枯草芽胞杆菌GB519抗菌蛋白的理化性质及生防效果

doi:10.16409/j.cnki.2095-039x.2020.05.024

中国生物防治学报 ›› 2020, Vol. 36 ›› Issue (5): 778-785.DOI: 10.16409/j.cnki.2095-039x.2020.05.024

枯草芽胞杆菌GB519抗菌蛋白的理化性质及生防效果

朱峰¹, 王继春¹, 田成丽¹, 祁山颜^1,2, 王东元², 任金平¹, 王义山³

1. 吉林省农业科学院植物保护研究所/农业部东北作物有害生物综合治理重点实验室, 公主岭 136100;
2. 吉林农业大学植物保护学院, 长春 130118;
3. 双辽市东明镇林业站, 双辽 136414

收稿日期:2020-02-14 出版日期:2020-10-08 发布日期:2020-11-20
通讯作者: 王继春,博士,副研究员,E-mail:wangjichun@cjaas.com。
作者简介:朱峰,博士,助理研究员,E-mail:zhufeng0726@163.com
基金资助:
吉林省农业科技创新工程（CXGC2017TD010）；吉林省科技厅优秀青年人才基金（20190103131JH）；吉林省农业科学院博士后基金（188320）

Physicochemical Characteristics and Biocontrol Efficacy of Antagonistic Protein from Bacillus subtilis GB519

ZHU Feng¹, WANG Jichun¹, TIAN Chengli¹, QI Shanyan^1,2, WANG Dongyuan², REN Jinping¹, Wang Yishan³

1. Key Laboratory of Integrated Pest Management on Crops in Northeast, Ministry of Agriculture Institute of Plant Protection/Jilin Academy of Agricultural Sciences, Gongzhuling 136100, China;
2. College of Plant Protection, Jinlin Agricultural University, Changchun 130118, China;
3. Forest Station of Dongming Town, Shuangliao 136414, China

Received:2020-02-14 Online:2020-10-08 Published:2020-11-20

摘要/Abstract

摘要： 枯草芽胞杆菌GB519是从土壤中分离对稻瘟病菌具有显著抑菌活性的菌株。为了明确菌株GB519抗菌粗蛋白的理化性质，分析了光照、紫外线、温度、pH和酶（蛋白酶K、胰蛋白酶和胃蛋白酶）对抑菌活性的影响。结果表明，抗菌蛋白对光照、紫外线和3种酶作用稳定；40℃~100℃处理1 h抑菌活性稳定，121℃处理30 min活性仅降低了7.4%，具有较好的耐高温特性；在pH 2.0~10.0活性稳定，但pH 11.0~12.0活性有所下降，分别降低了14.9%和18.1%。抗菌蛋白对多种植物病原菌具有抑菌作用，其中对茄子菌核病菌的抑菌活性为72.7%。在温室条件下对水稻叶瘟的防效达到80.1%。因此，菌株GB519可作为防控稻瘟病生防潜力菌进行开发。

关键词: 枯草芽胞杆菌GB519, 抑菌蛋白, 理化性质, 水稻稻瘟病

Abstract: The strain GB519 of Bacillus subtilis was isolated from the soil and showed significant antimicrobial activity against rice blast caused by Magnaporthe oryzae. In order to clarify the physicochemical characteristics of antagonistic crude protein of strain GB519, the effects of illumination, UV, temperature, pH and enzymes (proteinase K, trypsase and pepsase) on antimicrobial activity were analyzed. The results showed that antagonistic protein was stable in illumination, UV and three kinds of enzymes; Antimicrobial activity remained stable at 40-100℃ for 1 h and decreased by only 7.4% at 121℃ for 30 min, showing good resistance to high temperature; Antimicrobial activity remained stable from pH 2.0 to 10.0, and declined by 14.9% and 18.1% at pH 11.0 and pH 12.0, respectively. Antagonistic protein had antagonistic effect on multiple crop pathogens, especially among which the antimicrobial activity against Sclerotinia sclerotiorum was 72.7%. Under greenhouse conditions, the biocontrol efficacy against leaf blast reached 80.1%. Therefore, the B. subtilis GB519 could be developed as biocontrol potential bacteria for the control of rice blast.

Key words: Bacillus subtilis GB519, antagonistic protein, physicochemical properties, Magnaporthe oryzae

中图分类号:

S476

朱峰, 王继春, 田成丽, 祁山颜, 王东元, 任金平, 王义山. 枯草芽胞杆菌GB519抗菌蛋白的理化性质及生防效果[J]. 中国生物防治学报, 2020, 36(5): 778-785.

