Optimization of Fermentation Culture Medium and Flask Fermentation Conditions for Bacillus thuringiensis Strain JQD117 with High Toxicity against Bradysia odoriphaga

doi:10.16409/j.cnki.2095-039x.2021.07.013

Abstract

Abstract: Aiming to increase living spore production of a Bacillus thuringiensis (Bt) strain JQD117, we employed response surface analysis methods to optimize the culture medium and the flask fermentation conditions. Based on preliminary results from single factor experiments, we used response surface analysis to model and optimize the components of culture medium and parameters of flask fermentation conditions. The best ratios of each single factors for the strain Bt JQD117 were 2.00% cotton seed cake meal, 1.00% soybean meal, 1.50% yeast extract, 2.00% soluble starch, 0.30% CaCO₃, 0.04% MnSO₄, 0.18% MgSO₄, and 0.04% K₂HPO₄. For fermentation conditions, the optimum parameters were 3% inoculation amount, 60 mL liquid volume in flask, 150 r/min at 30 ℃, and pH 7.5. Theoretically, the living spore can reach 6.75×10⁹ spore/mL under this optimized condition. The verification experiment showed that the living spore yield was 5.97×10⁹ spore/mL. Our results have placed a critical foundation and theoretical basis for large-scale application of strain JQD117 for green control of Bradysia odoriphaga and the safe production of vegetables such as leeks.

Key words: Bradysia odoriphaga, Bacillus thuringiensis, fermentation, single-factor experiment, response surface analysis

CLC Number:

S476.1

SONG Jian, Zhang Haijian, FENG Shuo, CHENG Jiaxu, Liu Jianhu, Guo Weibing, CAO Weiping. Optimization of Fermentation Culture Medium and Flask Fermentation Conditions for Bacillus thuringiensis Strain JQD117 with High Toxicity against Bradysia odoriphaga[J]. Chinese Journal of Biological Control, 2022, 38(2): 333-341.

References

[1] 王哲, 钟涛, 刘培斌, 等. 韭菜迟眼蕈蚊发生规律及防治方法研究进展[J]. 环境昆虫学报, 2017, 39(6): 1397-1406.
[2] Zhang P, Zhao Y H, Wang Q H, et al. Lethal and sublethal effects of the chitin synthesis inhibitor chlorfluazuron on Bradysia odoriphaga Yang and Zhang (Diptera: Sciaridae)[J]. Pesticide Biochemistry and Physiology, 2017, 136: 80-88.
[3] 党志红, 董建臻, 高占林, 等. 不同种植方式下韭菜迟眼蕈蚊发生为害规律的研究[J]. 河北农业大学学报, 2001, 24(4): 65-68.
[4] Li W X, Yang Y T, Xie W, et al. Effects of temperature on the age-stage, two-sex life table of Bradysia odoriphaga (Diptera: Sciaridae)[J]. Journal of Economic Entomology, 2015, 108: 126-134.
[5] Yabuki Y, Mukaida Y, Saito Y, et al. Characterisation of volatile sulphur-containing compounds generated in crushed leaves of Chinese chive (Allium tuberosum Rottler)[J]. Food Chemistry, 2010, 120: 343-348.
[6] Hare J D. Ecology and management of the Colorado potato beetle[J]. Annual Review of Entomology, 1990, 35: 81-100.
[7] Zhang X B, Candas M, Criko N B, et al. A mechanism of cell death involving an adenylyl cyclase/PKA signaling pathway is induced by the Cry1Ab toxin of Bacillus thuringiensis[J]. Proceedings of the National Academy of Sciences of the United States of America, 2006, 103(26): 9897-9902.
[8] Sanchis V. From microbial sprays to insect-resistant transgenic plants: history of the biospesticide. A review[J]. Agronomy for Sustainable Development, 2011, 30(1): 217-231.
[9] Favret M E, Yousten A A. Thuricin: the bacteriocin produced by Bacillus thuringiensis[J]. Journal of Invertebrate Pathology, 1989, 53(2): 206-216.
[10] Cherif A, Rezgui W, Raddadi N, et al. Characterization and partial purification of entomocin 110, a newly identified bacteriocin from Bacillus thuringiensis subsp. Entomocidus HD110[J]. Microbiological Research, 2008, 163: 684-692.
[11] Nester E W, Thomashow L S, Metthew M, et al. 100 Years of Bacillus thuringiensis: A critical scientific assessment[M]//American Academy of Microbiology, 2002.
[12] Fred S B, Bruce G H, Roy L F. Safety and advantages of Bacillus thuringiensis-protected plants to control insect pests-ScienceDirect[J]. Regulatory Toxicology and Pharmacology, 2000, 32(2): 156-173.
[13] 任羽, 郭文超. 苏云金芽胞杆菌在马铃薯甲虫防治上的研究进展[J]. 植物保护学报, 2017, 44(5): 705-712.
[14] 喻子牛. 苏云金杆菌[M]. 北京:科学出版社, 1990, 285-423.
[15] 陈宇熹, 叶舒婷, 蔡美凤, 等. 高效杀蚊苏云金芽孢杆菌BRC-LLP29的发酵优化[J]. 生物数学学报, 2012, 27(1): 175-185.
[16] 成雪飞, 宋志强, 王剑, 等. 响应面法优化苏云金芽胞杆菌YC-10发酵培养基[J]. 湖南师范大学学报(自然科学版), 2016, 39(4): 23-28.
[17] 张路路, 朱朝华, 郭刚. 苏云金芽孢杆菌A322菌株发酵培养基和发酵条件的优化[J]. 热带生物学报, 2014, 5(3): 253-259.
[18] 宋健, 曹伟平, 杜立新. 对韭蛆高毒力Bt菌株与常用化学杀虫剂相容性研究[J]. 中国生物防治学报, 2017, 33(6): 739-746.
[19] 龚军辉, 王晶. 稀释涂布平板法计数活菌的方法简介[J]. 生物学教学, 2018, 43(2): 70-71.
[20] 兰周. 多杀菌素生物合成基因簇的克隆及异源表达载体的构建[D]. 福州:福建农林大学, 2012.
[21] Sumant P, Qasim K B, Rani G. Optimization of alkaline protease production from Bacillus sp. by response surface methodology[J]. Current Microbiology, 2002, 44: 286-290.
[22] 周虓, 郑毅, 叶海梅, 等. 响应面分析法优化耐高温蛋白酶发酵培养基[J]. 生物数学学报, 2007, 22(1): 113-118.
[23] 郑毅, 周堍, 黄勤清, 等. 产耐温蛋白酶苏云金芽孢杆菌FS140 液体发酵条件优化[J]. 应用与环境生物学报, 2007, 13(5): 708-712.
[24] 杨梅, 张峰, 苏新华. 苏云金芽孢杆菌LLB19 发酵培养基的优化[J]. 福建师范大学学报, 2009, 25(2): 75-81.
[25] 张群林, 张志国, 余洁, 等. 响应面分析法优化苏云金芽孢杆菌BRC-ZQL3 菌株发酵培养基[J]. 福建师范大学学报, 2011, 27(5): 85-90.
[26] 李姝江, 王淋敏, 谯天敏, 等. 利用响应面法优化贝莱斯芽孢杆菌ZJ20发酵参数[J]. 西北农林科技大学学报(自然科学版), 2019, 47(2): 88-96.