枯草芽孢杆菌菌剂对烟草根际土壤细菌群落的影响
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- 英文论文题名：Impact of biocontrol agent Bacillus subtilis on bacterial communities in tobacco rhizospheric soil
- 论文作者：计思贵 ; 焦永鸽 ; 张立猛 ; 代快 ; 李江舟 ; 崔永和 ; 赵进龙
- 英文论文作者：JIAO Yongge ; JI Sigui ; ZHANG Limeng ; LI Jiangzhou ; DAI kuai ; CUI yonghe ; ZHAO jinlong ; Yunnan Tobacco Company ; Yuxi Branch
- 年：2016
- 作者机构：云南省烟草公司玉溪市公司;
- 论文关键词：枯草芽孢杆菌 ; 群落结构 ; 16S rRNA基因 ; ARDRA
- 英文论文关键词：Bacillus subtilis ; community composition ; 16S rRNA genes ; ARDRA
- 会议召开时间：2016-12-01
- 会议录名称：中国烟草学会2016年度优秀论文汇编——烟草农业主题
- 语种：中文
- 分类号：S482.292;S154.3
- 学会代码：ZGYG
- 学会名称：中国烟草学会
- 页数：10
- 文件大小：812k
- 原文格式：D
摘要
通过构建16S r RNA克隆文库以及核糖体DNA扩增片段酶切分析(ARDRA)的方法,以同一种植烟草但不使用促生剂土壤为对照,研究施用生物防治剂枯草芽孢杆菌菌剂对烟草根际土壤细菌群落结构以及多样性的影响。采用文库库容值(C)、Shannon多样性指数(H’)、Pielou均匀度指数(E)和Margalef丰富度指数(R)对细菌多样性进行评价。系统发育表明,2个样品均检测到以下12个细菌类群:酸杆菌门(Acidobacteria)、变形菌门(α-、β-、δ-、γ-Proteobacteria)、浮霉菌门(Planctomycetes)、厚壁菌门(Firmicutes)、硝化螺旋菌门(Nitrospirae)、芽单胞菌门(Gemmatimonadetes)、放线菌门(Actinobacteria)、绿弯菌门(Chloroflexi)和拟杆菌门(Bacteroidetes);但各细菌类群的结构组成及所占比例在不同样品中有较大差别。对照土壤中优势菌群为Acidobacteria(27.11%)和Proteobacteria(26.51%);处理土壤中则为Proteobacteria(37.98%)和Acidobacteria(29.61%);菌剂处理后土壤中,Gammaproteobacteria和Alphaproteobacteria所占比例明显提高,而Betaproteobacteria、Planctomycetes和Firmicutes 3个菌群的数量则相对降低。多样性分析表明2个土壤样品均具有丰富的多样性,且经枯草芽孢杆菌菌剂处理后,土壤细菌多样性指数和丰富度指数均提高。
Using non-treated soil as control, the impact of inoculation with the biocontrol agent Bacillus subtilis on bacterial communities and bacterial diversity in rhizospheric soil of Nicotiana tabacum L. was assessed by constructing a 16 S r RNA gene clone library and conducting amplified ribosomal DNA restriction analysis(ARDRA). The bacterial diversity was evaluated by Coverage value(C), Shannon–Weiner index(H), Pielou evenness index(E) and Margalef richness index(R). Phylogenetic analysis revealed that the inoculation significantly affected the composition of bacterial communities in tobacco rhizospheric soil. Twelve bacterial groups including Acidobacteria, Proteobacteria(including α-, β-, δ-, γ-Proteobacteria), Planctomycetes, Firmicutes, Nitrospirae, Gemmatimonadetes, Actinobacteria and Bacteroidetes were detected to be shared by inoculated soil and control soil. The community composition and proportion of different bacteria in communities showed significant variation between the two samples. The dominant bacteria were Acidobacteria(27.11%) and Proteobacteria(26.51%) in non-treated soil, while in the inoculated soil Proteobacteria(37.98%) and Acidobacteria(29.61%) were dorminant. Bacillus subtilis inoculation increased the numbers of Gamma-proteobacteria and Alpha-proteobacteria but reduced the numbers of Beta-proteobacteria, Planctomycetes and Firmicutes. Diversity analysis showed that both soil samples had rich diversity, and soil bacterial Shannon–Weiner index and Margalef richness index were promoted after inoculation.
