芽孢杆菌BS-6基于全基因组数据的分类鉴定及拮抗 能力分析

doi:10.13560/j.cnki.biotech.bull.1985.2019-0394

生物技术通报 ›› 2019, Vol. 35 ›› Issue (10): 111-118.doi: 10.13560/j.cnki.biotech.bull.1985.2019-0394

芽孢杆菌BS-6基于全基因组数据的分类鉴定及拮抗能力分析

戚家明^{1, 2, 3, 4}, 孙杉杉^{1, 2, 3}, 张东旭^{1, 2, 3}, 徐志文^{1, 2, 3, 4}, 徐延平^{1, 2, 3, 4}

1. 河北省农业生物技术工程技术研究中心,秦皇岛 066004;
2. 河北省农业厅植物营养与海洋功能肥料重点实验室,秦皇岛 066004;
3. 领先生物农业股份有限公司,秦皇岛 066004;
4. 秦皇岛领先康地农业技术有限公司,秦皇岛 066400

收稿日期:2019-05-08 出版日期:2019-10-26 发布日期:2019-09-30
作者简介:戚家明,男,硕士,研究方向：微生物发酵;E-mail：qi.jiaming@leadst.cn
基金资助:
河北省产业创新创业团队（169A76114H）,高效微生物肥料关键技术研究与应用（201801B027）,畜禽粪便纳米膜好氧发酵堆肥技术推广与应用（HBCT2018110401）

Identification and Biocontrol Activity Analysis of Bacillus sp. BS-6 Based on Genome-wide Data

QI Jia-ming^{1, 2, 3, 4}, SUN Shan-shan^{1, 2, 3}, ZHANG Dong-xu^{1, 2, 3}, XU Zhi-wen^{1, 2, 3, 4}, XU Yan-ping^{1, 2, 3, 4}

1. Hebei Agricultural Biotechnology Engineering Technology Research Center,Qinhuangdao 066004;
2. Key Laboratory of Plant Nutrition and Marine Functional Fertilizer of Hebei Provincial Agricultural Department,Qinhuangdao 066004;
3. Leading Bio-agricultural Co. Ltd,Qinhuangdao 066004;
4. Qinhuangdao Leading Comdee Agricultural Technology Co. Ltd,Qinhuangdao 066400

Received:2019-05-08 Published:2019-10-26 Online:2019-09-30

摘要/Abstract

摘要： 基于全基因组数据,确定芽孢杆菌BS-6的分类地位,验证其对植物病原菌的拮抗作用以及挖掘其潜在的生防功能。通过全基因组中16S rRNA及gyrA基因序列的系统发育分析及基因组分析方法确定芽孢杆菌BS-6的分类地位,通过平板对峙方法研究其对马铃薯枯萎病菌、玉米纹枯病菌和苹果轮纹病菌的拮抗能力;利用antiSMASH软件分析和预测菌株BS-6抑菌物质的相关基因并挖掘其生防潜力。基于16S rRNA及gyrA基因序列的系统发育分析及基因组分析结果,BS-6被鉴定为枯草芽孢杆菌,同时发现BS-6基因组中存在7种芽孢杆菌属重要或特有的次生代谢产物基因簇,4种未知功能的次生代谢产物基因簇,具有良好的抑制真菌生长的能力。枯草芽孢杆菌BS-6具有较强的拮抗植物病原菌的能力及良好的农业应用前景。

关键词: 枯草芽孢杆菌, 拮抗能力, 基因组, 基因簇

Abstract: Based on the genome-wide data,the taxonomic status of Bacillus sp. BS-6 isolated from soil was clarified. The antagonistic effect of Bacillus sp. BS-6 on multiple plant pathogens was validated to uncover its potential biocontrol functions. Phylogenetic trees were constructed by 16S rRNA and gyrA gene sequences analysis to identify Bacillus sp. BS-6. Moreover,whole-genome sequencing analysis was also employed to determine the taxonomic status of strain BS-6. Plate isolate-antagonistic experiments were applied to explore the antagonistic effect of strain BS-6 on Fusarium oxysporum,Rhizoctonia solani and Botryospuaeria berengeriana. antiSMASH was used to analyze and predict the antibiotic genes of strain BS-6 among secondary metabolite biosynthesis gene clusters and to explore its biocontrol potential. According to the results of phylogenetic trees,average nucleotide identity and digital DNA-DNA hybridization,BS-6 was characterized as Bacillus subtilis. The BS-6,with 7 important or unique secondary metabolite gene clusters of Bacillus and 4 unknown function secondary metabolite gene clusters in genome,exhibited a fine ability of inhibiting the growth of fungi. Bacillus subtilis BS-6 has strong antagonistic ability against plant pathogens and promising application prospects in agriculture.

