常压室温等离子体(ARTP)诱变快速选育对亚洲玉米螟高毒的苏云金芽孢杆菌突变株
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摘要
采用常压室温等离子体(Atmospheric and room temperature plasma,ARTP)诱变苏云金芽孢杆菌Bt-NBIC-380菌株,选育对亚洲玉米螟高毒力突变株。经过4轮的ARTP诱变、镜检及摇瓶发酵筛选得到的突变株Bt-NBIC-380-18-407,其摇瓶发酵液对玉米螟二龄幼虫效价达到6 835 IU/μL,杀虫晶体蛋白含量6.3 mg/mL。该突变株的效价较初始菌株提高了25.7%,杀虫晶体蛋白含量提高了18.9%。经过5次传代,证实该突变株具有良好的遗传稳定性。
A highly toxic Bacillus thuringiensis strain NBIC-380 against Asian corn borer was treated by atmospheric and room temperature plasma for improving the toxicity of the resulting mutants. Through 4 rounds of ARTP mutation and screening by microscopic checking,shake-flask fermentation and bioassay,one mutant designated as Bt-NBIC-380-18-407 with significant improvement of production of insecticidal crystal protein and potency-was obtained. The potency of the fermentation broth of the mutant against Asian corn borer larvae was increased 25.7% to 6 835 IU/μl,while the concentration of insecticidal crystal protein in fermentation broth was increased 18.9% to 6.3 mg/mL. The mutant showed rather good genetic stability after 5 times of consecutive transferring and cultivation.
A highly toxic Bacillus thuringiensis strain NBIC-380 against Asian corn borer was treated by atmospheric and room temperature plasma for improving the toxicity of the resulting mutants. Through 4 rounds of ARTP mutation and screening by microscopic checking,shake-flask fermentation and bioassay,one mutant designated as Bt-NBIC-380-18-407 with significant improvement of production of insecticidal crystal protein and potency-was obtained. The potency of the fermentation broth of the mutant against Asian corn borer larvae was increased 25.7% to 6 835 IU/μl,while the concentration of insecticidal crystal protein in fermentation broth was increased 18.9% to 6.3 mg/mL. The mutant showed rather good genetic stability after 5 times of consecutive transferring and cultivation.
引文
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[5] GB/T 19567.1-2004苏云金芽胞杆菌原粉[S].
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[2]胡金宏,杨树林.浅谈玉米病虫害的综合防治技术要点[J].农业与技术,2016,36(12):114-115.
[3]田淑翠,牛延宁,常忠义,等.常压室温等离子体诱变茂源链霉菌菌株[J].中国生物工程杂志,2016,36(9):47-53.
[4]王耀耀,朱研研,付美红,等.那他霉素产生菌的ARTP的诱变育种及发酵工艺优化[J].中国医药工业杂志,2016,47(8):999-1004.
[5] GB/T 19567.1-2004苏云金芽胞杆菌原粉[S].
[6]周荣华,廖先清,刘芳,等.玉米螟高毒力生防菌株筛选与发酵工艺研究[J].湖北农业科学,2015,54(24):6248-6251.
[7]贾春生.利用SPSS软件计算杀虫剂的LC50[J].昆虫知识,2006,43(3):414-417.
[8]马娜娜,马韵升,张心青,等.一株产胶霉菌素菌株的ARTP诱变及筛选[J].中国农学通报,2017,33(29):36-41.
[9]杜丹清,王刚,庹有朋,等.γ-聚谷氨酸生产菌种的ARTP诱变及其发酵条件优化[J].食品与发酵科技,2017,53(6):5-10.