转MdSOS2L1基因苹果植株根际微生物多样性的研究
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摘要
以野生型‘嘎拉’苹果和转MdSOS2L1基因植株为试材,利用高通量测序技术研究转MdSOS2L1苹果植株对根际微生物的影响。结果表明,转基因与野生型植株根际微生物群落丰富度和多样性存在较大差异,其中原核微生物群落变化最为明显。转MdSOS2L1植株根际酸杆菌门丰度增加较多,蓝藻、放线菌门丰度略有下降。同时,转基因植株根际土壤pH值低于野生型,并且转基因植株根际土壤的苹果酸、柠檬酸、草酸含量明显升高,两者非根际土无明显差异,转基因植株根际微生物的差异极有可能与MdSOS2L1基因介导的有机酸分泌有关。
Rhizosphere microorganisms have great effects on plant growth and development. The exudates of plants can also affect the distribution and species of microorganisms in rhizosphere. In this study,a high throughput sequencing technology was used to clarify the effects of MdSOS2 L1 transgenic Gala apple plants on rhizosphere microorganisms. The results showed that there were great differences in the abundance and diversity of rhizosphere microorganisms among the MdSOS2 L1 transgenic apple plants in compared with wild type apple plants. Among them,the rhizosphere of prokaryote community varied obviously in phylum level. The abundance of Acidobacteria in transgenic apple rhizosphere was higher than that in wild type apple plants. The abundance of Cyanobacteria and Actinobacteria was lower in transgenic apple rhizosphere microorganisms. Meanwhile,the pH value of rhizosphere soil of transgenic plants was lower than that of wild type plants. And the contents of oxalic acid,citric acid and L-malate in transgenic plants root were higher than that in wild type plants rhizosphere soil,but no significant differences occurred among the bulk soil between these two types of apple plants. Above of all,it is suggested that the MdSOS2 L1-mediated secretion of organic acids contributes to the difference in rhizosphere microorganisms.
Rhizosphere microorganisms have great effects on plant growth and development. The exudates of plants can also affect the distribution and species of microorganisms in rhizosphere. In this study,a high throughput sequencing technology was used to clarify the effects of MdSOS2 L1 transgenic Gala apple plants on rhizosphere microorganisms. The results showed that there were great differences in the abundance and diversity of rhizosphere microorganisms among the MdSOS2 L1 transgenic apple plants in compared with wild type apple plants. Among them,the rhizosphere of prokaryote community varied obviously in phylum level. The abundance of Acidobacteria in transgenic apple rhizosphere was higher than that in wild type apple plants. The abundance of Cyanobacteria and Actinobacteria was lower in transgenic apple rhizosphere microorganisms. Meanwhile,the pH value of rhizosphere soil of transgenic plants was lower than that of wild type plants. And the contents of oxalic acid,citric acid and L-malate in transgenic plants root were higher than that in wild type plants rhizosphere soil,but no significant differences occurred among the bulk soil between these two types of apple plants. Above of all,it is suggested that the MdSOS2 L1-mediated secretion of organic acids contributes to the difference in rhizosphere microorganisms.
引文
Allard S M,Walsh C S,Wallis A E,Ottesen A R,Brown E W,Micallef S A.2016.Solanum lycopersicum(tomato)hosts robust phyllosphere and rhizosphere bacterial communities when grown in soil amended with various organic and synthetic fertilizers.Science of the Total Environment,573:555-563.
Bates S T,Clemente J C,Flores G E,Walters W A,Parfrey L W,Knight R,Fierer N.2013.Global biogeography of highly diverse protistan communities in soil.Isme Journal,7(3):652-659.
Bednarek P,Kwon C,Schulze-Lefert P.2010.Not a peripheral issue:secretion in plant-microbe interactions.Current Opinion in Plant Biology,13(4):378-387.
Cheng N H,Pittman J K,Zhu J K,Hirschi K D.2004.The protein kinase SOS2 activates the Arabidopsis H+/Ca2+antiporter CAX1 to integrate calcium transport and salt tolerance.Journal of Biological Chemistry,279(4):2922-2926.
De-La-Pe?a C,Lei Z,Watson B S,Sumner L W,Vivanco J M.2008.Root-microbe communication through protein secretion.Journal of Biological Chemistry,283(37):25247.
Donegan K K,Palm C J,Fieland V J,Porteous L A,Ganio L M,Schaller D L.1995.Changes in levels,species and DNA fingerprints of soil microorganisms associated with cotton expressing the Bacillus thuringiensis var.kurstaki,endotoxin.Applied Soil Ecology,2(2):111-124.
Hu D G,Ma Q J,Sun C H,Sun M H,You C X,Hao Y J.2015.Overexpression of Md SOS2L1,a CIPK protein kinase,increases the antioxidant metabolites to enhance salt tolerance in apple and tomato.Physiologia Plantarum,156(2):201-214.