Using non-treated soil as control, the impact of inoculation with the biocontrol agent Bacillus subtilis on bacterial communities and bacterial diversity in rhizospheric soil of Nicotiana tabacum L. was assessed by constructing a 16 S r RNA gene clone library and conducting amplified ribosomal DNA restriction analysis(ARDRA). The bacterial diversity was evaluated by Coverage value(C), Shannon–Weiner index(H), Pielou evenness index(E) and Margalef richness index(R). Phylogenetic analysis revealed that the inoculation significantly affected the composition of bacterial communities in tobacco rhizospheric soil. Twelve bacterial groups including Acidobacteria, Proteobacteria(including α-, β-, δ-, γ-Proteobacteria), Planctomycetes, Firmicutes, Nitrospirae, Gemmatimonadetes, Actinobacteria and Bacteroidetes were detected to be shared by inoculated soil and control soil. The community composition and proportion of different bacteria in communities showed significant variation between the two samples. The dominant bacteria were Acidobacteria(27.11%) and Proteobacteria(26.51%) in non-treated soil, while in the inoculated soil Proteobacteria(37.98%) and Acidobacteria(29.61%) were dorminant. Bacillus subtilis inoculation increased the numbers of Gamma-proteobacteria and Alpha-proteobacteria but reduced the numbers of Beta-proteobacteria, Planctomycetes and Firmicutes. Diversity analysis showed that both soil samples had rich diversity, and soil bacterial Shannon–Weiner index and Margalef richness index were promoted after inoculation.
引文
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[6]Zhao Z-X(赵志祥),Luo K(罗坤),Chen G-H(陈国华),et al.Analysis of bacterial diversity in rhizosphere of cucumber in greenhouse by the methods of metagenomic end-random sequencing and 16S r RNA technology.Acta Ecologica Sinica(生态学报),2010,30(14):3849-3857(in Chinese)
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[19]Liu W-Q(刘玮琦),Mao Z-C(茆振川),Yang Y-H(杨宇红),et al.Analysis of soil bacterial diversity by using the 16S r RNA gene Library.Acta Microbiologica Sinica(微生物学报),2008,48(10):1344-1350(in Chinese)
[20]Chen H,Xiao X,Jun W,et al.Antagonistic effects of volatiles generated by Bacillus subtilis on spore germination and hyphal growth of the plant pathogen,Botrytis cinerea.Biotechnology Letters,2008,30:919-923
[21]Zhou C(周翠),Qiao L-Q(乔鲁芹),Jin J(金静),et al.Preliminary study on the inhibitory effect of the volatile substances produced by Bacillus subtilis.Chinese Journal of Pesticide Science(农药学学报),2011,13(2):201-204(in Chinese)
[22]Zhang C-S(张成省),Kong F-Y(孔凡玉),Liu Z-K(刘朝科),et al.Tobacco growth enhancement and disease protection by the volatile-producing Bacillus subtilis Tpb55.Chinese Journal of Biological Control(中国生物防治),2009,25(3):245-249(in Chinese)
[23]Liu F-C(刘方春),Xing S-J(邢尚军),Ma H-L(马海林),et al.Effects of inoculating plant growth-promoting rhizobacteria on the biological characteristics of walnut(Juglans regia)rhizosphere soil under drought condition[EB/OL].(2014-03-12)[2014-4-22].http://www.cnki.net/kcms/doi/10.13287/j.1001-9332.20140305.0014.html
[24]Schwieger F,Tebbe CC.Effect of field inoculation with Sinorhizobium meliloti L33 on the composition of bacterial communities in rhizospheres of a target plant(Medicago sativa)and a non-target plant(Chenopodium album)—linking of 16S r RNA gene-based single-strand conformation polymorphism community profiles to the diversity of cultivated bacteria.Applied and Environmental Microbiology,2000,66(8):3556-3565
[25]Trabelsi D,Mengoni A,Ammar HB,et al.Efect of on-field inoculation of Phaseolus vulgaris with rhizobia on soil bacterial communities.FEMS Microbiology Ecology,2011,77(1):211-222
