内蒙古荒漠主要植物根际细菌菌群组成结构特征分析
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摘要
根际微生物作为土壤生态系统的生物组成部分,是陆地生态系统中重要的生命体,对所生存的微环境十分敏感。现在,土壤微生物指标已被公认是土壤生态系统变化的预警及敏感指标。
本文以内蒙古浑善达克沙地和毛乌素沙地主要植物根际土壤为研究对象,通过直接法提取沙蒿,锦鸡,黄柳等植物根际土壤的微生物总DNA,采用建立土壤细菌16S rDNA克隆文库的方法,对不同季节,不同沙地的不同植物根际微生物菌群进行比对研究,分析微生物群落结构的时空尺度变化,获取变化的作用因子,为研究和利用细菌与土壤环境间的相互作用提供理论依据,最终为治理土壤荒漠化提供理论依据。
研究结果表明:在植物生命活动不旺盛的11月,在植物根际土壤中,一般都是Pseudomonas为优势菌;到了4月和8月植物生命活动旺盛的时期,Rhizobiales和Sphingomonadales这两类与植物生长关系密切的菌得到了大量聚集,成为这两个时期的优势菌;研究还发现,同一地点不同植物的根际土壤中主要优势菌相同,非优势菌存在较大变化;不同地点的相同植物的根际土壤微生物主要优势菌基本相同,只是一些非优势菌存在变化。总之,植物根际的主要细菌类群与随季节而变化的植物的代谢活动有密切关系。
As biological components of ecosystems, rhizosphere soil bacteria are important organisms in terrestrial ecosystems, and they are very sensitive to micro-environment, their populations may be greatly affected by changes in environmental conditions. Now, soil bacteria has been recognized as an sensitive early warning indicator of soil ecosystem change.
This paper selected the rhizosphere soil of main plants in Hunshandake and Mu Us Sandland of Inner Mongolia as studying object, by direct extracted the total microorganism DNA of artemisia, caragana, yellow willow and other plants rhizosphere soil. Throμgh establishing the clone library of 16S rDNA of bacteria in soil, compared different seasons and different Desert’s different plants rhizosphere microflora, analyze bacterial community structural changes in spatial and temporal scales, to obtain the factors leading to the changes, for the study and use of The interaction between bacteria and the soil environment provide a theoretical basis, ultimately, for control of soil desertification, provide a theoretical basis.
The results showed that: In the plant life activities are not strong in November, in the rhizosphere soil, generally Pseudomonas is the dominant bacteria; to April and August, at this time plants have strong life activities, Rhizobiales and Sphingomonadales these two groups of bacteria to which the plant growth are chosely related gathered in these two periods became the most dominant of bacteria; the study also found that different species of plants rhizosphere soil bacteria of the same location, whose dominant bacteria are same, but there is a big changement in the non-dominant bacteria; the same plant rhizosphere bacteria at different locations, whose main advantaged bacteria is basically the same, but some non-dominant bacteria have some changements. In short, the main rhizosphere bacterial populations is closely related to metabolic activity of the plant which is changed with the seasons.
本文以内蒙古浑善达克沙地和毛乌素沙地主要植物根际土壤为研究对象,通过直接法提取沙蒿,锦鸡,黄柳等植物根际土壤的微生物总DNA,采用建立土壤细菌16S rDNA克隆文库的方法,对不同季节,不同沙地的不同植物根际微生物菌群进行比对研究,分析微生物群落结构的时空尺度变化,获取变化的作用因子,为研究和利用细菌与土壤环境间的相互作用提供理论依据,最终为治理土壤荒漠化提供理论依据。
研究结果表明:在植物生命活动不旺盛的11月,在植物根际土壤中,一般都是Pseudomonas为优势菌;到了4月和8月植物生命活动旺盛的时期,Rhizobiales和Sphingomonadales这两类与植物生长关系密切的菌得到了大量聚集,成为这两个时期的优势菌;研究还发现,同一地点不同植物的根际土壤中主要优势菌相同,非优势菌存在较大变化;不同地点的相同植物的根际土壤微生物主要优势菌基本相同,只是一些非优势菌存在变化。总之,植物根际的主要细菌类群与随季节而变化的植物的代谢活动有密切关系。
As biological components of ecosystems, rhizosphere soil bacteria are important organisms in terrestrial ecosystems, and they are very sensitive to micro-environment, their populations may be greatly affected by changes in environmental conditions. Now, soil bacteria has been recognized as an sensitive early warning indicator of soil ecosystem change.