Hückelhoven R.2007.Transport and secretion in plant-microbe interactions.Current Opinion in Plant Biology,10(6):573.
Li Qiang,Wen Huancheng,Hu Cairong.2007.Difference between international and domestic methods in determining soil p H.Soils,39(3):488-491.(in Chinese)李强,文唤成,胡彩荣.2007.土壤p H值的测定国际国内方法差异研究.土壤,39(3):488-491.
Liu J,Ishitani M,Halfter U,Kim C S,Zhu J K.2000.The Arabidopsis thaliana SOS2 gene encodes a protein kinase that is required for salt tolerance.Proceedings of the National Academy of Sciences,97(7):3730-3734.
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Luo Yongqing,Zhao Xueyong,Li Meixia.2012.Ecological effect of plant root exudates and related affecting factors.Chinese Journal of Applied Ecology,23(12):3496-3504.(in Chinese)罗永清,赵学勇,李美霞.2012.植物根系分泌物生态效应及其影响因素研究综述.应用生态学报,23(12):3496-3504.
Peng Q A,Shaaban M,Wu Y,Hu R,Wang B,Wang J.2016.The diversity of iron reducing bacteria communities in subtropical paddy soils of China.Applied Soil Ecology,101(4):20-27.
Qin Nan,Li Dongfang,Yang Ruifu.2011.Next-generation sequencing technologies and the application in microbiology-a review.Acta Microbiologica Sinica,51(4):445-457.(in Chinese)秦楠,栗东芳,杨瑞馥.2011.高通量测序技术及其在微生物学研究中的应用.微生物学报,51(4):445-457.
Qiu Quan-sheng,Guo Yan,Margaret A,Dietrich,Karen S,Schumaker,Zhu Jian-kang.2002.Regulation of SOS1,a plasma membrane Na+/H+exchanger in Arabidopsis thaliana,by SOS2 and SOS3.Proceedings of the National Academy of Sciences of the United States of America,99(12):8436.
Quan R,Wang J,Yang D,Zhang H,Zhang Z,Huang R.2017.Ein3 and SOS2 synergistically modulate plant salt tolerance.Sci Rep,7:44637.
Rosier A,Bishnoi U,Lakshmanan V,Sherrier D J,Bais H P.2016.A perspective on inter-kingdom signaling in plant-beneficial microbe interactions.Plant Molecular Biology,90(6):537.
Shi J Y,Yuan X F,Lin H R,Yang Y Q,Li Z Y.2011.Differences in soil properties and bacterial communities between the rhizosphere and bulk soil and among different production areas of the medicinal plant Fritillaria thunbergii.International Journal of Molecular Sciences,12(6):3770-3785.
Sun Baoli,Chi Jie,Fan Zhongnan,He Xiaowei,Li Yanhua,Tong Chengfeng.2010.Determination of organic acids from integrated system of soil and plant.Environmental Science&Technology,33(9):137-141.(in Chinese)孙宝利,赤杰,范中南,贺小蔚,李艳华,仝乘风.2010.土壤及植物复合体系中有机酸的测定.环境科学与技术,33(9):137-141.
Verbon E H,Liberman L M.2016.Beneficial microbes affect endogenous mechanisms controlling root development.Trends in Plant Science,21(3):218.
Wang Xingchun,Yang Zhirong,Wang Min,Li Wei,Li Shengcai.2012.High-throughput sequencing technology and its application.China Biotechnology,32(1):109-114.(in Chinese)王兴春,杨致荣,王敏,李玮,李生才.2012.高通量测序技术及其应用.中国生物工程杂志,32(1):109-114.
Wartrud L S,Seidler R J,Huang P M,Adriano D C,Logan T J,Checkai R T.1998.Nontarget ecological effects of plant,microbial,and chemical introductions to terrestrial systems//Soil Chemistry and Ecosystem Health.Wisonsin:Soil Science Society of America:313-340.
Wu Linkun,Lin Xiangmin,Lin Wenxiong.2014.Advances and perspective in research on plant-soil-microbe interactions mediated by root exudates.Chinese Journal of Plant Ecology,38(3):298-310.(in Chinese)吴林坤,林向民,林文雄.2014.根系分泌物介导下植物-土壤-微生物互作关系研究进展与展望.植物生态学报,38(3):298-310.
Xu W,Wang Z,Wu F.2015.The effect of D123 wheat as a companion crop on soil enzyme activities,microbial biomass and microbial communities in the rhizosphere of watermelon.Frontiers in Microbiology,6:899.
