Optimization of Fermentation Conditions of Biocontrol Bacteria HMQ20YJ04 and Evaluation of Its Effect on Botrytis cinerea

doi:10.16409/j.cnki.2095-039x.2023.02.057

Abstract

Abstract: Strain HMQ20YJ04 is an antagonistic bacterium isolated and screened from saline-alkali soil in Minqin, Gansu Province, which has strong antifungal activity against various plant pathogenic fungi. In order to improve the control effect of fermentation liquid, the number of viable bacteria in fermentation liquid was used as the detection index, the fermentation medium components and shake flask fermentation conditions were screened and optimized by single-factor test, orthogonal test, Plackett-Burman test and response surface method. The results showed that the optimal composition of fermentation medium were soluble starch 5 g/L, yeast extract 10 g/L, MgSO₄‧7H₂O 5 g/L and NaCl 5 g/L and the optimal fermentation conditions were rotation speed 200 r/min, inoculation amount 5%, temperature 32 ℃, cultivation time 60 h and initial pH 6.5. The results of pot experiments in greenhouse showed that the fermentation liquid of strain HMQ20YJ04 had a good control effect on tomato gray mold, with the control effect of 63.03%. The above results provided a basis for the research and development of Bacillus subtilis strain HMQ20YJ04 agent and its application in biological control of tomato gray mold.

Key words: Botrytis cinerea, Bacillus subtilis, orthogonal experiment design, Plackett-Burman test, response surface method

CLC Number:

S476

ZHANG Jinkui, XU Shengjun, LI Jiping, MA Yannan, JING Zhuoqiong, GUO Zhijie, ZHENG Guo. Optimization of Fermentation Conditions of Biocontrol Bacteria HMQ20YJ04 and Evaluation of Its Effect on Botrytis cinerea[J]. Chinese Journal of Biological Control, 2023, 39(6): 1418-1433.

