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作物学报 ›› 2015, Vol. 41 ›› Issue (02): 308-317.doi: 10.3724/SP.J.1006.2015.00308

• 耕作栽培·生理生化 • 上一篇    下一篇

不同连作年限野生地黄根际土壤微生物群落多样性分析

吴林坤1,2,黄伟民1,2,**,王娟英1,2,**,吴红淼1,2,陈军1,2,秦贤金1,2,张重义2,林文雄1,2,*   

  1. 1 福建农林大学生命科学学院, 福建福州 350002; 2 福建农林大学农业生态研究所, 福建福州 350002
  • 收稿日期:2014-04-18 修回日期:2014-12-19 出版日期:2015-02-12 网络出版日期:2014-12-29
  • 通讯作者: 林文雄, E-mail: wenxiong181@163.com
  • 基金资助:

    本研究由国家重点基础研究发展计划(973计划)项目(2012CB126309), 国家自然基金联合基金项目(U1205021)和国家自然科学基金项目(81303170)资助。

Diversity Analysis of Rhizosphere Microflora of Wild R. glutinosa Grown in Monocropping for Different Years

WU Lin-Kun1,2,HUANG Wei-Min1,2,**,WANG Juan-Ying1,2,**,WU Hong-Miao1,2,CHEN Jun1,2,QIN Xian-Jin1,2,ZHANG Zhong-Yi2,LIN Wen-Xiong1,2,*   

  1. 1 College of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China; 2 Agricultural Agroecological Institute, Fujian Agriculture and Forestry University, Fuzhou 350002, China
  • Received:2014-04-18 Revised:2014-12-19 Published:2015-02-12 Published online:2014-12-29
  • Contact: 林文雄, E-mail: wenxiong181@163.com

摘要:

以野生地黄为试验材料,设置野生地黄头茬土壤、重茬土壤、原茬土壤等处理,未种植任何作物为对照,于块根膨大中期采集土样,通过磷脂脂肪酸法(PLFA)和末端限制性片段长度多态性(T-RFLP)技术,分析不同连作年限野生地黄的根际微生物生物量和群落结构变化。PLFA分析结果表明,不同处理情况下地黄根际土壤微生物群落结构存在明显差异,与头茬地黄根际土壤相比,重茬地黄土壤微生物总量显著下降,并且细菌/真菌比例下降。T-RFLP分析结果表明不同连作年限地黄根际土壤细菌群落结构存在一定差异,野生状态地黄土壤和头茬土壤菌群较为相似,变形菌门和厚壁菌门占据优势地位。野生状态地黄和头茬地黄根际富含Bacillus、Pseudomonas等有益生防菌,而重茬地黄根际土壤滋生大量病原菌如Clostridium sp.、Flexibacter polymorphus、Clostridium ghoni,有益菌群和纤维素降解菌群减少,qRT-PCR定量分析也显示野生状态地黄和头茬地黄土壤中假单胞菌数量都显著高于重茬地黄土壤。总之,野生地黄存在连作障碍问题,导致野生地黄根际有益菌数量减少而病原菌大量滋生,从而降低了野生地黄抵御病害的能力,使重茬野生地黄生长发育差,产量大幅降低。

关键词: 地黄, 磷脂脂肪酸, 末端限制性片段长度多态性, 微生物多样性, 植物根际

Abstract:

The soils sampled from the four different plots, including the newly planted, the two-year monocultured, the wild R. glutinosa and the control without growing R. glutinosa, were used to study the changes in microbial biomass and community composition using phospholipid fatty acid (PLFA) and terminal restriction fragment length polymorphism (T-RFLP) analyses. PLFA analysis indicated that the soil microbial community composition was significantly different among the R. glutinosa with different years of monoculture. Compared with the newly planted soil, the total PLFA content and the ratio of bacteria/fungus in two-year monocultured soil greatly declined. Further analysis by T-RFLP also displayed the distinct differences in rhizospheric bacterial community structure of R. glutinosa. The microbial compositions from the wild and the newly planted R. glutinosa soils tended to be more similar. It was found that the bacteria including Proteobacteria and Firmicutes were predominant in the wild and newly planted R. glutinosa soils. Some beneficial biocontrol bacteria (such as Bacillus, Pseudomonas, etc.) gathered in the rhizosphere of the wild and newly planted R. glutinosa. However, a large number of pathogenic bacteria bred in the rhizosphere of the two-year monocultured R. glutinosa, such as Clostridium sp., Flexibacter polymorphus and Clostridium ghoni, and the number of beneficial bacteria and cellulose degradation bacteria decreased. Furthermore, qRT-PCR analysis verified that the total number of Pseudomonas was much higher in the wild and newly planted R. glutinosa soils than in the two-year monocultured soil. In conclusion, the pathogenic microbes breed seriously in the rhizospheric soil of wild R. glutinosa under the monoculture regime, and yet the number of beneficial bacteria decline, resulting in weakened ability of wild R. glutinosa to resist the diseases so that the two-year monocultured wild R. glutinosa grows abnormally and its yield is decreased drastically.

