一株番茄青枯病拮抗细菌JK19的鉴定及其抑菌活性检测

doi:10.6048/j.issn.1001-4330.2021.07.020

新疆农业科学 ›› 2021, Vol. 58 ›› Issue (7): 1355-1364.DOI: 10.6048/j.issn.1001-4330.2021.07.020

• 园艺特产·微生物·草业 • 上一篇下一篇

一株番茄青枯病拮抗细菌JK19的鉴定及其抑菌活性检测

黄伟¹, 宋博², 张丽娟¹, 王博¹, 王玮¹

1.新疆农业科学院微生物应用研究所/新疆特殊环境微生物实验室,乌鲁木齐 830091;
2.新疆农业科学院植物保护研究所/农业部西北荒漠绿洲作物有害生物防治综合治理重点实验室/农业部库尔勒作物有害生物科学观测试验站,乌鲁木齐 830091

收稿日期:2020-05-13 发布日期:2021-07-27
通信作者: 王玮(1973-),女,新疆人,研究员,研究方向为微生物资源,(E-mail)mypony926@163.com
作者简介:黄伟(1991-),男,新疆阿图什人,助理研究员,研究方向为微生物资源,(E-mail)1428775150@qq.com
基金资助:
自治区重点实验室开放课题“基于OSMAC策略的耐辐射链霉菌A144农用天然产物挖掘与研究”(2020D04032);新疆农业科学院青年基金“青枯病拮抗菌的筛选、鉴定及其活性物质的初步分析”(xjnkq-2018010);国家生物农药工程技术研究中心及农业部农业微生物资源利用华中区域科学观测试验站联合开放基金”(JF-NBCOES-1802);新疆农业科学院青年科技骨干创新能力培养项目“一株高效杀线虫芽孢杆菌活性物质提取及作用机制研究”(xjnkq-2019019)

Identification of Bacterial Wilt Antagonistic Bacteria JK19 and Detection of Its Antibacterial Activity

HUANG Wei¹, SONG Bo², ZHANG Lijuan¹, WANG Bo¹, WANG Wei¹

1. Xinjiang Laboratory of Special Environmental Microbiology /Institute of Applied Microbiology, Xinjiang Academy of Agricultural Sciences, Urumqi 830091, China;
2. Key Laboratory of Integrated Crop Pest Management in Northwestern Oasis of China,MOARA/ Scientific Observing and Experimental Station of Crop Pests of MOARA in Korla / Institute of Plant Protection,Xinjiang Academy of Agricultural Sciences,Urumqi 830091,China

Received:2020-05-13 Published:2021-07-27
Correspondence author: WANG Wei (1973-), female, born in Xinjiang Province, research interest in microbial resources, (E-mail)mypony926@163.com
Supported by:
Youth Fund of Xinjiang Academy of Agricultural Sciences "Screening, Identification and Preliminary Analysis of Antibacterial Substances of Prunus solani"(xjnkq-2018010); Joint Open Fund for the Utilization of Agricultural Microbial Resources from the Central China Regional Scientific Observation and Test Station of National Research Centre for Biopesticide Engineering Technology and MOARR (JF-NBCOES-1802); Youth Science and Technology Backbone Innovation Ability Training Project of Xinjiang Academy of Agricultural Sciences "Study on the Extraction and Mechanism of Active Substance from a High Efficiency Bacillus Nematode"(xjnkq-2019019)