References

[1] Guo J, Sun K, Zhang Y, et al. SlMAPK3, a key mitogen-activated protein kinase, regulates the resistance of cherry tomato fruit to Botrytis cinerea induced by yeast cell wall and beta-glucan[J]. Postharvest Biology and Technology, 2021(171):171.
[2] 崔杰, 王翠翠, 刘永光, 等. 番茄果实灰霉病拮抗细菌菌株SG-11的鉴定及其抑菌作用[J]. 山东农业科学, 2022, 54(10): 104-109.
[3] 张庆萍, 王燕春, 王秀琴, 等. 番茄灰霉病发生特点及室内毒力测定[J]. 北方农业学报, 2018, 46(2): 69-72.
[4] 张体敏, 高霞, 田瑞新, 等. 番茄灰霉病的防治研究进展[J]. 园艺与种苗, 2022, 42(2): 28-29.
[5] 李昕灿, 张保华, 梁佳馨, 等. 山东部分地区番茄灰霉病菌对吡噻菌胺的抗性检测[J]. 中国植保导刊, 2022, 42(10): 70-72.
[6] 张林, 蒋细良, 杨晓燕, 等. 化学杀菌剂对木霉菌厚垣孢子萌发及菌丝生长的抑制作用[J]. 中国农学通报, 2014, 30(33): 150-155.
[7] 赵新贝, 王娟, 上官妮妮, 等. 番茄灰霉病生防细菌TD-7的鉴定、发酵条件优化及其防治效果[J]. 中国生物防治学报, 2019, 35(2): 226-239.
[8] 梅小飞, 王智荣, 阚建全. 荧光假单胞菌防治果蔬病害的研究进展[J]. 微生物学报, 2019, 59(11): 2069-2082.
[9] 刘锦霞, 李娜, 李晶, 等. 生防菌株SW11在番茄植株上的定殖能力及其对番茄灰霉病的防控效果[J]. 中国生物防治学报, 2015, 31(3): 353-360.
[10] 荆卓琼, 郭致杰, 徐生军, 等. 解淀粉芽胞杆菌HZ-6-3的筛选鉴定及其防治番茄灰霉病效果的评价[J]. 草业学报, 2020, 29(2): 31-41.
[11] 潘晓梅, 李昭煜, 石晓玲, 等. 番茄灰霉生防菌XF的筛选、鉴定及生防因子的初步探索[J]. 西北农业学报, 2019, 28(11): 1888-1895.
[12] 郭庆港, 刘高鸽, 陈秀叶, 等. 枯草芽胞杆菌HMB19198菌株抑菌物质的鉴定及其对番茄灰霉病的防治[J]. 植物病理学报, 2022, 52(2): 247-255.
[13] Hinarejos E, Castellano M, Rodrigo I, et al. Bacillus subtilis IAB/BS03 as a potential biological control agent[J]. European Journal of Plant Pathology, 2016, 146(3): 1-12.
[14] 方中达. 植病研究方法[M]. 北京: 中国农业出版社, 1998. 12: 191.
[15] 张菁, 连清贵, 陈婧, 等. 淡紫拟青霉QLP12对感染灰霉病后番茄植株的促生作用及抗病相关酶活性变化[J]. 植物保护学报, 2018, 45(5): 1088-1095.
[16] 张宁, 张晶晶, 李雅淑, 等. 人参黑斑病生防菌的分离鉴定及防效[J]. 中国生物防治学报, 2022, 38(5): 1308-1315.
[17] 石玉莹, 宋海慧, 苗爽, 等. 番茄灰霉病和叶霉病拮抗细菌WXCDD51的筛选鉴定及其生防促生作用[J]. 园艺学报, 2017, 44(10): 1925-1936.
[18] 齐爱勇, 魏东盛, 刘大群, 等. 番茄灰霉病拮抗细菌的筛选[J]. 中国农学通报, 2006(6): 311-313.
[19] Li Y Q, Song K, Li Y C, et al. Statistical culture-based strategies to enhance chlamydospore production by Trichoderma harzianum SH2303 in liquid fermentation[J]. Journal of Zhejiang University(Science B), 2016, 17(8): 619-627.
[20] Beneduzi A, Ambrosini A, Passaglia L M P. Plant growth-promoting rhizobacteria (PGPR): Their potential as antagonists and biocontrol agents[J]. Genetics and Molecular Biology, 2012, 35: 1044-1051.
[21] 乔俊卿, 张心宁, 梁雪杰, 等. 枯草芽胞杆菌PTS-394诱导番茄对灰霉病的系统抗性[J]. 中国生物防治学报, 2017, 33(2): 219-225.
[22] 曲远航, 郭庆港, 李社增, 等. 生防枯草芽胞杆菌HMB19198发酵培养基的筛选及优化[J]. 农药学学报, 2022, 24(3): 509-519.
[23] 杨利敏, 仝赞华, 郭立华, 等. 番茄灰霉生防菌CQ的分子鉴定及其生防效果研究[J]. 中国生物防治学报, 2015, 31(6): 956-960.
[24] 王伟, 李佳, 姜军坡, 等. 番茄灰霉病拮抗菌TB-12菌株发酵条件优化及其生防效果[J]. 北方园艺, 2017(15): 36-41.
[25] Nunes C, Usall J, Teixidó N, et al. Biological control of posthar-vest pear diseases using a Bacterium, Pantoea agglomerans CPA-2[J]. International Journal of Food Microbiology, 2001, 70(1-2): 53-61.
[26] Sun P P, Cui J C, Jia X H, et al. Isolation and characterization of Bacillus amyloliquefaciens L-1 for biocontrol of pear ring rot[J]. Horticultural Plant Journal, 2017, 3(5): 183-189.
[27] 张梦君, 黎继烈, 申爱荣, 等. 亚麻立枯病拮抗菌的筛选、生防效果及发酵条件优化[J]. 微生物学通报, 2017, 44(5): 1099-1107.
[28] 丁翠珍, 裘季燕, 刘伟成, 等. 枯草芽胞杆菌B02产生拮抗物质培养基及发酵条件优化[J]. 中国生物防治, 2008(2): 159-163.
[29] 叶云峰, 黎起秦, 袁高庆, 等. 枯草芽胞杆菌B47菌株高产抗菌物质的培养基及发酵条件优化[J]. 微生物学通报, 2011, 38(9): 1339-1346.
[30] 李术娜, 姜军坡, 朱莹, 等. 大丽轮枝菌拮抗细菌枯草芽胞杆菌12-34菌株发酵产抗菌蛋白的条件优化[J]. 植物保护, 2009, 35(3): 51-56.
[31] 何明川, 曾舒泉, 王志江, 等. 一株烟草疫霉拮抗菌MC4-2的鉴定、发酵条件优化及防效测定[J]. 微生物学通报, 2021, 48(12): 4636-4648.
[32] 张梦君, 黎继烈, 申爱荣, 等. 亚麻立枯病拮抗菌的筛选、生防效果及发酵条件优化[J]. 微生物学通报, 2017, 44(5): 1099-1107.
[33] 高越, 马贵龙, 臧超群, 等. 葡萄炭疽病生防菌株GS8发酵培养基及发酵条件的优化[J]. 吉林农业大学学报, 2015, 37(4): 411-416.
[34] 陶树兴, 丛寅, 徐姗娜, 等. 枯草芽胞杆菌SNUB16的培养条件及抗真菌物质[J]. 陕西师范大学学报(自然科学版), 2012, 40(4): 56-60.
[35] 赵志祥, 严婉荣, 陈圆, 等. 一株生姜青枯病拮抗菌的筛选、鉴定及发酵条件优化[J]. 南方农业学报, 2015, 46(3): 421-427.
[36] 梁雪杰, 刘邮洲, 张荣胜, 等. 生防菌PTS-394发酵培养基及培养条件的优化[J]. 扬州大学学报(农业与生命科学版), 2013, 34(3): 75-82.
[37] Lin X, Ye J, Mu C, et al. Research on the fermentation conditions of Bacillus subtilis B-332[J]. Agricultural Science & Technology, 2013, 14(1): 81.