Key words: Rehmannia glutinosa, PLFA, T-RFLP, Microbial diversity, Plant rhizosphere

[1]张重义, 林文雄. 药用植物的化感自毒作用与连作障碍. 中国生态农业学报, 2009, 17: 189–196

Zhang Z Y, Lin W X. Continuous cropping obstacle and allelopathic autotoxicity of medicinal plants. Chin J Eco-Agric, 2009, 17: 189–196 (in Chinese with English abstract)

[2]Butler J L, Williams M A, Bottomley P J, Myrold D D. Microbial community dynamics associated with rhizosphere carbon flow. Appl Environ Microbiol, 2003, 69: 6793–6800

[3]Doornbos R F, van Loon L C, Bakker P A H M. Impact of root exudates and plant defense signaling on bacterial communities in the rhizosphere: A review. Agron Sustain Dev, 2012, 32: 227–243

[4]Eisenhauer N, Scheu S, Jousset A. Bacterial diversity stabilizes community productivity. PLoS ONE, 2012, 7: e34517

[5]Mendes R, Kruijt M, de Bruijn I, Dekkers E, van der Voort M, Schneider J H, Piceno Y M, DeSantis T Z, Andersen G L, Bakker P A, Raaijmakers J M. Deciphering the rhizosphere microbiome for disease-suppressive bacteria. Science, 2011, 332: 1097-1100

[6]王明道, 吴宗伟, 原增艳, 陈红歌, 吴坤, 贾新成. 怀地黄连作对土壤微生物区系的影响. 河南农业大学学报, 2008, 42: 532–538

Wang M D, Wu Z W, Yuan Z Y, Chen H G, Wu K, Jia X C. Effects of Rehmannia glutinosa Libosch. continuous cropping on microbial communities. J Henan Agric Univ, 2008, 42: 532–538 (in Chinese with English abstract)

[7]van Bruggen A H, Semenov A M, Zelenev V V. Wavelike distributions of microbial populations along an artificial root moving through soil. Microb Ecol, 2000, 40: 250–259

[8]Amann R I, Ludwig W, Schleifer K H. Phylogenetic identification and in situ detection of individual microbial cells without cultivation. Microbiol Rev, 1995, 59: 143–169

[9]任南琪, 赵阳国, 高崇洋, 王爱杰. TRFLP在微生物群落结构与动态分析中的应用. 哈尔滨工业大学学报, 2007, 39: 552–556

Ren N Q, Zhao Y G, Gao C Y, Wang A J. Terminal restriction fragment length polymorphism: a powerful technique for characterizing microbial community structure and dynamics. J Harbin Inst Technol, 2007, 39: 552–556 (in Chinese with English abstract)

[10]Dunbar J, Ticknor L O, Kuske C R. Assessment of microbial diversity in four southwestern United States soils by 16S rRNA gene terminal restriction fragment analysis. Appl Environ Microbio1, 2000, 66: 2943–2950

[11]Kitts C L. Terminal restriction fragment patterns: a tool for comparing microbial communities and assessing community dynamics. Curr Issues Intest Microbiol, 2001, 2: 17–25

[12]鲍士旦. 土壤农化分析. 北京: 中国农业出版社, 2008

Bao S D. Soil and Agricultural Chemistry Analysis. Beijing: China Agriculture Press, 2008

[13]Green C T, Scow K M. Analysis of phospholipid fatty acids (PLFA) to characterize microbial communities in aquifers. Hydrogeol J, 2000, 8: 126–141

[14]刘波, 胡桂萍, 郑雪芳, 张建福, 谢华安. 利用磷脂脂肪酸(PLFAs)生物标记法分析水稻根际土壤微生物多样性. 中国水稻科学, 2010, 24: 278–288

LIU B, Hu G P, Zheng X F, Zhang J F, Xie H A. Analysis on microbial diversity in the rhizosphere of rice by phospholipid fatty acids biomarkers. Chin J Rice Sci, 2010, 24: 278–288 (in Chinese with English abstract)

[15]B??th E, Anderson T H. Comparison of soil fungal/bacterial ratios in a pH gradient using physiological and PLFA-based techniques. Soil Biol Biochem, 2003, 35: 955–963

[16]Johansen A, Olsson S. Using phospholipid fatty acid technique to study short-term effects of the biological control agent Pseudomonas fluorescens DR54 on the microbial microbiota in barley rhizosphere. Microb Ecol, 2005, 49: 272–281

