健康与发病香蕉植株内生细菌菌群差异
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摘要
[目的]本研究通过探究健康和发病‘巴西香牙蕉’植株根、茎、叶内生细菌群落结构差异,为生防菌筛选和香蕉枯萎病的防控工作提供参考。[方法]通过MiSeq高通量测序细菌16S核糖体RNA分析香蕉枯萎病发病蕉园健康和发病‘巴西香牙蕉’(Musa AAA cavendish cv. Brazil)植株根、茎、叶内生细菌群落结构与多样性。[结果]健康‘巴西香牙蕉’植株根内生细菌菌群多样性和丰富度指数最高分别是6.11和671,其次为叶,分别是5.49和370,最后是茎,分别是2.50和237;发病‘巴西香牙蕉’植株的根内生细菌菌群多样性和丰富度指数分别为6.86和1 226,茎部内生细菌菌群多样性和丰富度指数分别为5.08和731,从数值上看总体高于健康香蕉植株处理,其中茎部差异显著;健康与发病‘巴西香牙蕉’植株内生细菌群落结构与组成差异显著,其中代尔夫特菌属(Delftia)、芽胞杆菌属(Bacillus)、海洋放线菌属(Salinispora)、不黏柄菌属(Asticcacaulis)和动胶菌属(Zoogloea)是抽蕾后健康‘巴西香牙蕉’植株的特有内生细菌属。[结论]‘巴西香牙蕉’植株对菌群吸收具有选择性,且根、茎、叶对微生物的选择性存在差异。‘巴西香牙蕉’内生细菌群落结构、多样性、丰富度与香蕉植株健康水平密切相关,病原菌入侵降低了香蕉植株对外源菌的选择性。
[Objectives]Fusarium wilt of banana is devastating banana plantations worldwide. In order to provide reference for biocontrol bacteria isolation and disease control,the endophytic bacterial communities in different parts of healthy and diseased banana( Musa AAA cavendish cv. Brazil) plants were studied. [Methods]The endophytic bacterial communities in different parts of healthy and diseased banana plants were profiled through MiSeq high-throughput sequencing of the bacterial 16 S r RNA amplicons.[Results]The endophytic bacterial community Shannon diversity and Chao1 richness index in roots of healthy banana plants were6.11 and 671,followed by leaves which were 5.49 and 370,and stems which were 2.50 and 237. The Shannon diversity and Chao1 richness index in root of diseased banana plants were 6.86 and 1 226,followed by stems which were 5.08 and 731,which were higher than those of healthy banana plants though the difference of Shannon diversity of root was insignificant. The endophytic bacterial community patterns were significantly different between the healthy and diseased banana plants. Delftia,Bacillus,Salinispora,Asticcacaulis and Zoogloea were indicator genera in healthy banana plants which were not detected in the diseased banana plants after buds. [Conclusions]The banana plants selectively absorb microbiome from the environment that different parts of banana plants had different endophytic community. The endophytic bacterial community composition and the diversity of banana plants were closely related to their health levels. The invasion of pathogen reduced the prevention level of banana plants to the microbial community outside of the plants.
[Objectives]Fusarium wilt of banana is devastating banana plantations worldwide. In order to provide reference for biocontrol bacteria isolation and disease control,the endophytic bacterial communities in different parts of healthy and diseased banana( Musa AAA cavendish cv. Brazil) plants were studied. [Methods]The endophytic bacterial communities in different parts of healthy and diseased banana plants were profiled through MiSeq high-throughput sequencing of the bacterial 16 S r RNA amplicons.[Results]The endophytic bacterial community Shannon diversity and Chao1 richness index in roots of healthy banana plants were6.11 and 671,followed by leaves which were 5.49 and 370,and stems which were 2.50 and 237. The Shannon diversity and Chao1 richness index in root of diseased banana plants were 6.86 and 1 226,followed by stems which were 5.08 and 731,which were higher than those of healthy banana plants though the difference of Shannon diversity of root was insignificant. The endophytic bacterial community patterns were significantly different between the healthy and diseased banana plants. Delftia,Bacillus,Salinispora,Asticcacaulis and Zoogloea were indicator genera in healthy banana plants which were not detected in the diseased banana plants after buds. [Conclusions]The banana plants selectively absorb microbiome from the environment that different parts of banana plants had different endophytic community. The endophytic bacterial community composition and the diversity of banana plants were closely related to their health levels. The invasion of pathogen reduced the prevention level of banana plants to the microbial community outside of the plants.
