花生青枯病生防细菌的筛选与鉴定
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摘要
【目的】筛选花生青枯病的生物防治菌株。【方法】利用平板对峙法筛选对花生青枯病有拮抗作用的生物防治细菌菌株,利用盆栽试验筛选防治效果好的细菌菌株。【结果】从根际土壤中分离的300余株细菌中筛选获得7株对花生青枯病有拮抗作用的菌株。其中菌株Ab、A27和A38的抑菌直径超过10 mm,且其分泌物对该菌有抑制效果。将这3个菌株进一步进行盆栽试验,结果表明A38的防治效果最好,达到52.72%。通过生理生化及分子生物学鉴定,将A38菌株鉴定为绿脓假单胞杆菌(Pseudomonas aeruginosa)。【结论】筛选到一种花生青枯病菌拮抗效果较好的生防细菌,鉴定为绿脓假单胞杆菌。
[Objective]To Screen the biocontrol bacterial strains against bacterial wilt of peanuts. [Method]The biocontrol bacterial strains which have antagonistic effects on bacterial wilt of peanuts were screened by plate confrontation method and the bacterial strains with good control effect were screened by pot experiment.[Result]Seven strains with antagonistic effects on bacterial wilt of peanuts were screened from over 300 bacteria strains isolated from rhizosphere soil. Among them, strains Ab, A27 and A38 had a bacterial inhibition diameter of more than 10 mm, and their secretions had an inhibitory effect on the bacteria. The three strains were further subjected to pot experiment, and the results showed that A38 got the best control effect, reaching 52.72%. The strain A38 was identified as Pseudomonas aeruginosa by physiological, biochemical and molecular biological identification.[Conclusion]A biocontrol bacterium with good antagonistic effect against bacterial wilt of peanuts was screened and identified as Pseudomonas aeruginosa.
[Objective]To Screen the biocontrol bacterial strains against bacterial wilt of peanuts. [Method]The biocontrol bacterial strains which have antagonistic effects on bacterial wilt of peanuts were screened by plate confrontation method and the bacterial strains with good control effect were screened by pot experiment.[Result]Seven strains with antagonistic effects on bacterial wilt of peanuts were screened from over 300 bacteria strains isolated from rhizosphere soil. Among them, strains Ab, A27 and A38 had a bacterial inhibition diameter of more than 10 mm, and their secretions had an inhibitory effect on the bacteria. The three strains were further subjected to pot experiment, and the results showed that A38 got the best control effect, reaching 52.72%. The strain A38 was identified as Pseudomonas aeruginosa by physiological, biochemical and molecular biological identification.[Conclusion]A biocontrol bacterium with good antagonistic effect against bacterial wilt of peanuts was screened and identified as Pseudomonas aeruginosa.
引文
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[7]郭坚华,王玉菊,李瑾,任欣正.抑菌圈-定殖力双重测定法筛选青枯病生防细菌[J].植物病理学报,1996,26(1):49-54.DOI:10.13926/j.cnki.apps.1996.01.014.GUO J H,WANG Y J,LI J,REN X Z.Screen of biocontrol bacteria of plant wilt by inhibiting zones and root-colonizing capacity[J].Acta Phytophylacica Sinica,1996,26(1):49-54.
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[9]蒲小明,林壁润,郑奕雄,沈会芳.一株抗花生青枯病菌海洋放线菌的分类鉴定及其活性产物研究[J].广东农业科学,2011,38(16):62-65.doi:10.16768/j.issn.1004-874x.2011.16.039.PU X M,LIN B R,ZHENG Y X,SHEN H F.Studies on taxonomic identification of a marine actinomycete against Pseudomonas solanacearum and its active product[J].Guangdong Agricultural Sciences,2011,38(16):62-65.doi:10.16768/j.issn.10 04-874x.2011.16.039.
[10]樊胜南,喻国辉,陈燕红,黎永坚,陈川雁.枯草芽孢杆菌TR21可湿性粉剂对香蕉根际土壤微生物的影响[J].广东农业科学,2018,45(10):68-73.doi:10.16768/j.issn.1004-874X.2018.10.011.FAN S N,YU G H,CHEN Y H,LI Y J,CHEN C Y.Influence of Bacillus subtilis strain TR21 wettable powder on the banana rhizosphere microorganisms[J].Guangdong Agricultural Sciences,2018,45(10):68-73.doi:10.16768/j.issn.1004-874X.2018.10.011.
[11]游春平,刘凤权.植物病原细胞研究技术[M].北京:中国农业出版社,2011.YOU C P,LIU F Q.Plant Pathogenic Cell Research Technology[M].Beijing:China Agricultural Press,2011.
