基于响应面法的双向伯克霍尔德氏菌BK发酵培养基优化  

Optimization of Burkholderia ambifaria(BK)fermentation medium based on response surface methodology

作　　者：戍柯婷 韩小红 周柏威 齐文忠 齐欢 吴松青[1,3] SHU Keting;HAN Xiaohong;ZHOU Bowei;QI Wenzhong;QI Huan;WU Songqing(College of Forestry,Fujian Agriculture and Forestry University,Fuzhou 350002,China;College of Plant Protection,Fujian Agriculture and Forestry University,Fuzhou 350002,China;Key Laboratory of Ecological Forestry Pest Management,Fujian Province University,Fujian Agriculture and Forestry University,Fuzhou 350002,China;Jukou Forestry Station of Yanping District,Nanping City,Nanping 353000,China;Fujian Forestry Pest Control and Quarantine Bureau,Fuzhou 350003,China)

机构地区：[1]福建农林大学林学院,福建福州350002 [2]福建农林大学植物保护学院,福建福州350002 [3]福建农林大学生态公益林重大有害生物防控福建省高校重点实验室,福建福州350002 [4]南平市延平区巨口林业站,福建南平353000 [5]福建省林业有害生物防治检疫局,福建福州350003

出　　处：《中国森林病虫》2024年第5期40-47,共8页Forest Pest and Disease

基　　金：国家重点研发计划项目“松材线虫传播媒介高效防控产品技术研发与示范”(2021YFD1400905);国家自然科学基金面上项目“基于功能域替换的松墨天牛靶向毒素改造及其作用机制”(KAA21022A);福建省林业科技项目“松墨天牛高效生物防治新技术应用与示范”(2024FKJ11);科技支撑计划一般项目“贵州省松材线虫病侵染初期的瞬时迁移规律及其综合防治技术研究”(黔科合支撑[2024]一般083)。

摘　　要：为探究马尾松Pinus massoniana的伴生细菌双向伯克霍尔德氏菌BK菌株的生防潜力,优化其发酵培养基。通过单因素试验筛选得到的最佳碳源为蔗糖和麦芽糖,最佳氮源为酵母粉和胰蛋白胨,最佳无机盐为硫酸镁和柠檬酸三钠;利用Plackett-Burman试验确定3个主要影响因素分别为蔗糖、麦芽糖和装液量;通过最陡爬坡试验结合Box-Behnken设计试验及响应面分析法,以发酵液OD_(600)为响应值,优化培养基配比,获得培养基最佳配方为蔗糖8.0 g/L,麦芽糖12.5 g/L,酵母粉11.3 g/L,硫酸镁0.6 g/L,氯化钠7 g/L;最佳发酵条件为在30℃、150 r/min条件下发酵、装液量25 mL/250 mL、接菌量1%,pH7.0~7.2;根据最佳配比验证,优化后培养基的OD_(600)为1.25,较初始培养基(OD_(600)=0.62)提高1.02倍,与模型预测值相符。研究结果优化了双向伯克霍尔德氏菌BK菌株生产发酵条件,可为后续该菌的实践应用提供理论基础。To explore the biological control potential of Burkholderia ambifaria(BK),the associated bacterium of Pinus massoniana,the medium was optimized.The best carbon sources were sucrose and maltose,the best nitrogen sources were yeast extract and tryptone,and the best inorganic salts were magnesium sulfate(MgSO_(4))and trisodium citrate(C_(6)H_(9)Na_(3)O_(9))by single‑factor test screeneing.The Plackett-Burman test determined the three main influencing factors were sucrose,maltose and liquid volume.The steepest ascent test combining with the Box-Behnken design and response surface analysis method were used,and using the optical density at 600 nm(OD_(600))of the fermentation broth as the response variable,the composition of the medium was optimized around optimal concentrations.The optimal formula of the medium were sucrose 8.0 g/L,maltose 12.5 g/L,yeast extract 11.3 g/L,MgSO_(4)0.6 g/L,NaCl 7 g/L.The optimal fermentation conditions were at 30℃and 150 r/min,the liquid volume of 25 mL/250 mL,the inoculation amount of 1%,a pH range of 7.0 to 7.2.Verification of the optimal ratio indiacated,that the OD_(600)of the optimized medium was 1.25,which was 1.02 times higher than that of the initial medium(OD_(600)=0.62),which was consistent with the predicted value of the model.The results of this study optimized the fermentation conditions for the production of Burkholderia ambifaria(BK),which could provide a theoretical basis for the practical application of the strain.

关 键 词：双向伯克霍尔德氏菌 单因素试验 PLACKETT-BURMAN设计 OD_(600) 

分 类 号：S763.306.4[农业科学—森林保护学]