[26]Zhang BG,Bai ZH,Hoefel D,et al.Assessing the impact of the biological control agent Bacillus thuringiensis on the indigenous microbial community within the pepper plant phyllosphere.FEMS Microbiology Letters,2008,284(1):102-108
[27]Antoun H,Beauchamp CJ,Goussard N,et al.Potential of Rhizobium and Bradyrhizobium species as plant growth promoting rhizobacteria on non-legumes:Effect on radishes(Raphanus sativus L.).Plant and Soil,1998,204:57-67
[28]Zhong Z-T(钟增涛),Shen Q-R(沈其荣),Sun X-H(孙晓红),et al.Role of Rhizobium in wheat-astragalus mixed cropping system.Chinese Journal of Applied Ecology(应用生态学报),2003,14(2):187-190(in Chinese)
[29]Palaniappan P,Chauhan PS,Saravanan VS,et al.Isolation and characterization of plant growth promoting endophytic bacterial isolates from root nodule of Lespedeza sp.Biology and Fertility of Soils,2010,46(8):807-816
[30]Clercq DD,Trappen SV,Cleenwerck I,et al.Rhodanobacter spathiphylli sp.nov.,a gammaproteobacterium isolated from the roots of Spathiphyllum plants grown in a compost-amended potting mix.International Journal of Systematic and Evolutionary Microbiology,2006,56(8):1755-1759
[2]Chen X-R(陈秀蓉),Nan Z-B(南志标).Bacterial diversity and its role in agricultural ecosystems.Pratacultural Science(草业科学),2002,19(9):34-38(in Chinese)
[3]Zhang Y-W(张云伟),Xu Z(徐智),Tang L(汤利),et al.Effects of different organic fertilizers on the microbes in rhizospheric soil of flue-cured tobacco.Chinese Journal of Applied Ecology(应用生态学报),2013,24(9):2551-2556(in Chinese)
[4]Lei J-L(雷娟利),Zhou Y-H(周艳虹),Ding J(丁桔),et al.Effect of continuous cropping of different vegetables on DNA polymorphorism of soil bacterial.Scientia Agricultura Sinica(中国农业科学),2005,38(10):2076-2083(in Chinese)
[5]Sang MK,Kim DK.Plant growth-promoting rhizobacteria suppressive to Phytophthora blight affect microbial activities and communities in the rhizosphere of pepper(Capsicum annuum L.)in the field.Applied Soil Ecology.2012,62:88-97
[6]Zhao Z-X(赵志祥),Luo K(罗坤),Chen G-H(陈国华),et al.Analysis of bacterial diversity in rhizosphere of cucumber in greenhouse by the methods of metagenomic end-random sequencing and 16S r RNA technology.Acta Ecologica Sinica(生态学报),2010,30(14):3849-3857(in Chinese)
[7]Xu Y-G(徐永刚),Yu W-T(宇万太),Ma Q(马强),et al.Effects of long-term fertilizations on microbial biomass C and N and bacterial community structure in an aquic brown soil.Chinese Journal of Applied Ecology(应用生态学报),2010,21(8):2078-2085(in Chinese)
[8]Trabelsi D,Mhamdi R.Microbial inoculants and their impact on soil microbial communities:a review.Bio Med Research International.2013,2013:863240
[9]Dawar S,Wahab S,Tariq M,et al.Application of Bacillus species in the control of root rot diseases of crop plants.Archives of Phytopathology and Plant Protection.2010,43(4):412-418
[10]Huang Z-X(黄祖新),Huang Z(黄镇),Ye B-Y(叶冰莹),et al.Comparison of culture-dependent and-independent approaches for diversity analysis of soil bacteria in the rhizosphere of sugarcane.Chinese Journal of Applied&Environmental Biology(应用与环境生物学报),2011,17(5):742-746(in Chinese)
[11]Amans RI,Luding E,Schleifer KH.Phylogenetic identification and in situ detection of individual microbial cells without cultivation.Microbiology and Molecular Biology Reviews,1995,59(1):143-169
[12]Weidner S,Arnold W,Puhler A.Diversity of uncultured microorganisms associated with the seagrass Halophila stipulacea estimated by restriction fragment length polymorphism analysis of PCR-amplified 16S r RNA genes.Applied and Environmental Microbiology,1996,62(3):766-771
[13]Zhang C-S(张成省),Kong F-Y(孔凡玉),Li D-C(李多川),et al.Screening of antagonistic bacteria from phylloshere and its resistance to Alternaria alternata.Acta Tabacaria Sinica(中国烟草学报),2005,11(4):17-20(in Chinese)
[14]Zhang C-S(张成省),Kong F-Y(孔凡玉),Guan X-H(关小红),et al.Identification and antagonistic activity of tobacco phyllosphere bacteria strain Tpb55.Chinese Journal of Biological Control(中国生物防治),2008,24(1):63-68