[12]张建华,任欣正,方中达.青枯假单胞杆菌产生细菌素的研究[J].南京农业大学学报,1987,10(3):58-63.ZHANG J H,REN X Z,FANG Z D.Studies on bacteriocin production of Pseudomonas solanacearum[J].Journal of Nanjing agricultural university,1987,10(3):58-63.
[13]KOUMOUTSI A,CHEN X H,HENNE A,LIESEGANG H,HITZEROTH G,FRANKE P.VATER J.BORRISS R.Structural and functional characterization of gene-clusters directing nonribosomal synthesis of bioactive cycliclipopeptides in Bacillus amyloliquefaciens strains FZB42[J].Journal of Bacteriology,2004,186(4):1084-1096.doi:10.1128/JB.186.4.1084-1096.2004.
[14]刘波.青枯雷尔氏菌的生物学及其生物防治[M].福州:福建省农科院农业生物资源研究所农业微生物研究中心,2003.LIU B.Biology of Ralstonia solanacearum and its biological control[M].Fuzhou:Agricultural Microbiology Research Center,Institute of Agricultural Biological Resources,Fujian Academy of Agricultural Sciences,2003.
[15]喻国辉,程萍,王燕鹂,陈远凤,陈燕红,黎永坚.枯草芽孢杆菌TR21田间防治巴西蕉枯萎病的效果[J].中国生物防治学报,2010,26(4):497-500.doi:10.16409/j.cnki.2095-039x.2010.04.020.YU G H,CHENG P,WANG Y L,CHEN Y F,CHEN Y H,LI Y J.Control effect of bacillus subtilis Strain TR21 on panama disease of banana brazil(Musa spp.)in fields[J].Chinese Journal of Biological Control,2010,26(4):497-500.doi:10.16409/j.cnki.2095-039x.2010.04.020.
[16]王成义,闫茂仓,陈少波,管敏鑫,单乐州,艾为明,谢起浪,蔡延马奔.基于16S rRNA和recA香鱼鳗利斯顿氏菌的分离鉴定[J].海洋通报,2010,29(1):84-90.WANG C Y,YAN M C,CHEN S B,GUAN M X,SHAN L Z,AIW M,XIE Q L,CAI Y B.Isolation and identification of Listonella anguillarum from Plecoglossus altivelis based on 16S rRNA and recA[J].Marine science bulletin,2010,29(1):84-90.
[2]FRENCH E,KIM B S,RIVERA-Zuluaga K,IYER-Pascuzzi A S.Whole root transcriptomic analysis suggests a role for auxin pathways in resistance to Ralstonia solanacearum in tomato[J].Molecular PlantMicrobe Interactions:MPMI,2018,31(4):432-444.doi:10.1094/MPMI-08-17-0209-R.
[3]华丽霞,何炼,蒋秋平,曾华兰,叶鹏盛,韦树谷,黄玲,代顺冬.不同产区花生青枯病菌遗传多样性的分析[J].基因组学与应用生物学,2018,37(3):1349-1355.doi:10.13417/j.gab.037.001349.HUA L,HE L X,JIANG Q X,ZENG H G,YE P S G,WEI S L,HUANGL,DAI S D.Genetics diversity analysis of Ralstonia solanacearum on peanut from different producing areas[J].Genomics and Applied Biology,2018,37(3):1349-1355.
[4]周桂元,梁炫强,李一聪,李少雄.花生青枯病抗性鉴定及抗源分析,花生学报,2003,32(3):25-28.doi:10.14001/j.issn.1002-4093.2003.03.005.ZHOU GY,LIANG X Q,LI Y C,LI S X.Evaluation of peanut cultivars for resistance to bacterial wilt and analysis of family tree[J].Journal of Peanut Science,2003,32(3):25-28.doi:10.14001/j.issn.1002-4093.2003.03.005.
[5]罗宽,何昆,匡传富,周志成.三株拮抗细菌对烟青枯病的抑制效果[J].2002,18(4):185-187.DOI:10.16409/j.cnki.2095-039x.2002.04.010.LUO K,HE K,KUANG C F,ZHOU Z C.Studies on the inhibition of antagonistic bacteria to tobacco bacterial wilt,Ralstonia solanacearum[J].Chinese Journal of Biological Control.2002,18(4):185-187.
[6]何礼远,康耀卫.利用青枯菌无毒菌株和荧光假单胞菌诱导花生产生抗病性[J].植物保护学报,1990,17(2):113-116.doi:10.13802/j.cnki.zwbhxb.1990.02.005.HE L Y,KANG Y W.Induced resistance of peanut produced by avirulent strains of Ralstonia solanacearum and fluorescentPseudomonad[J].Acta phytophylacica sinica,1990,17(2):113-116.doi:10.13802/j.cnki.zwbhxb.1990.02.005.