This paper selected the rhizosphere soil of main plants in Hunshandake and Mu Us Sandland of Inner Mongolia as studying object, by direct extracted the total microorganism DNA of artemisia, caragana, yellow willow and other plants rhizosphere soil. Throμgh establishing the clone library of 16S rDNA of bacteria in soil, compared different seasons and different Desert’s different plants rhizosphere microflora, analyze bacterial community structural changes in spatial and temporal scales, to obtain the factors leading to the changes, for the study and use of The interaction between bacteria and the soil environment provide a theoretical basis, ultimately, for control of soil desertification, provide a theoretical basis.
The results showed that: In the plant life activities are not strong in November, in the rhizosphere soil, generally Pseudomonas is the dominant bacteria; to April and August, at this time plants have strong life activities, Rhizobiales and Sphingomonadales these two groups of bacteria to which the plant growth are chosely related gathered in these two periods became the most dominant of bacteria; the study also found that different species of plants rhizosphere soil bacteria of the same location, whose dominant bacteria are same, but there is a big changement in the non-dominant bacteria; the same plant rhizosphere bacteria at different locations, whose main advantaged bacteria is basically the same, but some non-dominant bacteria have some changements. In short, the main rhizosphere bacterial populations is closely related to metabolic activity of the plant which is changed with the seasons.
引文
1中国科学院南京土壤研究所微生物室,土壤微生物研究法[M].北京:科学出版社,1985
2刘占良,翟红,刘大群.植物根际的微生物互作及其在植物病害生物防治中的应用[J].河北农业大学学报,2003,26:183-186
3胡小加.根际微生物与植物营养[J].中国油料作物学报,1999,21(3):77-79
4吴建峰,林先贵.土壤微生物在促进植物生长方面的作用[J].土壤,2003,(l):18-21
5杨芳,徐秋芳.土壤微生物多样性研究进展[J].浙江林业科技,2002,22(6):39-42
6李延茂,胡江春,汪思龙等.森林生态系统中土壤微生物的作用与应用[J].应用生态学报,2004,15(10):1943-1946
7 DARCY L A. Study of soybean and lentil root exudates influence of soybean isofavonoids on the growth of rhizobia and some rhizospheric microorganisms[J]. Plant and Soil,1987,101:267-272
8朱丽霞,章家恩,刘文高.根系分泌物与根际微生物相互作用研究综述[J].生态化境.2003,12(1):103
9 MA R-X, FEN Y. Effect of allelopathic chemical on growth addenitrification of bacillus subtitis under anaerobic condition[J].Chem Ecol,1998,24:187-193
10刘洪升,宋秋华,李凤民.根分泌物对根际矿物营养及根际微生物的效应[J].西北植物学报,2002,22(3):693-702
11 GRAHAM R D.Genotypic differences in tolerance to manganese deficiency [M].In:Robson A.D,eds.Zinc in Soils and Plants.KluwerAcademic Publishers,Dordrecht,The Netherlands.1988:75-85.
