中国生物防治学报2017，Vol.33Issue(4)：537-544,8.DOI:10.16409/j.cnki.2095-039x.2017.04.015
利用双亲灭活原生质体融合技术选育高效苏云金芽胞杆菌和枯草芽胞杆菌融合菌株
Selection of Insecticidal and Fungicidal Engineering Strain by Protoplasts Fusion of Inactivated Parental Strain Bacillus thuringiensis and Bacillus subtilis
摘要
Abstract
Fermentation broth of Bacillus thuringiensis B7 caused 85.06％ mortality ofPlutella xylostella 2nd instar larvae at 96 h after treatment.Bacillus subtilis TL2 showed antagonistic effects on pathogenic fungi Fusarium oxysporum and Phytophthora capsici,which reduced more than 60％ of mycelial growth of these fungi at 5 d after confronting incubation.The best condition for protoplast obtaining,complete inactivation,and protoplast fusion of these two bacterial strains were determined.The best protoplast formation rate and regeneration rate were 85.00％ and 29.41％,respectively,for strain B7 under 1.0 mg/L of lysozyme for 70 min.The best protoplast formation rate and regeneration rate were 80.00％ and 25.00％,respectively,for strain TL2 under 1.25 mg/L of lysozyme for 50 min.The best protoplasts inactivation condition for strain TL2 was water bath at 60 ℃ for 35 min,and for strain B7,ultraviolet radiation for 25min.The best protoplast fusion condition was water bath at 30 ℃ for 20 min.Then the mixture was spread on regeneration medium plates.Seventeen fusants were obtained after 10 times of subcultures,and the fusant TLB 15 showed both high insecticidal and fungicidal activity.Fermentation broth of TLB 15 resulted in 93.44％ mortality of P.xylostella 2nd instar larvae at 96 h after treatment and inhibited 60.00％ and 62.79％ of mycelial growth of F.oxysporum and P capsici,respectively (5 d of confrontation cultivation).关键词
原生质体融合/枯草芽胞杆菌/苏云金芽胞杆菌Key words
protoplast fusion/Bacillus subtilis/Bacillus thuringiensis分类
农业科技引用本文复制引用
赵晓燕,周方园,周红姿,吴晓青,张广志,张新建,李天元,谢雪迎..利用双亲灭活原生质体融合技术选育高效苏云金芽胞杆菌和枯草芽胞杆菌融合菌株[J].中国生物防治学报,2017,33(4):537-544,8.基金项目
国家自然科学基金(31572044) （31572044）
山东省优秀中青年科学家科研奖励基金(BS2015 SW029):山东省攻关项目(2010GSF10206) （BS2015 SW029）