Yin Chengmiao,Wang Mei,Wang Jiayan,Chen Xuesen,Shen Xiang,Zhang Min,Mao Zhiquan.2017.The research advance on apple replant disease.Acta Horticulturae Sinica,44(11):2215-2230.(in Chinese)尹承苗,王玫,王嘉艳,陈学森,沈向,张民,毛志泉.2017.苹果连作障碍研究进展.园艺学报,44(11):2215-2230.
Yuan H X,Zhang J Z,Guo J F,Xu X P,Li Y R.2005.Activities of microbes and enzymes in soil after growing transgenic rice with two extra anti-fungus genes.Acta Pedologica Sinica,42(1):122-126.
Zhan Fangdong.2004.Community structures and dynamics of microorganisms in the rhizosphere of flue-cured tobacco[M.D.Dissertation].Chongqing:Agricultural University of Southwest.(in Chinese)湛方栋.2004.烤烟根际微生物的群落结构及其动态变化的研究[硕士论文].重庆:西南农业大学.
Zhang Quanfang,Li Jun,Fan Zhongxue,Yang Lianqun,Bu Xun.2013.Application of high-throughput sequencing technology in agricultural research.Shandong Agricultural Sciences,41(1):137-140.(in Chinese)张全芳,李军,范仲学,杨连群,步迅.2013.高通量测序技术在农业研究中的应用.山东农业科学,41(1):137-140.
Zhou Deqing.2011.Essential Microbiology.3rd ed.Beijing:Higher Education Press.(in Chinese)周德庆.2011.微生物学教程.3版.北京:高等教育出版社.
Zhu J K.2002.Salt and drought stress signal transduction in plants.Annual Review of Plant Biology,53(53):247.
Bates S T,Clemente J C,Flores G E,Walters W A,Parfrey L W,Knight R,Fierer N.2013.Global biogeography of highly diverse protistan communities in soil.Isme Journal,7(3):652-659.
Bednarek P,Kwon C,Schulze-Lefert P.2010.Not a peripheral issue:secretion in plant-microbe interactions.Current Opinion in Plant Biology,13(4):378-387.
Cheng N H,Pittman J K,Zhu J K,Hirschi K D.2004.The protein kinase SOS2 activates the Arabidopsis H+/Ca2+antiporter CAX1 to integrate calcium transport and salt tolerance.Journal of Biological Chemistry,279(4):2922-2926.
De-La-Pe?a C,Lei Z,Watson B S,Sumner L W,Vivanco J M.2008.Root-microbe communication through protein secretion.Journal of Biological Chemistry,283(37):25247.
Donegan K K,Palm C J,Fieland V J,Porteous L A,Ganio L M,Schaller D L.1995.Changes in levels,species and DNA fingerprints of soil microorganisms associated with cotton expressing the Bacillus thuringiensis var.kurstaki,endotoxin.Applied Soil Ecology,2(2):111-124.
Hu D G,Ma Q J,Sun C H,Sun M H,You C X,Hao Y J.2015.Overexpression of Md SOS2L1,a CIPK protein kinase,increases the antioxidant metabolites to enhance salt tolerance in apple and tomato.Physiologia Plantarum,156(2):201-214.
Hückelhoven R.2007.Transport and secretion in plant-microbe interactions.Current Opinion in Plant Biology,10(6):573.
Li Qiang,Wen Huancheng,Hu Cairong.2007.Difference between international and domestic methods in determining soil p H.Soils,39(3):488-491.(in Chinese)李强,文唤成,胡彩荣.2007.土壤p H值的测定国际国内方法差异研究.土壤,39(3):488-491.
Liu J,Ishitani M,Halfter U,Kim C S,Zhu J K.2000.The Arabidopsis thaliana SOS2 gene encodes a protein kinase that is required for salt tolerance.Proceedings of the National Academy of Sciences,97(7):3730-3734.
Lottmann J,Heuer H,Vries J.2000.Establishment of introduced antagonistic bacteria in the rhizosphere of transgenic potatoes and their effect on the bacterial community.FEMS Microbial Ecol,33:41-49.
Luo Yongqing,Zhao Xueyong,Li Meixia.2012.Ecological effect of plant root exudates and related affecting factors.Chinese Journal of Applied Ecology,23(12):3496-3504.(in Chinese)罗永清,赵学勇,李美霞.2012.植物根系分泌物生态效应及其影响因素研究综述.应用生态学报,23(12):3496-3504.
Peng Q A,Shaaban M,Wu Y,Hu R,Wang B,Wang J.2016.The diversity of iron reducing bacteria communities in subtropical paddy soils of China.Applied Soil Ecology,101(4):20-27.
Qin Nan,Li Dongfang,Yang Ruifu.2011.Next-generation sequencing technologies and the application in microbiology-a review.Acta Microbiologica Sinica,51(4):445-457.(in Chinese)秦楠,栗东芳,杨瑞馥.2011.高通量测序技术及其在微生物学研究中的应用.微生物学报,51(4):445-457.
