羊蹄根际土壤微生物研究
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摘要
羊蹄(Rumex patientia)为蓼科第二大属酸模属植物，全世界约有150种。我国酸模属植物资源极为丰富，有26种，分布全国各省区。羊蹄药用始载于《神农本草经》。该类中草药在我国民间以止血和治疗疥癣著称。现代药理学研究表明其具有抗真菌、抗肿瘤、抗病毒和抗氧化等作用。植物的化学分析显示，羊蹄具有丰富的蒽醌，单宁酸，萘衍生物等。
本论文首先采用微生物培养及非培养的方法研究吉林市4个生境羊蹄根际与非根际土壤微生物多样性；其次采用微生物培养的方法研究长春市2个生境羊蹄根际与非根际土壤微生物；最后对吉林市与长春市相似生境的羊蹄根际土壤微生物进行了比较分析。
主要结果如下：
1.采用微生物培养的方法研究吉林市4个生境羊蹄根际与非根际土壤微生物主要由细菌、放线菌组成，真菌最少。根际与非根际土壤相比，微生物群落更多。
(1)主成分分析显示自生固氮菌、氨化细菌是根际土壤的最主要影响因子，另外，总细菌和放线菌也是重要的影响因子。硫化细菌，总细菌和氨化细菌是非根际土壤的最主要影响因子，另外，放线菌和真菌也是重要的影响因子。
(2)皮尔森相关系数分析显示吉林市4个生境羊蹄根际土壤放线菌的丰富度与湿度及腐殖质呈显著负相关，氨化细菌的丰富度与总P含量呈显著负相关，自生固氮菌的丰富度与总P含量呈非常显著负相关。在非根际土壤，总细菌的丰富度与总K含量呈显著负相关，放线菌的丰富度与pH呈显著正相关，自生固氮菌的丰富度与pH呈显著负相关。
2.长春市2个生境羊蹄根际土壤微生物群落数量多于非根际土壤。
3.吉林市与长春市两个相似生境的羊蹄根际土壤微生物比较分析结果表明，近水生境、向阳生境羊蹄根际土壤微生物数量吉林市均高于长春市。
4.采用16SrDNA技术研究吉林市4个不同生境下羊蹄根际与非根际土壤特有的克隆多于共有的克隆；并且根际土壤特有的克隆多于非根际土壤。
该研究结果表明羊蹄根际土壤微生物多样性丰富，群落数量更多，羊蹄具有土壤改良作用。
Rumex patientia (R. patientia) is the second largest genus Polygonum genussorrel, there are about150species. R. patientia is rich in our country, with26species,distributed in provinces and regions. R. patientia contained in the "Shen Nong'sHerbal Classic". The herbal medicine in our country folk be used to stop bleeding andtreat scabies. Modern pharmacological studies have shown that it has anti-fungal,anti-tumor, anti-viral and anti-oxidative effects. Chemical analysis showed it has arich anthraquinone, tannic acid, naphthalene derivatives. In this thesis, Firstly, Wediscussed R. patientia rhizosphere and non-rhizosphere soils microbial diversity inJilin City four habitats using microbial culture and non-culture methods; secondly, wediscussed R. patientia rhizosphere and non-rhizosphere soils microorganisms inChangchun City two habitats using microbial culture method; Finally, we analyzedsoil microorganisms of R. patientia of similar habitats in Changchun and Jilin City.
The main results are listed as following:
1. This study suggested that the rhizosphere microorganisms mainly becomposed of bacteria, actinomycetes, followed by fungi least. The microbialpopulation is more in rhizosphere soils compared to non-rhizosphere soils of R.patientia of four habitats in Jilin City.
(1) The principal component analysis (PCA) biplot displayed that azotobacterand ammonifier was the strongest determinant for rhizosphere soils, and total bacteriaand actinomycetes were also the important factor for rhizosphere soils; Whereassulfur bacteria, total bacteria and ammonifier was the strongest determinant fornon-rhizosphere soils and actinomycetes and fungi were also the important factor fornon-rhizosphere soils.
(2) Pearson correlation coefficient analysis showed R. patientia rhizosphere ofJilin four habitats actinomycetes richness was significantly negatively correlated withmoisture content and humic matter, the abundance of ammonifier was significantlynegatively correlated with total P, azotobacter richness was very significant negativecorrelated with total P. In the non-rhizosphere soil, the total bacteria richness wassignificantly negatively correlated with total K, actinomycetes richness wassignificantly positively correlated with pH, and azotobacter richness weresignificantly negatively correlated with pH.