12 Cecillia M.Joseph,Donald A.Phillips.Metabolites from soil bacteria affect plant water relation[J].Plant Physiology and Biochemistry,2003,41:189-192
13 Schippers B,Bakker AW,Backker PA.Interactions of deleterious and beneficial rhizosphere microoganisms[J].Annu Rev Phytopathol,1987,25:339-358
14 Bending GD,Lincoln SD.Walker A. In-field spatial variability in the degradation of the phenylurea herbicide isoproturon is the resule of interactions between degradative Sphingomonas spp. And soil pH[J].Appl Environ Microiol,2003,69:827-834
15 Takeuchi M,Sakane T.Yokota. Taxonomic study of bacteria isolated from plants: proposal of Sphingomonas rosasp.nov., Sphingomonas pruni sp.Nov., Sphingomonas asaccharolytica sp.nov,.and Sphingomonas mali sp. Nov.[J].IntJ Syst Bacterilo,1995,141:99-100
16 Berg G,Ballin G.Bacterial antagonists to Verticillium dahliae[J]Kleb.J Phytopatho,1994,141:99-100
17胡杰,何晓红等.鞘氨醇单胞菌研究进展[J].应用与环境微生物学报,2007,13(3):431-437
18 Saygili H. et al . Laboratory and field t rials with selected microorganisms as biocontrol agents for fire blight[J].Acta Horticulturae,1999,5:655-661
19 SARWARM,KREMER RJ.Determination of Bacterially Derived Auxins Using a microplate Method[J].Lett Appl Microbiol,1995,20:282-285
20 Kim ,K. Y. et al . Solubilization of hydroxyappatite by Enterobacter agglomerans and cloned Escherichia coli in culture medium[J].Biol.Fertil. Soils,1997,24:347-352
21沈德龙,冯永君.成团肠杆菌的生物功能多样性及其分类最新进展微生物学杂志[J]. 2002,22(1):40-42
22程桂荪.根瘤菌和内生菌根混合接种对三叶草增产的效应[J].土壤,1986,18(3):136
23 Gardner C.MyeorrhizaeofactiollrllizalPlants[J].Mierobiology and Biotechnology,1986,2(l):147-160
24 Van Der Heijden,et a1.Different arbuscular mycorrhizal fungal species are potential determinants ofplant community structure[J].Ecology,1998,79(6):2082-2091
25杨海君,谭周进等.假单胞菌的生物防治作用研究[J].中国生态农业学报.2006,12(3):160
26 Hartmann A,Schmid M,Wenzel W,Hinsinger Ph.International Congress Rhizosphere Perspectives and Challenges-A Tribute to Lorenz Hiltner[J].Research Center for Environment and Health,2005,1:3-6
27刘国奇,蒋如璋.韭菜根际荧光假单胞菌株的分离和初步研究[J].微生物学通报,1999,26(3):189-191
28 Maire N,Lackzo E,et al.Organic matter cycling in grassland soils of the Swiss Jura mountains: biodiversity and strategies of the living communities[J].Soil Biology and Bioehemistry.1999,31:1281-1293
29 Bentivega S P, Morton J B. Mycol Res,Microbiota do Solo e Qualidade Ambiental[J] Microbiol.Methods.1994,98:1419-1426
30 Gruntzig V,Stres B,Ayala HL et al.Improved Protocol for T-RFLP by Capillary Electrophoresis[E].2002:516-518
31 Ian C. Anderson,John W. G. Cairney. Diversity and ecology of soil fungal communities:increased understanding through the application of molecular techniques.[J].Environmental Microbiology 2004,6(8):769-779
32 Moeseneder MM, Winter C, Arrieta JM, Herndl GJ. Terminal-restriction fragment length polymorphism (T-RFLP) screening of a marine archaeal clone library to determine the different phylotypes.[J]Microbiol.Methods,2001,44:159-172
33 Muyzer G, De Waal EC, Uitterlinden AG. Profiling of complex microbial populations by denaturing gradient gel electrophoresis analysis of polymerase chain reaction-amplified genes coding for 16S rRNA[J].Appl.Environmental.Microbiol,1993, 59:695-700
34 Zhou J Z,Xia B C,Huang H S,et al.Microbial diversity and heterogeneity in sandy subsurface soils[J].Applied and Environmental Microbiology,2004,70:1723-1734
35 Muyzer G. DGGE/TGGE a method for identifying genes from natural ecosystems[J].Curr Opin Microbiol,1999,2(3):317-322