ZHU Feng, WANG Jichun, TIAN Chengli, QI Shanyan, WANG Dongyuan, REN Jinping, Wang Yishan. Physicochemical Characteristics and Biocontrol Efficacy of Antagonistic Protein from Bacillus subtilis GB519[J]. Chinese Journal Of Biological Control, 2020, 36(5): 778-785.

参考文献

[1] Yasin F D, Kae Y, Gulay D. Septin-mediated plant cell invasion by the rice blast fungus, Magnaporthe oryzae[J]. Science, 2012, 336(6088):1590-1595.
[2] Miah G, Rafii M Y, Ismail M R, et al. Blast resistance in rice:a review of conventional breeding to molecular approaches[J]. Molecular Biology Reports, 2012, 40:2369-2388.
[3] Todorova S, Kozhuharova L. Characteristics and antimicrobial activity of Bacillus subtilis strains isolated from soil[J]. World Journal of Microbiology & Biotechnology, 2010, 26:1207-1216.
[4] 张丽霞, 李荣禧, 王琦, 等. 枯草芽孢杆菌发酵培养基的优化[J]. 中国生物防治, 2006, 22(S):82-88.
[5] Tokpah D P, Li H W, Wang L Y, et al. An assessment system for screening effective bacteria as biological control agents against Magnaporthe grisea on rice[J]. Biological Control, 2016, 103(1):21-29.
[6] Ghasemi S, Ahmadian G, Sadeghi M, et al. First report of a bifunctional chitinase/lysozyme produced by Bacillus pumilus SG2[J]. Enzyme and Microbial Technology, 2011, 48(3):225-231.
[7] Huang X, Yong X, Zhang R, et al. The supernatant of Bacillus pumilus SQR-N43 has antifungal activity towards Rhizoctonia solani[J]. Journal of Basic Microbiology, 2013, 53(8):657-663.
[8] 沙月霞, 王琦, 李燕. 稻瘟病生防芽胞杆菌的筛选及防治效果[J]. 中国生物防治学报, 2016, 32(4):474-484.
[9] Wang N N, Yan X, Gao X N, et al. Purification and characterization of a potential antifungal protein from Bacillus subtilis E1R-J against Valsa mali[J]. World Journal of Microbiology and Biotechnology, 2016, 32:63.
[10] 胡晓丹, 王建伟, 李孝敬, 等. 赤霉病菌拮抗菌Bacillus subtilis AF0907抗菌物质研究[J]. 中国生物防治学报, 2015, 31(3):378-385.
[11] 于杰, 张荣意, 谭志琼, 等. 枯草芽孢杆菌B25抗真菌作用及抗菌蛋白的分离纯化[J]. 基因组学及应用生物学, 2016, 35(3):629-634.
[12] Rao Q, Guo W B, Chen X H. Identification and characterization of an antifungal protein, AfAFP_R9, produced by marine-derived Aspergillus fumigatus R9[J]. Journal of Microbiology and Biotechnology, 2015, 25(5):620-628.
[13] 王法国, 许婷婷, 张荣胜, 等. 解淀粉芽胞杆菌干悬浮剂对稻瘟病的防治效果及安全性评价[J]. 中国生物防治学报, 2017, 33(2):241-247.
[14] Chen H, Wang L, Su C X, et al. Isolation and characterization of lipopeptide antibiotics produced by Bacillus subtilis[J]. Letters in Applied Microbiology, 2008, 47(3):180-186.
[15] Prabavathy V R, Mathivanan N, Murugesan K. Control of blast and sheath blight diseases of rice using antifungal metabolites produced by Streptomyces sp. PM5[J]. Biological Control, 2006, 39(3):313-319.
[16] Spence C, Alff E, Johnson C, et al. Natural rice rhizospheric microbes suppress rice blast infections[J]. BMC Plant Biology, 2014, 14(1):130.
[17] Meng X K, Yu J J, Yu M N, et al. Dry flowable formulations of antagonistic Bacillus subtilis strain T429 by spray drying to control rice blast disease[J]. Biological Control, 2015, 85(1):46-51.
[18] Fan H Y, Ru J J, Zhang Y Y, et al. Fengycin produced by Bacillus subtilis 9407 plays a major role in the biocontrol of apple ring rot disease[J]. Microbiological Research, 2017, 199:89-97.
[19] 黄娜, 周莉质, 费丹, 等. 枯草芽胞杆菌胞外抗菌蛋白的初步分离及性质的研究[J]. 植物保护, 2015, 41(6):49-54.
[20] 刘静, 王军, 姚建铭, 等. 枯草芽孢杆菌JA抗菌物特性的研究及抗菌肽的分离纯化[J]. 微生物学报, 2004, 44(4):511-514.
[21] Zhao J, Wu Y X, Ho H H, et al. PBT1, a novel antimicrobial protein from the biocontrol agent Bacillus subtilis XF-1 against Plasmodiophora brassicae[J]. European Journal of Plant Pathology, 2016, 145(3):583-590.
[22] krid S, Rhouma A, Mogou I, et al. Pseudomonas savastanoi endophytic bacteria in olive tree knots and antagonistic potential of strains of Pseudomonas fluorescens and Bacillus subtilis[J]. Journal of Plant Pathology, 2010, 92(2):335-341.
[23] Li J, Yang Q, Zhao L H, et al. Purification and characterization of a novel antifungal protein from Bacillus subtilis strain B29[J]. Journal of Zhejiang University Science B, 2009, 10(4):264-272.
[24] 裴韬, 任大明, 石皎. 小麦赤霉病拮抗菌P72抗菌物质的分离纯化和性质研究[J]. 安徽农业科学, 2009, 37(6):2576-2577.
[25] 沈锦玉, 尹文林, 曹铮, 等. 枯草芽孢杆菌B115抗菌蛋白的分离纯化及部分性质[J]. 水生生物学报, 2005, 29(6):689-693.
[26] 童有仁, 马志超, 陈卫良, 等. 枯草芽孢杆菌B034拮抗蛋白的分离纯化及特性分析[J]. 微生物学报, 1999, 39(4):339-343.
[27] 陈志谊, 许志刚, 陆凡, 等. 拮抗细菌B-916培养液对水稻纹枯病的抗生活性及其抗菌物质的研究[J]. 江苏农业科学, 2000, 16(3):148-152.
[28] Magali D, Michel P. Effect of fengycin, a lipopeptide produced by Bacillus subtilis, on model biomembranes[J]. Biophysical Journal, 2008, 94:2667-2679.
[29] Stein T. Bacillus subtilis antibiotics:structure, syntheses and specific functions[J]. Molecular Microbiology, 2005, 56(4):845-857.
[30] Carrillo C, Teruel J A, Aranda F J, et al. Molecular mechanism of membrane permeabilization by the peptide antibiotic surfactin[J]. Biochimica et Biophysica Acta-Biomembranes, 2003, 1611(1-2):91-97.
[31] Ongena M, Jacques P. Bacillus lipopeptides versatile weapons for plant disease biocontrol[J]. Trends in Microbiology, 2008, 16(1):115-125.
[32] 穆常青, 刘雪, 陆庆光, 等. 枯草芽胞杆菌B-332菌株对稻瘟病的防治效果及定殖作用[J]. 植物保护学报, 2007, 34(2):123-128.

