ARTP诱变灰葡萄孢霉筛选高产脱落酸菌株

Selection of High-Yield Abscisic Acid Strains Through Atmospheric Room Temperature Plasma Mutagenesis

目的:利用常压室温等离子体(Atmospheric Room Temperature Plasma,ARTP)技术对生产脱落菌的潜力宿主灰葡萄孢霉进行诱变,以获得遗传稳定性好的高产脱落酸的突变菌种。方法:利用酶解法去除灰葡萄孢霉细胞壁,从而获得灰葡萄孢霉原生质体。接着将灰葡萄孢霉原生质体在输出功率120 W,工作气流10 L/min,照射距离2 mm,照射时间120 s的条件下进行ARTP诱变处理。检测突变菌株的脱落酸产量,并对最佳突变菌株进行发酵优化。结果:与野生型相比,突变菌株的脱落酸生产能力有了不同程度的提高,其中突变菌株A4和A7的脱落酸生产能力显著提高,分别提高了26.3%和30.5%。进一步通过单因素实验,确定突变菌株A7的最佳发酵碳源和氮源成分为葡萄糖和黄豆粉,最佳接种量为8%。结论:采用ARTP诱变技术能够有效获得高产脱落酸的突变菌株,为脱落酸的工业化生产提供了有利支撑。

Objective:To utilize atmospheric room temperature plasma(ARTP)technology to induce mutations in Botrytis cinerea,a potential host for abscisic acid production,aiming to obtain genetically stable mutant strains with high abscisic acid titer.Methods:A mixed enzyme solution is used to remove the cell wall of B.cinerea to obtain protoplasts.The protoplasts are then subjected to ARTP mutagenesis treatment under conditions of 120 W output power,10 L/min working gas flow,2 mm irradiation distance,and 120 s irradiation time.Finally,the abscisic acid yield of the mutant strains is measured,and fermentation optimization is performed on the best mutant strain.Results:Compared to the wild-type strain,the mutant strains show varying degrees of improvement in abscisic acid production.Notably,mutant strains A4 and A7 exhibit significant increases in abscisic acid production,with enhancements of 26.3%and 30.5%,respectively.Further single-factor experiments determined that the optimal fermentation carbon and nitrogen sources for mutant strain A7 are glucose and soybean meal,with an optimal inoculation amount of 8%.Conclusion:The mutant strains with high abscisic acid production can be effectively obtained by using ARTP mutagenesis technology,which provides favorable support for the industrial production of abscisic acid.