36王天龙.内蒙古鄂尔多斯几种典型固沙植物根际微生物研究[D].四川农业大学:2007
37 Karlen D L,Gardner J C,Rosek M J.A Soil quality framework for evaluating the impact of CRP[J].Journal of Production Agriculture.1998,11(1):56-60
38 Stenberg B.Monitoring soil quality of arable land:microbiological indicators[J].Acta Agriculturae Scandinavica,Section B,Soil and Plant Sciencem1999,49:1-24
39赵翔,陈绍兴,谢志雄,等.高产铁载体荧光假单胞菌Pseudomonas fluorescens sp-2f的筛选鉴定及其铁载体特性研究[J].微生物学报,2006,46(5):691-695
40胡江春,薛德林,马成新,王书锦.植物根际促生菌(PGPR)的研究与应用前景[J].应用生态学报,2004,15(10):1963
41吴凡等.桑树根际微生物研究[D].山东农业大学:2007
42 Filion M, Hamelin R C, Benier L, et al.Molecular profiling of rhizosphere microbial communities associated with healthy and diseased black spruce (Picea mariana) seedlings grown in a nursery[J].Appl Environ Microbiol,2004,70(6): 3541-3551
43祖元刚,高崇洋.喜树替代紫茎泽兰过程中根际微生物群落特征[J].生命科学,2006,36(5):459-467
44 Miethling R,Wieland G.,Backhaus H,et al.Variation of microbial rhizosphere communities in response to crop spe-cies, soil origin, and inoculation withSinorhizobium meliloti L
33[J].Microbial Ecology,2000,40:43-56
45 P.Marschner,C.-H.Yang,R.Lieberei,et al.Soil and plant specific effects on bacterial community composition inthe rhizosphere[J],Soil Biology and Biochemistry, 2001,33:1437-1445
46赵庆节,梁丹涛等.五种人工湿地植物根际细菌多样性的研究[J].三峡环境与生态.2008,1(3):11-13
2刘占良,翟红,刘大群.植物根际的微生物互作及其在植物病害生物防治中的应用[J].河北农业大学学报,2003,26:183-186
3胡小加.根际微生物与植物营养[J].中国油料作物学报,1999,21(3):77-79
4吴建峰,林先贵.土壤微生物在促进植物生长方面的作用[J].土壤,2003,(l):18-21
5杨芳,徐秋芳.土壤微生物多样性研究进展[J].浙江林业科技,2002,22(6):39-42
6李延茂,胡江春,汪思龙等.森林生态系统中土壤微生物的作用与应用[J].应用生态学报,2004,15(10):1943-1946
7 DARCY L A. Study of soybean and lentil root exudates influence of soybean isofavonoids on the growth of rhizobia and some rhizospheric microorganisms[J]. Plant and Soil,1987,101:267-272
8朱丽霞,章家恩,刘文高.根系分泌物与根际微生物相互作用研究综述[J].生态化境.2003,12(1):103
9 MA R-X, FEN Y. Effect of allelopathic chemical on growth addenitrification of bacillus subtitis under anaerobic condition[J].Chem Ecol,1998,24:187-193
10刘洪升,宋秋华,李凤民.根分泌物对根际矿物营养及根际微生物的效应[J].西北植物学报,2002,22(3):693-702
11 GRAHAM R D.Genotypic differences in tolerance to manganese deficiency [M].In:Robson A.D,eds.Zinc in Soils and Plants.KluwerAcademic Publishers,Dordrecht,The Netherlands.1988:75-85.
12 Cecillia M.Joseph,Donald A.Phillips.Metabolites from soil bacteria affect plant water relation[J].Plant Physiology and Biochemistry,2003,41:189-192
13 Schippers B,Bakker AW,Backker PA.Interactions of deleterious and beneficial rhizosphere microoganisms[J].Annu Rev Phytopathol,1987,25:339-358
14 Bending GD,Lincoln SD.Walker A. In-field spatial variability in the degradation of the phenylurea herbicide isoproturon is the resule of interactions between degradative Sphingomonas spp. And soil pH[J].Appl Environ Microiol,2003,69:827-834
15 Takeuchi M,Sakane T.Yokota. Taxonomic study of bacteria isolated from plants: proposal of Sphingomonas rosasp.nov., Sphingomonas pruni sp.Nov., Sphingomonas asaccharolytica sp.nov,.and Sphingomonas mali sp. Nov.[J].IntJ Syst Bacterilo,1995,141:99-100
16 Berg G,Ballin G.Bacterial antagonists to Verticillium dahliae[J]Kleb.J Phytopatho,1994,141:99-100
17胡杰,何晓红等.鞘氨醇单胞菌研究进展[J].应用与环境微生物学报,2007,13(3):431-437
18 Saygili H. et al . Laboratory and field t rials with selected microorganisms as biocontrol agents for fire blight[J].Acta Horticulturae,1999,5:655-661
19 SARWARM,KREMER RJ.Determination of Bacterially Derived Auxins Using a microplate Method[J].Lett Appl Microbiol,1995,20:282-285