2. This study suggested that the microbial population is more in rhizospherecompared to non-rhizosphere soils of R. patientia of two habitats in Changchun City.
3. We compared rhizosphere soil microorganisms of R. patientia of similarhabitats between Jilin and Changchun City, These results show that the numbers ofrhizosphere soil microorganisms of R. patientia at habitats near water and sunnyhabitats in Jilin City were higher than Changchun City.
4. Our results indicate that unique clones in rhizosphere and non-rhizospheresoils are more than common clones in both of them in4different habitats in Jilin by16SrDNA technology, Furthermore, the unique clones in rhizosphere soil are morethan that in non-rhizosphere soil. The study results show that rhizosphere microbialdiversity is rich in R. patientia of rhizosphere soil and it has more number ofcommunities in rhizosphere soil, it has soil improvement effect.
本论文首先采用微生物培养及非培养的方法研究吉林市4个生境羊蹄根际与非根际土壤微生物多样性；其次采用微生物培养的方法研究长春市2个生境羊蹄根际与非根际土壤微生物；最后对吉林市与长春市相似生境的羊蹄根际土壤微生物进行了比较分析。
主要结果如下：
1.采用微生物培养的方法研究吉林市4个生境羊蹄根际与非根际土壤微生物主要由细菌、放线菌组成，真菌最少。根际与非根际土壤相比，微生物群落更多。
(1)主成分分析显示自生固氮菌、氨化细菌是根际土壤的最主要影响因子，另外，总细菌和放线菌也是重要的影响因子。硫化细菌，总细菌和氨化细菌是非根际土壤的最主要影响因子，另外，放线菌和真菌也是重要的影响因子。
(2)皮尔森相关系数分析显示吉林市4个生境羊蹄根际土壤放线菌的丰富度与湿度及腐殖质呈显著负相关，氨化细菌的丰富度与总P含量呈显著负相关，自生固氮菌的丰富度与总P含量呈非常显著负相关。在非根际土壤，总细菌的丰富度与总K含量呈显著负相关，放线菌的丰富度与pH呈显著正相关，自生固氮菌的丰富度与pH呈显著负相关。
2.长春市2个生境羊蹄根际土壤微生物群落数量多于非根际土壤。
3.吉林市与长春市两个相似生境的羊蹄根际土壤微生物比较分析结果表明，近水生境、向阳生境羊蹄根际土壤微生物数量吉林市均高于长春市。
4.采用16SrDNA技术研究吉林市4个不同生境下羊蹄根际与非根际土壤特有的克隆多于共有的克隆；并且根际土壤特有的克隆多于非根际土壤。
该研究结果表明羊蹄根际土壤微生物多样性丰富，群落数量更多，羊蹄具有土壤改良作用。
Rumex patientia (R. patientia) is the second largest genus Polygonum genussorrel, there are about150species. R. patientia is rich in our country, with26species,distributed in provinces and regions. R. patientia contained in the "Shen Nong'sHerbal Classic". The herbal medicine in our country folk be used to stop bleeding andtreat scabies. Modern pharmacological studies have shown that it has anti-fungal,anti-tumor, anti-viral and anti-oxidative effects. Chemical analysis showed it has arich anthraquinone, tannic acid, naphthalene derivatives. In this thesis, Firstly, Wediscussed R. patientia rhizosphere and non-rhizosphere soils microbial diversity inJilin City four habitats using microbial culture and non-culture methods; secondly, wediscussed R. patientia rhizosphere and non-rhizosphere soils microorganisms inChangchun City two habitats using microbial culture method; Finally, we analyzedsoil microorganisms of R. patientia of similar habitats in Changchun and Jilin City.
The main results are listed as following:
1. This study suggested that the rhizosphere microorganisms mainly becomposed of bacteria, actinomycetes, followed by fungi least. The microbialpopulation is more in rhizosphere soils compared to non-rhizosphere soils of R.patientia of four habitats in Jilin City.
(1) The principal component analysis (PCA) biplot displayed that azotobacterand ammonifier was the strongest determinant for rhizosphere soils, and total bacteriaand actinomycetes were also the important factor for rhizosphere soils; Whereassulfur bacteria, total bacteria and ammonifier was the strongest determinant fornon-rhizosphere soils and actinomycetes and fungi were also the important factor fornon-rhizosphere soils.