枯草芽胞杆菌GB519抗菌蛋白的理化性质及生防效果

Physicochemical Characteristics and Biocontrol Efficacy of Antagonistic Protein from Bacillus subtilis GB519

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 6

编辑推荐

Metrics

本文评价

联系我们

[1]	胡飞, 胡本进, 周子燕, 徐丽娜, 苏卫华, 苏贤岩. 解淀粉芽胞杆菌JDF-6液态发酵条件优化及抑菌蛋白的分析[J]. 中国生物防治学报, 2017, 33(4): 561-570.
[2]	王祺, 张一名, 赵君, 景岚, 周洪友. 枯草芽孢杆菌S-16抑菌活性物质的理化性质及培养条件的研究[J]. , 2015, 31(3): 439-444.
[3]	余莎1 , 何红2 , 詹儒林2 , 马凌燕1 , 李琳1 . 红树内生细菌AiL3抗菌蛋白的纯化及其防治芒果炭疽病机理研究[J]. , 2013, 29(1): 104-109.
[4]	张晓云, 李宝庆, 郭庆港, 鹿秀云, 李社增, 马平. 枯草芽孢杆菌CAB-1抑菌蛋白对黄瓜白粉病的防治作用[J]. , 2012, 28(3): 375-380.
[5]	魏松红, 付丹妮, 逄若霖, 张丽莹, 王黎. 除草活性放线菌DN5发酵条件及发酵产物理化性质的研究[J]. , 2011, 27(2): 246-253.
[6]	穆常青1,2 , 潘玮1,2 , 陆庆光2 , 蒋细良1 , 朱昌雄1 . 枯草芽孢杆菌对稻瘟病的防治效果评价及机制初探[J]. , 2006, 22(2): 158-160.