20 Kim ,K. Y. et al . Solubilization of hydroxyappatite by Enterobacter agglomerans and cloned Escherichia coli in culture medium[J].Biol.Fertil. Soils,1997,24:347-352
21沈德龙,冯永君.成团肠杆菌的生物功能多样性及其分类最新进展微生物学杂志[J]. 2002,22(1):40-42
22程桂荪.根瘤菌和内生菌根混合接种对三叶草增产的效应[J].土壤,1986,18(3):136
23 Gardner C.MyeorrhizaeofactiollrllizalPlants[J].Mierobiology and Biotechnology,1986,2(l):147-160
24 Van Der Heijden,et a1.Different arbuscular mycorrhizal fungal species are potential determinants ofplant community structure[J].Ecology,1998,79(6):2082-2091
25杨海君,谭周进等.假单胞菌的生物防治作用研究[J].中国生态农业学报.2006,12(3):160
26 Hartmann A,Schmid M,Wenzel W,Hinsinger Ph.International Congress Rhizosphere Perspectives and Challenges-A Tribute to Lorenz Hiltner[J].Research Center for Environment and Health,2005,1:3-6
27刘国奇,蒋如璋.韭菜根际荧光假单胞菌株的分离和初步研究[J].微生物学通报,1999,26(3):189-191
28 Maire N,Lackzo E,et al.Organic matter cycling in grassland soils of the Swiss Jura mountains: biodiversity and strategies of the living communities[J].Soil Biology and Bioehemistry.1999,31:1281-1293
29 Bentivega S P, Morton J B. Mycol Res,Microbiota do Solo e Qualidade Ambiental[J] Microbiol.Methods.1994,98:1419-1426
30 Gruntzig V,Stres B,Ayala HL et al.Improved Protocol for T-RFLP by Capillary Electrophoresis[E].2002:516-518
31 Ian C. Anderson,John W. G. Cairney. Diversity and ecology of soil fungal communities:increased understanding through the application of molecular techniques.[J].Environmental Microbiology 2004,6(8):769-779
32 Moeseneder MM, Winter C, Arrieta JM, Herndl GJ. Terminal-restriction fragment length polymorphism (T-RFLP) screening of a marine archaeal clone library to determine the different phylotypes.[J]Microbiol.Methods,2001,44:159-172
33 Muyzer G, De Waal EC, Uitterlinden AG. Profiling of complex microbial populations by denaturing gradient gel electrophoresis analysis of polymerase chain reaction-amplified genes coding for 16S rRNA[J].Appl.Environmental.Microbiol,1993, 59:695-700
34 Zhou J Z,Xia B C,Huang H S,et al.Microbial diversity and heterogeneity in sandy subsurface soils[J].Applied and Environmental Microbiology,2004,70:1723-1734
35 Muyzer G. DGGE/TGGE a method for identifying genes from natural ecosystems[J].Curr Opin Microbiol,1999,2(3):317-322
36王天龙.内蒙古鄂尔多斯几种典型固沙植物根际微生物研究[D].四川农业大学:2007
37 Karlen D L,Gardner J C,Rosek M J.A Soil quality framework for evaluating the impact of CRP[J].Journal of Production Agriculture.1998,11(1):56-60
38 Stenberg B.Monitoring soil quality of arable land:microbiological indicators[J].Acta Agriculturae Scandinavica,Section B,Soil and Plant Sciencem1999,49:1-24
39赵翔,陈绍兴,谢志雄,等.高产铁载体荧光假单胞菌Pseudomonas fluorescens sp-2f的筛选鉴定及其铁载体特性研究[J].微生物学报,2006,46(5):691-695
40胡江春,薛德林,马成新,王书锦.植物根际促生菌(PGPR)的研究与应用前景[J].应用生态学报,2004,15(10):1963
41吴凡等.桑树根际微生物研究[D].山东农业大学:2007
42 Filion M, Hamelin R C, Benier L, et al.Molecular profiling of rhizosphere microbial communities associated with healthy and diseased black spruce (Picea mariana) seedlings grown in a nursery[J].Appl Environ Microbiol,2004,70(6): 3541-3551
43祖元刚,高崇洋.喜树替代紫茎泽兰过程中根际微生物群落特征[J].生命科学,2006,36(5):459-467
44 Miethling R,Wieland G.,Backhaus H,et al.Variation of microbial rhizosphere communities in response to crop spe-cies, soil origin, and inoculation withSinorhizobium meliloti L
33[J].Microbial Ecology,2000,40:43-56
45 P.Marschner,C.-H.Yang,R.Lieberei,et al.Soil and plant specific effects on bacterial community composition inthe rhizosphere[J],Soil Biology and Biochemistry, 2001,33:1437-1445
46赵庆节,梁丹涛等.五种人工湿地植物根际细菌多样性的研究[J].三峡环境与生态.2008,1(3):11-13