引文
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[10] De’Ath G. Multivariate regression trees:a new technique for modeling species-environment relationships[J]. Ecology,2002,83(4):1105-1117.
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[14]张苗苗.白木香中内生菌的分离鉴定及其分布规律的初步研究[D].广州:广州中医药大学,2017.Zhang M M. Apreliminary study on the isolation and identification of endophytic bacteria in white fragrant and distribution[D]. Guangzhou:Guangzhou University of Chinese Medicine,2017(in Chinese with English abstract).
[15] Hallmann J,Quadt-Hallmann A,Mahaffee W F,et al. Bacterial endophytes in agricultural crops[J]. Canadian Journal of Microbiology,1997,43(10):895-914.
[16]付业勤,蔡吉苗,刘先宝,等.香蕉内生细菌分离、活性评价及数量分布[J].热带作物学报,2007,28(4):78-83.Fu Y Q,Cai J M,Liu X B,et al. Isolation,bioactivity evaluation and quantity distribution of endophytic bacteria in banana[J]. Chinese Journal of Tropical Crops,2007,28(4):78-83(in Chinese with English abstract).
[17] Xue C,Penton C R,Shen Z,et al. Manipulating the banana rhizosphere microbiome for biological control of panama disease[J]. Scientific Reports,2015,5:11124.
[18] Snyder W C,Hansen H N. The species concept in fusarium[J]. American Journal of Botany,1940,27(2):64-67.
[19]蓝江林,刘波,焦会民,等.香蕉植株内生细菌群落多态性研究[J].热带亚热带植物学报,2012,20(3):285-291.Lan J L,Liu B,Jiao H M,et al. Community and diversity of endophytic bacteria in Musa nana[J]. Journal of Tropical and Subtropical Botany,2012,20(3):285-291(in Chinese with English abstract).
[20] Hardoim P R,van Overbeek L S,Elsas J D. Properties of bacterial endophytes and their proposed role in plant growth[J]. Trends in Microbiology,2008,16(10):463-471.
[21]李文英,彭智平,于俊红,等.香蕉根际促生菌的研究展望[J].热带作物学报,2011,32(1):182-187.Li W Y,Peng Z P,Yu J H,et al. Progress and prospect on plant growth-promoting rhizobacteria of banana rhizosphere[J]. Chinese Journal of Tropical Crops,2011,32(1):182-187(in Chinese with English abstract).
[22] Thomas P,Kumari S,Swarna G K,et al. Papaya shoot tip associated endophytic bacteria isolated from in vitro cultures and host-endophyte interaction in vitro and in vivo[J]. Canadian Journal of Microbiology,2007,53(3):380-390.
[23]孟宪法.滩涂植物菊芋根际固氮菌群落分析及其内生细菌作用机制[D].南京:南京农业大学,2015.Meng X F. Analysis on diazotrophic communities in the rhizosphere of beach plant Jerusalem artichoke and the mechanism of action of its endophytic bacteria[D]. Nanjing:Nanjing Agricultural University,2015(in Chinese with English abstract).
[24]栗方崎.海洋放线菌的分离筛选及其双组分信号转导系统功能的初步研究[D].济南:山东大学,2015.Li F Q. Isolation and screening of marine actinomycetes and study on the function of its two-component signal transduction system[D]. Jinan:Shandong University,2015(in Chinese with English abstract).
[25]李静,操一凡,丁佳兴,等.含复合菌群生物育苗基质的研制及其育苗效果[J].南京农业大学学报,2018,41(4):676-684. DOI:10.7685/jnau.201801009.Li J,Cao Y F,Ding J X,et al. Development of bio-nursery substrates containing PGPR flora and evaluation of their seedlings growth promoting effect[J]. Journal of Nanjing Agricultural University,2018,41(4):676-684(in Chinese with English abstract).