[15]Schloss PD,Handelsman J.Status of the microbial census.Microbiology and Molecular Biology Reviews,2004,68(4):686-691
[16]Zhang J-P(张建萍),Dong N-Y(董乃源),Yu H-B(余浩滨),et al.Bacteria diversity in paddy field soil by16S r RNA-RFLP analysis in Ningxia.Biodiversity Science(生物多样性),2008,16(6):586-592(in Chinese)
[17]Jiang DM,Sun YH,Li D,et al.Characterization of bacteria community associated with soil arsenic and sulphate contamination based on 16S r RNA gene sequences.Microbiology China,2011,38(10):1592-1601
[18]Bai L(白蓝),Zhao M-W(赵明文),Jia J-W(贾军伟),et al.Analysis of soil bacterial community composition by 16S r RNA clone library sampling from transgenic carnation.Microbiology China(微生物学通报),2012,39(4):435-447(in Chinese)
[19]Liu W-Q(刘玮琦),Mao Z-C(茆振川),Yang Y-H(杨宇红),et al.Analysis of soil bacterial diversity by using the 16S r RNA gene Library.Acta Microbiologica Sinica(微生物学报),2008,48(10):1344-1350(in Chinese)
[20]Chen H,Xiao X,Jun W,et al.Antagonistic effects of volatiles generated by Bacillus subtilis on spore germination and hyphal growth of the plant pathogen,Botrytis cinerea.Biotechnology Letters,2008,30:919-923
[21]Zhou C(周翠),Qiao L-Q(乔鲁芹),Jin J(金静),et al.Preliminary study on the inhibitory effect of the volatile substances produced by Bacillus subtilis.Chinese Journal of Pesticide Science(农药学学报),2011,13(2):201-204(in Chinese)
[22]Zhang C-S(张成省),Kong F-Y(孔凡玉),Liu Z-K(刘朝科),et al.Tobacco growth enhancement and disease protection by the volatile-producing Bacillus subtilis Tpb55.Chinese Journal of Biological Control(中国生物防治),2009,25(3):245-249(in Chinese)
[23]Liu F-C(刘方春),Xing S-J(邢尚军),Ma H-L(马海林),et al.Effects of inoculating plant growth-promoting rhizobacteria on the biological characteristics of walnut(Juglans regia)rhizosphere soil under drought condition[EB/OL].(2014-03-12)[2014-4-22].http://www.cnki.net/kcms/doi/10.13287/j.1001-9332.20140305.0014.html
[24]Schwieger F,Tebbe CC.Effect of field inoculation with Sinorhizobium meliloti L33 on the composition of bacterial communities in rhizospheres of a target plant(Medicago sativa)and a non-target plant(Chenopodium album)—linking of 16S r RNA gene-based single-strand conformation polymorphism community profiles to the diversity of cultivated bacteria.Applied and Environmental Microbiology,2000,66(8):3556-3565
[25]Trabelsi D,Mengoni A,Ammar HB,et al.Efect of on-field inoculation of Phaseolus vulgaris with rhizobia on soil bacterial communities.FEMS Microbiology Ecology,2011,77(1):211-222
[26]Zhang BG,Bai ZH,Hoefel D,et al.Assessing the impact of the biological control agent Bacillus thuringiensis on the indigenous microbial community within the pepper plant phyllosphere.FEMS Microbiology Letters,2008,284(1):102-108
[27]Antoun H,Beauchamp CJ,Goussard N,et al.Potential of Rhizobium and Bradyrhizobium species as plant growth promoting rhizobacteria on non-legumes:Effect on radishes(Raphanus sativus L.).Plant and Soil,1998,204:57-67
[28]Zhong Z-T(钟增涛),Shen Q-R(沈其荣),Sun X-H(孙晓红),et al.Role of Rhizobium in wheat-astragalus mixed cropping system.Chinese Journal of Applied Ecology(应用生态学报),2003,14(2):187-190(in Chinese)
[29]Palaniappan P,Chauhan PS,Saravanan VS,et al.Isolation and characterization of plant growth promoting endophytic bacterial isolates from root nodule of Lespedeza sp.Biology and Fertility of Soils,2010,46(8):807-816
[30]Clercq DD,Trappen SV,Cleenwerck I,et al.Rhodanobacter spathiphylli sp.nov.,a gammaproteobacterium isolated from the roots of Spathiphyllum plants grown in a compost-amended potting mix.International Journal of Systematic and Evolutionary Microbiology,2006,56(8):1755-1759
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