(2) Pearson correlation coefficient analysis showed R. patientia rhizosphere ofJilin four habitats actinomycetes richness was significantly negatively correlated withmoisture content and humic matter, the abundance of ammonifier was significantlynegatively correlated with total P, azotobacter richness was very significant negativecorrelated with total P. In the non-rhizosphere soil, the total bacteria richness wassignificantly negatively correlated with total K, actinomycetes richness wassignificantly positively correlated with pH, and azotobacter richness weresignificantly negatively correlated with pH.
2. This study suggested that the microbial population is more in rhizospherecompared to non-rhizosphere soils of R. patientia of two habitats in Changchun City.
3. We compared rhizosphere soil microorganisms of R. patientia of similarhabitats between Jilin and Changchun City, These results show that the numbers ofrhizosphere soil microorganisms of R. patientia at habitats near water and sunnyhabitats in Jilin City were higher than Changchun City.
4. Our results indicate that unique clones in rhizosphere and non-rhizospheresoils are more than common clones in both of them in4different habitats in Jilin by16SrDNA technology, Furthermore, the unique clones in rhizosphere soil are morethan that in non-rhizosphere soil. The study results show that rhizosphere microbialdiversity is rich in R. patientia of rhizosphere soil and it has more number ofcommunities in rhizosphere soil, it has soil improvement effect.
引文
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[1]郑水庆.酸模属药用植物的生药鉴定与资源利用研究[D].第二军医大学,2001.
[2]许光辉,郑洪元.土壤微生物分析方法手册[M].北京:农业出版社,1986.
[3] Jones, D. L., A. Hodge, and Y. Kuzyakov. Plant and mycorrhizal regulation ofrhizodeposition[J]. New Phytologist,2004,163(3):459-480.
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[5] Govaerts, B., M. Mezzalama, K. D. Sayre, J. Crossa, K. Lichter, V. Troch, K.Vanherck, P. De Corte, and J. Deckers. Long-term consequences of tillage，residue management， and crop rotation on selected soil micro-flora groups inthe subtropical highlands[J]. Applied Soil Ecology,2008,38(3):197-210.
[6] Karthikeyan, B., C. A. Jaleel, G. M. A. Lakshmanan, and M. Deiveekasundaram.Studies on rhizosphere microbial diversity of some commercially importantmedicinal plants[J]. Colloids and Surfaces B-Biointerfaces,2008,62(1):143-145.
[7]邵玉琴,赵吉,包青海.库布齐固定沙丘土壤微生物生物量的垂直分布研究[J].中国沙漠,2001,21(1):88-92.
[1] Becker, J. M., T. Parkin, C. H. Nakatsu, J. D. Wilbur, and A. Konopka. Bacterialactivity, community structure, and centimeter-scale spatial heterogeneity incontaminated soil[J]. Microbial Ecology,2006,51(2):220-231.
[2] Mummey, D. L. and M. C. Rillig. Spatial characterization of arbuscularmycorrhizal fungal molecular diversity at the submetre scale in a temperategrassland[J]. Fems Microbiology Ecology,2008,64(2):260-270.
[3] Sey, B. K., A. M. Manceur, J. K. Whalen, E. G. Gregorich, and P. Rochette.Small-scale heterogeneity in carbon dioxide, nitrous oxide and methaneproduction from aggregates of a cultivated sandy-loam soil[J]. Soil Biology andBiochemistry,2008,40(9):2468-2473.
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[5] Schloss PD, Westcott SL, Ryabin T, Hall JR, Hartmann M, HollisterEB,Lesniewski RA, Oakley BB, Parks DH, Robinson CJ, Sahl JW, StresB,Thallinger GG, Van Horn DJ, Weber CF Introducing mothur: Open-Source，Platform-Independent, Community-Supported Software for Describing andComparing Microbial Communities[J]. Applied and Environmental Microbiology,2009,75(23):7537-7541.
[6] Shi JY, Yuan XF, Lin HR, Yang YQ, Li ZY. Differences in Soil Properties andBacterial Communities between the Rhizosphere and Bulk Soil and amongDifferent Production Areas of the Medicinal Plant Fritillaria thunbergii[J]. Int JMol Sci,2011,12(6):3770-3785.