[26]彭爱琴.代尔夫特菌MTQ3拮抗相关基因tet R的克隆及功能验证[D].泰安:山东农业大学,2012.Peng A Q. Cloning and functional confirmation of the tet R related to antagonistic activity to Delftia tsuruhatensis MTQ3[D]. Tai’an:Shandong Agricultural University,2012(in Chinese with English abstract).
[27]郝晓娟.作物枯萎病生防细菌的研究[D].杭州:浙江大学,2006.Hao X J. Study on antagonistic bacteria for biocontrol of the Fusarium wilt[D]. Hangzhou:Zhejiang University,2006(in Chinese with English abstract).
[2] Butler D. Fungus threatens top banana[J]. Nature,2013,504(7479):195-196.
[3]赵娜,李荣,辛侃,等.茄科蔬菜轮作对高发枯萎病蕉园土壤可培养微生物的影响[J].热带作物学报,2014,35(8):1469-1474.Zhao N,Li R,Xin K,et al. Effects of different solanaceae crop rotations on the soil culturable microbes in an orchard with serious fusarium wilt disease[J]. Chinese Journal of Tropical Crops,2014,35(8):1469-1474(in Chinese with English abstract).
[4]朱小花,赵利敏,戴晓灵,等.香蕉枯萎病生物综合防控技术研究[J].广东农业科学,2013,40(7):86-88.Zhu X H,Zhao L M,Dai X L,et al. Study on biological comprehensive prevention and control technology of Fusarium wilt of banana[J]. Guangdong Agricultural Sciences,2013,40(7):86-88(in Chinese with English abstract).
[5]陈龙,梁子宁,朱华.植物内生菌研究进展[J].生物技术通报,2015,31(8):30-34.Chen L,Liang Z N,Zhu H. Research advances in the studies of plant entophytic[J]. Biotechnology Bulletin,2015,31(8):30-34(in Chinese with English abstract).
[6]陈博,朱军,孙前光,等.一株抗香蕉枯萎病内生细菌的分离鉴定及其抗病促生作用[J].微生物学通报,2011,38(2):199-205.Chen B,Zhu J,Sun Q G,et al. A bacterial endophyte from banana:its isolation,identification,activity to Fusarium wilt and PGPR effect to banana seedlings[J]. Microbiology China,2011,38(2):199-205(in Chinese with English abstract).
[7]王梦颖.香蕉内生菌多样性分析及广谱拮抗菌筛选[D].海口:海南大学,2014.Wang M Y. Diversity research of endophytes and broad-spectrum antagonistic bacterium screening from banana[D]. Haikou:Hainan University,2014(in Chinese with English abstract).
[8] Caporaso J G,Kuczynski J,Stombaugh J,et al. QIIME allows analysis of high-throughput community sequencing data[J]. Nature Methods,2010,7(5):335-336.
[9] Shepard R N. The analysis of proximities:multidimensional scaling with an unknown distance functionⅡ[J]. Psychometrika,1962,27(3):219-246.
[10] De’Ath G. Multivariate regression trees:a new technique for modeling species-environment relationships[J]. Ecology,2002,83(4):1105-1117.
[11] Clarke K R. Non-parametric multivariate analyses of changes in community structure[J]. Austral Ecology,1993,18(1):117-143.
[12] Anderson M J. A new method for non-parametric multivariate analysis of variance[J]. Austral Ecology,2001,26(1):32-46.
[13] Kirk J L,Beaudette L A,Hart M,et al. Methods of studying soil microbial diversity[J]. Journal of Microbiological Methods,2004,58(2):169-188.
[14]张苗苗.白木香中内生菌的分离鉴定及其分布规律的初步研究[D].广州:广州中医药大学,2017.Zhang M M. Apreliminary study on the isolation and identification of endophytic bacteria in white fragrant and distribution[D]. Guangzhou:Guangzhou University of Chinese Medicine,2017(in Chinese with English abstract).