摘要/Abstract

摘要： 【目的】研究拮抗细菌JK19的分类学地位,检测其抑菌活性,分析其可能的活性成分。为脂肽提取和生防制剂的研制提供了理论依据。【方法】采用形态学、生理生化特征及16S rDNA序列分析方法,鉴定菌株;采用平板对峙法检测菌株对8种植物病原菌的拮抗作用;采用PCR扩增方法检测基因组中的脂肽类抗生素合成及促生作用相关的基因。【结果】鉴定JK19为芽孢杆菌,JK19与贝莱斯芽孢杆菌、暹罗芽孢杆菌、解淀粉芽孢杆菌同源关系比较近。菌株JK19对多种植物病原菌具有明显的抑制作用,抑制效果分别为香梨黑斑病 66.67%、核桃腐烂病100%、马铃薯早疫病73.33%、西瓜枯萎病 60.71%、水稻恶苗病67.86%、棉花枯萎病 76.92%、西瓜蔓枯病100%。PCR反应检测JK19基因组中有参与脂肽类抗生素合成相关的基因 ituA、ituD、bamC、fenB、fenD、srfAB;参与生长素IAA合成的基因ysnE;促生作用相关的基因yndJ。【结论】拮抗细菌JK19为芽孢杆菌属细菌,对番茄青枯病病原菌等多种植物病原菌均有明显的抑制作用,其基因组中含有脂肽类和促生作用相关基因。

关键词: 芽孢杆菌; 抑菌谱; 脂肽类抗生素

Abstract: 【Objective】 To clarify the taxonomic status of antagonistic bacterium JK19, detect its antibacterial activity, and analyze its possible active components. The results provide theoretical basis for the extraction of lipopeptide and the development of biological control agents .【Methods】 The strain was identified by morphological, physiological and biochemical characteristics and 16S rDNA sequence analysis. The antagonistic effect of the strain against 8 plant pathogens was detected by plate confrontation method. PCR amplification was used to detect the genes related to the synthesis and probiotics of lipopeptide antibiotics in the genome.【Results】 JK19 was identified as bacillus by morphological, physiological and biochemical characteristics, and 16S rDNA sequences of JK19 were compared to construct a developmental tree. It was found that JK19 had close homologous relations with Bacillus velezensis, Bacillussiamensis and Bacillus amylobacillus. Strain JK19 had significant inhibition effect on various plant pathogens, and the inhibition effect was 66.67% for black spot of fragrant pear, 100% for walnut rot, 73.33% for potato early blight, 60.71% for watermelon blight, 67.86% for rice blight, 76.92% for cotton blight and 100% for watermelon vine blight, respectively. PCR reaction detected the genes involved in the synthesis of lipopeptide antibiotics in JK19 genome: ituA, ituD, bamC, fenB, fenD, srfAB, YsnE, a gene involved in the synthesis of auxin IAA and geneyndJassociated with probiotics.【Conclusion】 Antagonistic bacterium JK19 is abacillussp., which has obvious inhibitory effect on various plant pathogenic bacteria such as tomato bacterial wilt, and its genome contains lipopeptide and genes related to probiotics.

Key words: Bacillus sp.; inhibition spectrum; lipopeptide antibiotics

中图分类号:

黄伟, 宋博, 张丽娟, 王博, 王玮. 一株番茄青枯病拮抗细菌JK19的鉴定及其抑菌活性检测[J]. 新疆农业科学, 2021, 58(7): 1355-1364.

HUANG Wei, SONG Bo, ZHANG Lijuan, WANG Bo, WANG Wei. Identification of Bacterial Wilt Antagonistic Bacteria JK19 and Detection of Its Antibacterial Activity[J]. Xinjiang Agricultural Sciences, 2021, 58(7): 1355-1364.