Key words: Bacillus subtilis, antagonistic ability, genome, gene clusters

戚家明, 孙杉杉, 张东旭, 徐志文, 徐延平. 芽孢杆菌BS-6基于全基因组数据的分类鉴定及拮抗能力分析[J]. 生物技术通报, 2019, 35(10): 111-118.

QI Jia-ming, SUN Shan-shan, ZHANG Dong-xu, XU Zhi-wen, XU Yan-ping. Identification and Biocontrol Activity Analysis of Bacillus sp. BS-6 Based on Genome-wide Data[J]. Biotechnology Bulletin, 2019, 35(10): 111-118.

参考文献

[1] Meinken C, Blencke HM, Ludwig H, et al.Expression of the glycolytic gapA operon in Bacillus subtilis:differential syntheses of proteins encoded by the operon[J]. Microbiology, 2003, 149(3):751-761.
[2] Mukherjee S, Das P, Sen R.Towards commercial production of microbial surfactants[J]. Trends in Biotechnology, 2006, 24(11):509-515.
[3] 赵朋超, 王建华, 权春善, 等. 枯草芽孢杆菌抗菌肽生物合成的研究进展[J]. 中国生物工程杂志, 2010, 30(10):108-113.
[4] Leclère V, Béchet M, Adam A, et al.Mycosubtilin overproduction by Bacillus subtilis BBG100 enhances the organism's antagonistic and biocontrol activities[J]. Appl Environ Microbiol. , 2005, 71(8):4577-4584.
[5] Stein T.Bacillus subtilis antibiotics:structures, syntheses and specific functions[J]. Molecular Microbiology, 2005, 56(4):845-857.
[6] Nguyen ATV, Nguyen DV, Tran MT, et al.Isolation and characterization of Bacillus subtilis CH16 strain from chicken gastrointestinal tracts for use as a feed supplement to promote weight gain in broilers[J]. Letters in Applied Microbiology, 2015, 60(6):580-588.
[7] Kim B, Lee S, Lyu M, et al.Identification of mycobacterial species by comparative sequence analysis of the RNA polymerase gene(rpoB)[J]. Journal of Clinical Microbiology, 1999, 37(6):1714-1720.
[8] Wang LT, Lee FL, Tai CJ, et al.Comparison of gyrB gene sequences, 16S rRNA gene sequences and DNA-DNA hybridization in the Bacillus subtilis group[J]. International Journal of Systematic and Evolutionary Microbiology, 2007, 57(8):1846-1850.
[9] Rosselló-Mora R.Updating prokaryotic taxonomy[J]. J Bacteriol, 2005, 187(18):6255-6257.
[10] Richter M, Rosselló-Móra R.Shifting the genomic gold standard for the prokaryotic species definition[J]. Proceedings of the National Academy of Sciences of the United States of America, 2009, 106(45):19126-19131.
[11] Konstantinidis KT, Tiedje J M.Genomic insights that advance the species definition for prokaryotes[J]. Proceedings of the National Academy of Sciences of the United States of America, 2005, 102(7):2567-2572.
[12] McCarthy BJ, Bolton ET. An approach to the measurement of genetic relatedness among organisms[J]. Proceedings of the National Academy of Sciences of the United States of America, 1963, 50(1):156.
[13] Adamek M, Spohn M, Stegmann E, et al.Antibiotics[M]. New York:Humana Press, 2017.
[14] Richter M, Rosselló-Móra R, Oliver Glöckner F, et al.JSpeciesWS: a web server for prokaryotic species circumscription based on pairwise genome comparison[J]. Bioinformatics, 2016, 32(6):929-931.
[15] Meier-Kolthoff JP, Auch AF, Klenk HP, et al.Genome sequence-based species delimitation with confidence intervals and improved distance functions[J]. BMC Bioinformatics, 2013, 14(1):60-74.
[16] Weber T, Blin K, Duddela S, et al.antiSMASH 3. 0-a comprehen-sive resource for the genome mining of biosynthetic gene clusters[J]. Nucleic Acids Research, 2015, 43(W1):W237-W243.
[17] 冯兴, 潘康成, 张顺泉. 一株益生芽孢杆菌Pab02的16S rDNA测序鉴定[J]. 中国饲料, 2008(18):4-7.
[18] Fukushima M, Kakinuma K, Kawaguchi R.Phylogenetic analysis of Salmonella, Shigella, and Escherichia coli strains on the basis of the gyrB gene sequence[J]. Journal of Clinical Microbiology, 2002, 40(8):2779-2785.
[19] Nagao JI.Properties and applications of lantibiotics, a class of bacteriocins produced by Gram-positive bacteria[J]. Journal of Oral Biosciences, 2009, 51(3):158-164.
[20] Spieβ T, Korn SM, Kötter P, et al.Autoinduction specificities of the lantibiotics subtilin and nisin[J]. Appl Environ Microbiol. , 2015, 81(22):7914-7923.