[17]齐鸿雁, 薛凯, 张洪勋. 磷脂脂肪酸谱图分析方法及其在微生物生态学领域的应用. 生态学报, 2003, 23: 1577–1579

Qi H Y, Xue K, Zhang H X. Phospholipid fatty acid analysis and its applications in microbial ecology. Acta Ecol Sin, 2003, 23: 1577–1579 (in Chinese with English abstract)

[18]Joergensen R G, Potthoff M. Microbial reaction in activity, biomass and community structure after long-term continuous mixing of a grassland soil. Soil Biol Biochem, 2005, 37: 1249–1258

[19]张秋芳, 刘波, 林营志, 史怀, 杨述省, 周先治. 土壤微生物群落磷脂脂肪酸PLFA生物标记多样性. 生态学报, 2009, 29: 4127–4137

Zhang Q F, Liu B, Lin Y Z, Shi H, Yang S S, Zhou X Z. The diversity of phospholipid fatty acid (PLFA) biomarker for the microbial community in soil. Acta Ecol Sin, 2009, 29: 4127–4137 (in Chinese with English abstract)

[20]Zhou J, Bruns M A, Tiedje J M. DNA recovery from soils of diverse composition. Appl Environ Microbiol, 1996, 62: 316–322

[21]Tipayno S, Kim C G, Sa T. T-RFLP analysis of structural changes in soil bacterial communities in response to metal and metalloid contamination and initial phytoremediation. Appl Soil Ecol, 2012, 61: 137–146

[22]Wang M, Ahrné S, Antonsson M, Molin G. T-RFLP combined with principal component analysis and 16S rRNA gene sequencing: an effective strategy for comparison of fecal microbiota in infants of different ages. J Microbiol Meth, 2004, 59: 53–69

[23]Tan Y, Ji G. Bacterial community structure and dominant bacteria in activated sludge from a 70 degrees C ultrasound enhanced anaerobic reactor for treating carbazole-containing wastewater. Bioresource Technol, 2010, 101: 174–180

[24]彭有才, 刘挺, 赵俊杰, 孙曙光, 高峻, 吴福如, 刘国顺, 叶协锋. 连作对土壤性状影响的研究进展. 江西农业学报, 2009, 21: 100–103

Peng Y C, Liu T, Zhao J J, Sun S G, Gao J, Wu F R, Liu G S, Ye X F. Research advances in effect of continuous cropping on soil characteristics. Acta Agric Jiangxi, 2009, 21: 100–103 (in Chinese with English abstract)

[25]王茂胜, 姜超英, 潘文杰, 薛小平, 陈懿, 梁永江. 不同连作年限的植烟土壤理化性质与微生物群落动态研究. 安徽农业科学, 2008, 36: 5033–5034

Wang M S, Jiang C Y, Pan W J, Xue X P, Chen Y, Liang Y J. Studying on physico-chemical properties and microbiological community in tobacco growing soils under different continuous cropping years. J Anhui Agric Sci, 2008, 36: 5033–5034 (in Chinese with English abstract)

[26]郭红伟, 郭世荣, 刘来, 孙锦, 黄保健. 辣椒连作对土壤理化性状、植株生理抗性及离子吸收的影响. 土壤, 2012, 44: 1041–1047

Guo H W, Guo S R, Liu L, Sun J, Huang B J. Effects of continuous cropping on physical and chemical properties of soil, physiological resistance and ion absorption of pepper. Soil, 2012, 44: 1041–1047 (in Chinese with English abstract)

[27]Zhou X, Wu F. P-Coumaric acid influenced cucumber rhizosphere soil microbial communities and the growth of Fusarium oxysporum f. sp. cucumerinum Owen. PLoS ONE, 2012, 7: e48288

[28]Bhattacharyya P N, Jha D K. Plant growth-promoting rhizobacteria (PGPR): emergence in agriculture. World J Microbiol Biotechnol, 2012, 28: 1327–1350

[29]Viswanathan R, Samiyappan R. Induced systemic resistance by fluorescent pseudomonads against red rot disease of sugarcane caused by Colletotrichum falcatum. Crop Prot, 2002, 21: 1–10

[30]Haas D, Défago G. Biological control of soil-borne pathogens by fluorescent pseudomonads. Nat Rev Microbiol, 2005, 3: 307–319

[31]Jetiyanon K, Kloepper J W. Mixtures of plant growth-promoting rhizobacteria for induction of systemic resistance against multiple plant diseases. Biol Control, 2002, 24: 285–291

[32]Zhu Y, Li H, Zhou H, Chen G, Liu W. Cellulose and cellodextrin utilization by the celulolytic bacterium Cytophaga hutchisonii. Bioresource Technol, 2010, 101: 6432–6437

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