参考文献

[1]Chunyu LI, Weicong H U, Pan B, et al. Rhizobacterium Bacillus amyloliquefaciens Strain SQRT3-Mediated Induced Systemic Resistance Controls Bacterial Wilt of Tomato[J]. Pedosphere, 2017, 27(6).
[2]郑雪芳,刘波,朱育菁, 等.番茄青枯病生防芽胞杆菌的筛选与鉴定[J]. 中国生物防治学报,2016,32(5):657-665.
ZHENG Xuefang, LIU Bo, ZHU Yujing, et al. Screening and identification of bacillus control from bacterial wilt of tomato [J]. Chinese Journal of Biological Control and Prevention, 2016, 32 (5): 657-665.
[3]王艳,常润磊,周旭东.青枯病快速检测研究概况[J]. 桉树科技,2012,29(2):53-58.
WANG Yan, CHANG Runlei, ZHOU Xudong. Rapid detection of bacterial wilt [J]. Eucalyptus Technology, 2012, 29(2): 53-58.
[4]邓建良,刘红彦,刘玉霞, 等.解淀粉芽孢杆菌YN-1抑制植物病原真菌活性物质鉴定[J]. 植物病理学报,2010,40(2):202-209.
DENG Jianliang, LIU Hongyan, LIU Yuxia, et al. Identification of the antifungal substances from Bacillus amyloliquefaciens strain YN-1[J]. Acta Phytopathologica Sinica, 2010, 40(2):202-209.
[5]Joshi R, McSpadden Gardener BB. Identification and Characterization of Novel Genetic Markers Associated with Biological Control Activities in Bacillus subtilis[J]. Phytopathology. 2006, 96(2):145-154.
[6]Isabel Mora , Jordi Cabrefiga, Emilio Montesinos. Antimicrobialpeptide genes in Bacillus strain from plant environments [J]. International Microbiology, 2011, 14(4): 213-223.
[7]周小江. 2株海洋生境芽孢杆菌的抑菌促生长作用及有关控制基因[D]. 青岛:青岛科技大学,2015.
ZHOU Xiaojiang. Fungistasic and growth promoting activity of marine Bacillus atrains and amplified about the control gene fragments[D]. Qingdao: Qingdao University of Science and Technology, 2015.
[8]Sumpavapol P, Tongyonk L, Tanasupawat S, Chokesajjawatee N, Luxananil P, Visessanguan W. Bacillus siamensis sp. nov., isolated from salted crab (poo-khem) in Thailand [J]. International Journal of Systematic & Evolutionary Microbiology, 2010; 60:2364-2370.
[9]Ruiz-Garcia C, Bejar V, Martinez-Checa F, et al. Bacillus velezensis sp. nov., a surfactant-producing bacterium isolated from the river Velez in Malaga, southern Spain [J]. International Journal of Systematic and Evolutionary Microbiology, 2005; 55:191-195.
[10]刘海洋,王琦,王伟, 等.拮抗菌AL7的鉴定及其生防特性的初步研究[J]. 植物保护,2018,44(2):53-60.
LIU Haiyang, WANG Qi, WANG Wei, et al. Species identification and biocontrol characteristics of antagonistic bacterium AL7[J]. Plant Protection, 2018, 44(2):53-60.
[11]卢彩鸽,张殿朋,刘霆, 等.解淀粉芽孢杆菌MH71的生防活性及脂肽类抗生素基因检测[J]. 植物保护,2015,41(3):12-18.
LU Caige, ZHANG Dianpeng, LIU Ting, et al. Detection of the genes encoding lipopeptide antibiotics and biocontrol activity of Bacillus amyloliquefaciens MH71 [J]. Plant Protection, 2015, 41(3):12-18.
[12]Wang LT, Lee FL, Tai CJ, et al. Bacillus velezensis is a later heterotypic synonym of Bacillus amyloliquefaciens [J]. International Journal of Systematic and Evolutionary Microbiology, 2008,58:671-675.
[13]Dunlap CA, Kim SJ, Kwon SW, et al. Bacillus velezensis is not a later heterotypic synonym of Bacillus amyloliquefaciens; Bacillus methylotrophicus, Bacillus amyloliquefaciens subsp. plantarum and Bacillus oryzicola are later heterotypic synonyms of Bacillus velezensis based on phylogenomics [J]. Int J Syst Evol Microbiol, 2016; 66:1212-1217.