[15] Hallmann J,Quadt-Hallmann A,Mahaffee W F,et al. Bacterial endophytes in agricultural crops[J]. Canadian Journal of Microbiology,1997,43(10):895-914.
[16]付业勤,蔡吉苗,刘先宝,等.香蕉内生细菌分离、活性评价及数量分布[J].热带作物学报,2007,28(4):78-83.Fu Y Q,Cai J M,Liu X B,et al. Isolation,bioactivity evaluation and quantity distribution of endophytic bacteria in banana[J]. Chinese Journal of Tropical Crops,2007,28(4):78-83(in Chinese with English abstract).
[17] Xue C,Penton C R,Shen Z,et al. Manipulating the banana rhizosphere microbiome for biological control of panama disease[J]. Scientific Reports,2015,5:11124.
[18] Snyder W C,Hansen H N. The species concept in fusarium[J]. American Journal of Botany,1940,27(2):64-67.
[19]蓝江林,刘波,焦会民,等.香蕉植株内生细菌群落多态性研究[J].热带亚热带植物学报,2012,20(3):285-291.Lan J L,Liu B,Jiao H M,et al. Community and diversity of endophytic bacteria in Musa nana[J]. Journal of Tropical and Subtropical Botany,2012,20(3):285-291(in Chinese with English abstract).
[20] Hardoim P R,van Overbeek L S,Elsas J D. Properties of bacterial endophytes and their proposed role in plant growth[J]. Trends in Microbiology,2008,16(10):463-471.
[21]李文英,彭智平,于俊红,等.香蕉根际促生菌的研究展望[J].热带作物学报,2011,32(1):182-187.Li W Y,Peng Z P,Yu J H,et al. Progress and prospect on plant growth-promoting rhizobacteria of banana rhizosphere[J]. Chinese Journal of Tropical Crops,2011,32(1):182-187(in Chinese with English abstract).
[22] Thomas P,Kumari S,Swarna G K,et al. Papaya shoot tip associated endophytic bacteria isolated from in vitro cultures and host-endophyte interaction in vitro and in vivo[J]. Canadian Journal of Microbiology,2007,53(3):380-390.
[23]孟宪法.滩涂植物菊芋根际固氮菌群落分析及其内生细菌作用机制[D].南京:南京农业大学,2015.Meng X F. Analysis on diazotrophic communities in the rhizosphere of beach plant Jerusalem artichoke and the mechanism of action of its endophytic bacteria[D]. Nanjing:Nanjing Agricultural University,2015(in Chinese with English abstract).
[24]栗方崎.海洋放线菌的分离筛选及其双组分信号转导系统功能的初步研究[D].济南:山东大学,2015.Li F Q. Isolation and screening of marine actinomycetes and study on the function of its two-component signal transduction system[D]. Jinan:Shandong University,2015(in Chinese with English abstract).
[25]李静,操一凡,丁佳兴,等.含复合菌群生物育苗基质的研制及其育苗效果[J].南京农业大学学报,2018,41(4):676-684. DOI:10.7685/jnau.201801009.Li J,Cao Y F,Ding J X,et al. Development of bio-nursery substrates containing PGPR flora and evaluation of their seedlings growth promoting effect[J]. Journal of Nanjing Agricultural University,2018,41(4):676-684(in Chinese with English abstract).
[26]彭爱琴.代尔夫特菌MTQ3拮抗相关基因tet R的克隆及功能验证[D].泰安:山东农业大学,2012.Peng A Q. Cloning and functional confirmation of the tet R related to antagonistic activity to Delftia tsuruhatensis MTQ3[D]. Tai’an:Shandong Agricultural University,2012(in Chinese with English abstract).
[27]郝晓娟.作物枯萎病生防细菌的研究[D].杭州:浙江大学,2006.Hao X J. Study on antagonistic bacteria for biocontrol of the Fusarium wilt[D]. Hangzhou:Zhejiang University,2006(in Chinese with English abstract).