Qiu Quan-sheng,Guo Yan,Margaret A,Dietrich,Karen S,Schumaker,Zhu Jian-kang.2002.Regulation of SOS1,a plasma membrane Na+/H+exchanger in Arabidopsis thaliana,by SOS2 and SOS3.Proceedings of the National Academy of Sciences of the United States of America,99(12):8436.
Quan R,Wang J,Yang D,Zhang H,Zhang Z,Huang R.2017.Ein3 and SOS2 synergistically modulate plant salt tolerance.Sci Rep,7:44637.
Rosier A,Bishnoi U,Lakshmanan V,Sherrier D J,Bais H P.2016.A perspective on inter-kingdom signaling in plant-beneficial microbe interactions.Plant Molecular Biology,90(6):537.
Shi J Y,Yuan X F,Lin H R,Yang Y Q,Li Z Y.2011.Differences in soil properties and bacterial communities between the rhizosphere and bulk soil and among different production areas of the medicinal plant Fritillaria thunbergii.International Journal of Molecular Sciences,12(6):3770-3785.
Sun Baoli,Chi Jie,Fan Zhongnan,He Xiaowei,Li Yanhua,Tong Chengfeng.2010.Determination of organic acids from integrated system of soil and plant.Environmental Science&Technology,33(9):137-141.(in Chinese)孙宝利,赤杰,范中南,贺小蔚,李艳华,仝乘风.2010.土壤及植物复合体系中有机酸的测定.环境科学与技术,33(9):137-141.
Verbon E H,Liberman L M.2016.Beneficial microbes affect endogenous mechanisms controlling root development.Trends in Plant Science,21(3):218.
Wang Xingchun,Yang Zhirong,Wang Min,Li Wei,Li Shengcai.2012.High-throughput sequencing technology and its application.China Biotechnology,32(1):109-114.(in Chinese)王兴春,杨致荣,王敏,李玮,李生才.2012.高通量测序技术及其应用.中国生物工程杂志,32(1):109-114.
Wartrud L S,Seidler R J,Huang P M,Adriano D C,Logan T J,Checkai R T.1998.Nontarget ecological effects of plant,microbial,and chemical introductions to terrestrial systems//Soil Chemistry and Ecosystem Health.Wisonsin:Soil Science Society of America:313-340.
Wu Linkun,Lin Xiangmin,Lin Wenxiong.2014.Advances and perspective in research on plant-soil-microbe interactions mediated by root exudates.Chinese Journal of Plant Ecology,38(3):298-310.(in Chinese)吴林坤,林向民,林文雄.2014.根系分泌物介导下植物-土壤-微生物互作关系研究进展与展望.植物生态学报,38(3):298-310.
Xu W,Wang Z,Wu F.2015.The effect of D123 wheat as a companion crop on soil enzyme activities,microbial biomass and microbial communities in the rhizosphere of watermelon.Frontiers in Microbiology,6:899.
Yin Chengmiao,Wang Mei,Wang Jiayan,Chen Xuesen,Shen Xiang,Zhang Min,Mao Zhiquan.2017.The research advance on apple replant disease.Acta Horticulturae Sinica,44(11):2215-2230.(in Chinese)尹承苗,王玫,王嘉艳,陈学森,沈向,张民,毛志泉.2017.苹果连作障碍研究进展.园艺学报,44(11):2215-2230.
Yuan H X,Zhang J Z,Guo J F,Xu X P,Li Y R.2005.Activities of microbes and enzymes in soil after growing transgenic rice with two extra anti-fungus genes.Acta Pedologica Sinica,42(1):122-126.
Zhan Fangdong.2004.Community structures and dynamics of microorganisms in the rhizosphere of flue-cured tobacco[M.D.Dissertation].Chongqing:Agricultural University of Southwest.(in Chinese)湛方栋.2004.烤烟根际微生物的群落结构及其动态变化的研究[硕士论文].重庆:西南农业大学.
Zhang Quanfang,Li Jun,Fan Zhongxue,Yang Lianqun,Bu Xun.2013.Application of high-throughput sequencing technology in agricultural research.Shandong Agricultural Sciences,41(1):137-140.(in Chinese)张全芳,李军,范仲学,杨连群,步迅.2013.高通量测序技术在农业研究中的应用.山东农业科学,41(1):137-140.
Zhou Deqing.2011.Essential Microbiology.3rd ed.Beijing:Higher Education Press.(in Chinese)周德庆.2011.微生物学教程.3版.北京:高等教育出版社.
Zhu J K.2002.Salt and drought stress signal transduction in plants.Annual Review of Plant Biology,53(53):247.