[14]Fan B, Blom J, Klenk H P, et al.Bacillusamyloliquefaciens,Bacillusvelezensis,and Bacillus siamensis form an “Operational Group B.amyloliquefaciens” within the B. subtilis species complex [J]. Frontiers in Microbiology, 2017,8:22.
[15]朱华珺,周瑚,任佐华, 等.枯草芽孢杆菌JN005胞外抗菌物质及对水稻叶瘟防治效果[J]. 中国水稻科学:2020,(1):1-15.
ZHU Huajun, ZHOU Hu, REN Zuohua, et al. Extracellular antimicrobial substances produced by Bacillus subtilis JN005 and its efficacy on rice leaf blast [J]. Chinese Journal of Rice Science, 2020,(1):1-15.
[16]柳皓月,金辉,曾黎明, 等.杀线虫芽孢杆菌发酵条件优化及大孔树脂筛选[J]. 应用生态学报,2020,31(7):2287-2292.
LIU Haoyue, JIN Hui, ZENG Liming, et al. Optimization of the fermentation conditions of Bacillus nematodes and screening of microporous adsorption resin [J]. Chinese Journal of Applied Ecology: 2020,31(7):2287-2292.
[17]何宗均,路垚,李峰,等.农用微生物菌剂在黄瓜和水稻种植中的应用[J]. 山西农业科学,2020,48(3):407-410.
HE Zongjun, LU Yao, LI Feng, et al. Application test of agricultural microbial agent in cucumber and rice cultivation [J]. Journal of Shanxi Agricultural Sciences, 2020, 48(3):407-410.
[18]李新宇,李磊,陈利达, 等.番茄匍柄霉叶斑病拮抗细菌的筛选与鉴定[J]. 园艺学报,2020,47(2):741-748.
LI Xinyu, LI Lei, CHEN Lida, et al. Screening and identification of antagonistic bacteria against gray leaf spot of tomato caused by Stephyliumsolani [J]. Acta Horticulturae Sinica, 2020,47(2):741-748.
[19]Ongena M, Jacques P. Bacillus lipopeptides: versatile weapons for plant disease biocontrol [J]. Trends Microbiol, 2008, 16(3):115-125.
[20]Fan H, Ru J, Zhang Y, et al. Fengycin produced by Bacillus subtilis 9407 plays a major role in the biocontrol of apple ring rot disease [J]. Microbiol Research, 2017, 199: 89-97.
[21]Lina María González-Jaramillo, Francisco José Aranda, José Antonio Teruel, et al. Antimycotic activity of fengycin C biosurfactant and its interaction with phosphatidylcholine model membranes [J]. Colloids & Surfaces B Biointerfaces, 2017, 156:114-122.
[22]Heerklotz H, Seelig J. Leakage and lysis of lipid membranes induced by the lipopeptidesurfactin[J]. Eur Biophys J, 2007,36(4-5): 305-314.
[23]Ongena M, Jourdan E, Adam A, et al. Surfactin and fengycinlipopeptides of Bacillus subtilis as elicitors of induced systemic resistance in plants [J]. Environmental Microbiology, 2007, 9(4):1084-1090.
[24]Liu X, Ren B, Chen M, et al. Production and characterization of a group of bioemulsifiers from the marine Bacillus velezensis strain H3[J]. Applied Microbiology & Biotechnology, 2010, 87(5):1881-1893.
[25]金清,张蕾,蒋秋悦, 等. Bacillus velenzensis S3-1基因组中表面活性素、伊枯草菌素和丰原素合成基因簇的定位与分析[J]. 上海师范大学学报(自然科学版),2018,47(4):451-457.
JIN Qing, ZHANG Lei, JIANG Qiuyue, et al. Location and analysis of synthetic gene cluster of Bacillus velezensis S3-1 insurfactin, iturin and fengucin [J]. Journal of Shanghai Normal University(Natural Science Ed.), 2018, 47(4):451-457.
[26]朱弘元,康健,范昕,等.解淀粉芽孢杆菌B15产脂肽的分离鉴定及抑菌机理[J]. 江苏农业科学,2016,44(5):186-189.
ZHU Hongyuan, KANG Jian, FAN Xin, et al. Isolation and identification of lipopeptide from Bacillus amylobacillus B15 and its antibacterial mechanism [J]. Jiangsu Agricultural Sciences, 2016, 44(5):186-189.

一株番茄青枯病拮抗细菌JK19的鉴定及其抑菌活性检测

Identification of Bacterial Wilt Antagonistic Bacteria JK19 and Detection of Its Antibacterial Activity

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