,GROVE J,GEHDER M,et al.Data transformations in the analysis of community-level substrate utilization data from microplates[J].Journal of Microbiological Methods,2007,69(3):461-469.
[22]董艳,董坤,汤利,等.小麦蚕豆间作对蚕豆根际微生物群落功能多样性的影响及其与蚕豆枯萎病发生的关系[J].生态学报,2013,33(23):7445-7454.
[23]CHOI K,DOBBS F.Comparison of two kinds of Biolog microplates(GN and ECO)in their ability to distinguish among aquatic microbial communities[J].Journal of Microbiological Methods,1999,36(3):203-213.
[24]熊君,林辉锋,李振方,等.旱直播条件下强弱化感潜力水稻根际微生物的群落结构[J].生态学报,2012,32(19),6100-6109.
[25]潘丽媛,肖炜,董艳,等.超高产生态区水稻根际微生物物种及功能多样性研究[J].农业资源与环境学报,2016,33(6):583-590.
[26]YANTI Y.Peroxidase Enzyme Activity of RhizobacteriaIntroduced Shallots Bulbs to Induce Resistance of Shallot towards Bacterial Leaf Blight(Xanthomonas axonopodis pv allii)[J].Procedia Chemistry,2015,14(1):501-507.
[27]赵明富,李梦娇,张芬芬,等.变栖克雷伯菌在石斛体内的定殖动态及其对石斛黑斑病的防效试验[J].西南林业大学学报,2015,35(03):14-19.
[28]任慧爽,徐伟芳,王爱印,等.桑树内生细菌多样性及内生拮抗活性菌群的研究[J].西南大学学报(自然科学版),2017,39(01):36-45.
[29]田新莉,曹理想,杨国武,等.水稻内生放线菌类群及其对宿主病原菌的抗性研究[J].微生物学报,2004,(05):641-646.