[7] Lin YT, Lin CP, Chaw SM, Whitman WB, Coleman DC, Chiu CY Bacterialcommunity of very wet and acidic subalpine forest and fire-induced grasslandsoils[J]. Plant and Soil,2010,332(1-2):417-427.
[8] Chan OC, Yang XD, Fu Y, Feng ZL, Sha LQ, Casper P, Zou XM.16S rRNA geneanalyses of bacterial community structures in the soils of evergreen broad-leavedforests in south-west China[J]. FEMS Microbiol Ecol,2006,58(2):247-259.
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[1]郑水庆.酸模属药用植物的生药鉴定与资源利用研究[D].第二军医大学,2001.
[2]许光辉,郑洪元.土壤微生物分析方法手册[M].北京:农业出版社,1986.
[3] Jones, D. L., A. Hodge, and Y. Kuzyakov. Plant and mycorrhizal regulation ofrhizodeposition[J]. New Phytologist,2004,163(3):459-480.
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[6] Karthikeyan, B., C. A. Jaleel, G. M. A. Lakshmanan, and M. Deiveekasundaram.Studies on rhizosphere microbial diversity of some commercially importantmedicinal plants[J]. Colloids and Surfaces B-Biointerfaces,2008,62(1):143-145.
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[1] Becker, J. M., T. Parkin, C. H. Nakatsu, J. D. Wilbur, and A. Konopka. Bacterialactivity, community structure, and centimeter-scale spatial heterogeneity incontaminated soil[J]. Microbial Ecology,2006,51(2):220-231.
[2] Mummey, D. L. and M. C. Rillig. Spatial characterization of arbuscularmycorrhizal fungal molecular diversity at the submetre scale in a temperategrassland[J]. Fems Microbiology Ecology,2008,64(2):260-270.
[3] Sey, B. K., A. M. Manceur, J. K. Whalen, E. G. Gregorich, and P. Rochette.Small-scale heterogeneity in carbon dioxide, nitrous oxide and methaneproduction from aggregates of a cultivated sandy-loam soil[J]. Soil Biology andBiochemistry,2008,40(9):2468-2473.
[1] Woese C R. Bacterial evolution[J].Microbiol Rev,1987,51:221-217.
[2] Hugenholtz P, Goebel B M,Pace N R, Minireview:impact of culture-independentstudies on the emerging phylogenetic view of bacterial diversity[J]. JBacteriol,1998,180(18):4765-4774.
[3] Rappe M S, Giovannoni S J. The uncultured microbial majority[J]. Annu RevMicrobiol,2003,57:369-394.
[4] Jia X, Han S-j, Zhao Y-h, Zhou Y-m. Comparisons of extraction and purificationmethods of soil microorganism DNA from rhizosphere soil[J]. Journal of ForestryResearch,2006:17(1):31-34.
[5] Schloss PD, Westcott SL, Ryabin T, Hall JR, Hartmann M, HollisterEB,Lesniewski RA, Oakley BB, Parks DH, Robinson CJ, Sahl JW, StresB,Thallinger GG, Van Horn DJ, Weber CF Introducing mothur: Open-Source，Platform-Independent, Community-Supported Software for Describing andComparing Microbial Communities[J]. Applied and Environmental Microbiology,2009,75(23):7537-7541.
[6] Shi JY, Yuan XF, Lin HR, Yang YQ, Li ZY. Differences in Soil Properties andBacterial Communities between the Rhizosphere and Bulk Soil and amongDifferent Production Areas of the Medicinal Plant Fritillaria thunbergii[J]. Int JMol Sci,2011,12(6):3770-3785.
[7] Lin YT, Lin CP, Chaw SM, Whitman WB, Coleman DC, Chiu CY Bacterialcommunity of very wet and acidic subalpine forest and fire-induced grasslandsoils[J]. Plant and Soil,2010,332(1-2):417-427.
[8] Chan OC, Yang XD, Fu Y, Feng ZL, Sha LQ, Casper P, Zou XM.16S rRNA geneanalyses of bacterial community structures in the soils of evergreen broad-leavedforests in south-west China[J]. FEMS Microbiol Ecol,2006,58(2):247-259.
[9] Fierer N, Jackson RB. The diversity and biogeograPhy of soil bacterialcommunities[J]. Proc Natl Acad Sci U S A,2006,103(3):626-631.
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