[7]郭坚华,王玉菊,李瑾,任欣正.抑菌圈-定殖力双重测定法筛选青枯病生防细菌[J].植物病理学报,1996,26(1):49-54.DOI:10.13926/j.cnki.apps.1996.01.014.GUO J H,WANG Y J,LI J,REN X Z.Screen of biocontrol bacteria of plant wilt by inhibiting zones and root-colonizing capacity[J].Acta Phytophylacica Sinica,1996,26(1):49-54.
[8]杨合同,任欣正,王少杰,王建平,高强.蕃茄根土区系中拮抗性细菌的分布与青枯病发生的关系[J].生物防治通报,1994,10(4):162-165.doi:10.16409/j.cnki.2095-039x.1994.04.004.YANG H T,REN X Z,WANG S J,WANG J P,GAO Q.Study on distribution of antagonistic bacteria in root-soil system of tomato and ITS relation to occurrence of bacterial wilt[J].Chinese Journal of Biological Control,1994,10(4):162-165.doi:10.16409/j.cnki.2095-039x.1994.04.004.
[9]蒲小明,林壁润,郑奕雄,沈会芳.一株抗花生青枯病菌海洋放线菌的分类鉴定及其活性产物研究[J].广东农业科学,2011,38(16):62-65.doi:10.16768/j.issn.1004-874x.2011.16.039.PU X M,LIN B R,ZHENG Y X,SHEN H F.Studies on taxonomic identification of a marine actinomycete against Pseudomonas solanacearum and its active product[J].Guangdong Agricultural Sciences,2011,38(16):62-65.doi:10.16768/j.issn.10 04-874x.2011.16.039.
[10]樊胜南,喻国辉,陈燕红,黎永坚,陈川雁.枯草芽孢杆菌TR21可湿性粉剂对香蕉根际土壤微生物的影响[J].广东农业科学,2018,45(10):68-73.doi:10.16768/j.issn.1004-874X.2018.10.011.FAN S N,YU G H,CHEN Y H,LI Y J,CHEN C Y.Influence of Bacillus subtilis strain TR21 wettable powder on the banana rhizosphere microorganisms[J].Guangdong Agricultural Sciences,2018,45(10):68-73.doi:10.16768/j.issn.1004-874X.2018.10.011.
[11]游春平,刘凤权.植物病原细胞研究技术[M].北京:中国农业出版社,2011.YOU C P,LIU F Q.Plant Pathogenic Cell Research Technology[M].Beijing:China Agricultural Press,2011.
[12]张建华,任欣正,方中达.青枯假单胞杆菌产生细菌素的研究[J].南京农业大学学报,1987,10(3):58-63.ZHANG J H,REN X Z,FANG Z D.Studies on bacteriocin production of Pseudomonas solanacearum[J].Journal of Nanjing agricultural university,1987,10(3):58-63.
[13]KOUMOUTSI A,CHEN X H,HENNE A,LIESEGANG H,HITZEROTH G,FRANKE P.VATER J.BORRISS R.Structural and functional characterization of gene-clusters directing nonribosomal synthesis of bioactive cycliclipopeptides in Bacillus amyloliquefaciens strains FZB42[J].Journal of Bacteriology,2004,186(4):1084-1096.doi:10.1128/JB.186.4.1084-1096.2004.
[14]刘波.青枯雷尔氏菌的生物学及其生物防治[M].福州:福建省农科院农业生物资源研究所农业微生物研究中心,2003.LIU B.Biology of Ralstonia solanacearum and its biological control[M].Fuzhou:Agricultural Microbiology Research Center,Institute of Agricultural Biological Resources,Fujian Academy of Agricultural Sciences,2003.
[15]喻国辉,程萍,王燕鹂,陈远凤,陈燕红,黎永坚.枯草芽孢杆菌TR21田间防治巴西蕉枯萎病的效果[J].中国生物防治学报,2010,26(4):497-500.doi:10.16409/j.cnki.2095-039x.2010.04.020.YU G H,CHENG P,WANG Y L,CHEN Y F,CHEN Y H,LI Y J.Control effect of bacillus subtilis Strain TR21 on panama disease of banana brazil(Musa spp.)in fields[J].Chinese Journal of Biological Control,2010,26(4):497-500.doi:10.16409/j.cnki.2095-039x.2010.04.020.
[16]王成义,闫茂仓,陈少波,管敏鑫,单乐州,艾为明,谢起浪,蔡延马奔.基于16S rRNA和recA香鱼鳗利斯顿氏菌的分离鉴定[J].海洋通报,2010,29(1):84-90.WANG C Y,YAN M C,CHEN S B,GUAN M X,SHAN L Z,AIW M,XIE Q L,CAI Y B.Isolation and identification of Listonella anguillarum from Plecoglossus altivelis based on 16S rRNA and recA[J].Marine science bulletin,2010,29(1):84-90.
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