[21] Fuchs SW, Jaskolla TW, Bochmann S, et al.Entianin, a novel subtilin-like lantibiotic from Bacillus subtilis subsp. spizizenii DSM 15029T with high antimicrobial activity[J]. Appl Environ Microbiol, 2011, 77(5):1698-1707.
[22] Babasaki K, Takao T, Shimonishi Y, et al.Subtilosin A, a new antibiotic peptide produced by Bacillus subtilis 168:isolation, structural analysis, and biogenesis[J]. The Journal of Biochemistry, 1985, 98(3):585-603.
[23] Liu Q, Gao G, Xu H, et al.Identification of the bacteriocin subtilosin A and loss of purL results in its high-level production in Bacillus amyloliquefaciens[J]. Research in Microbiology, 2012, 163(6-7):470-478.
[24] Jasim B, Sreelakshmi KS, Mathew J, et al.Surfactin, iturin, and fengycin biosynthesis by endophytic Bacillus sp. from Bacopa monnieri[J]. Microbial Ecology, 2016, 72(1):106-119.
[25] Singh P, Cameotra SS.Potential applications of microbial surfactants in biomedical sciences[J]. Trends in Biotechnology, 2004, 22(3):142-146.
[26] Jain RM, Mody K, Joshi N, et al.Effect of unconventional carbon sources on biosurfactant production and its application in bioremediation[J]. International Journal of Biological Macromolecules, 2013, 62:52-58.
[27] Al-Ajlani MM, Sheikh MA, Ahmad Z, et al.Production of surfactin from Bacillus subtilis MZ-7 grown on pharmamedia commercial medium[J]. Microbial Cell Factories, 2007, 6(1):17-25.
[28] Chen WC, Juang RS, Wei YH.Applications of a lipopeptide biosurfactant, surfactin, produced by microorganisms[J]. Biochemical Engineering Journal, 2015, 103:158-169.
[29] Banat IM, De Rienzo MAD, Quinn GA.Microbial biofilms:biosurfactants as antibiofilm agents[J]. Applied Microbiology and Biotechnology, 2014, 98(24):9915-9929.
[30] Mnif I, Hammami I, Triki MA, et al.Antifungal efficiency of a lipopeptide biosurfactant derived from Bacillus subtilis SPB1 versus the phytopathogenic fungus, Fusarium solani[J]. Environmental Science and Pollution Research, 2015, 22(22):18137-18147.
[31] Hotta K, Kim CY, Fox DT, et al.Siderophore-mediated iron acquisition in Bacillus anthracis and related strains[J]. Microbiology, 2010, 156(7):1918-1925.
[32] Müller S, Strack SN, Hoefler BC, et al.Bacillaene and sporulation protect Bacillus subtilis from predation by Myxococcus xanthus[J]. Appl Environ Microbiol, 2014, 80(18):5603-5610.
[33] Butcher RA, Schroeder FC, Fischbach MA, et al.The identification of bacillaene, the product of the PksX megacomplex in Bacillus subtilis[J]. Proceedings of the National Academy of Sciences, 2007, 104(5):1506-1509.
[34] Besson F, Michel G.Action of mycosubtilin, an antifungal antibiotic of Bacillus subtilis, on the cell membrane of Saccharomyces cerevisiae[J]. Microbios, 1989, 59(239):113-121.
[35] Thimon L, Peypoux F, Dana Maget R, et al.Interactions of bioactive lipopeptides, iturin A and surfactin from Bacillus subtilis[J]. Biotechnology and Applied Biochemistry, 1992, 16(2):144-151.
[36] Borisova SA, Circello BT, Zhang JK, et al.Biosynthesis of rhizocticins, antifungal phosphonate oligopeptides produced by Bacillus subtilis ATCC6633[J]. Chemistry & Biology, 2010, 17(1):28-37.
[37] Hourdou ML, Besson F.Surfactine et antibiotiques ituriniques:structure, propriétés et biosynthèse des lipopeptides de Bacillus subtilis[J]. Regard sur la biochimie, 1994(1):35-42.
[38] Besson F, Peypoux F, Michel G, et al.Antifungal activity upon Saccharomyces cerevisiae of iturin A, mycosubtilin, bacillomycin L and of their derivatives;inhibition of this antifungal activity by lipid antagonists[J]. The Journal of Antibiotics, 1979, 32(8):828-833.

芽孢杆菌BS-6基于全基因组数据的分类鉴定及拮抗能力分析

Identification and Biocontrol Activity Analysis of Bacillus sp. BS-6 Based on Genome-wide Data

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芽孢杆菌BS-6基于全基因组数据的分类鉴定及拮抗 能力分析

Identification and Biocontrol Activity Analysis of Bacillus sp. BS-6 Based on Genome-wide Data

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芽孢杆菌BS-6基于全基因组数据的分类鉴定及拮抗能力分析