Beauveria bassiana is a promising biocontrol agent due to its entomopathogenic activities and residue-free characteristics. However, its susceptibility to abiotic stresses and naturally low virulence limit the effective application of this fungus. To effectively obtain fungal strains with high biocontrol potential, fluorescence-activated cell sorting (FACS) was used to screen mutant libraries generated by atmospheric and room temperature plasma (ARTP). Among about 8000 mutants obtained by ARTP mutagenesis, six candidate mutants were selected according to the forward scatter (FSC) signal readings of FACS. B6, with a 37.4% higher FSC reading than wild-type (WT), showed a 32.6% increase in virulence. It also presented a 13.5% decrease in median germinating time (GT50) and a 12.1% increase in blastospore production. Comparative analysis between insect transcriptional responses to B6 and WT infection showed that the immune response coupled with protein digestion and absorption progress was highly activated in B6-infected Galleria mellonella larvae, while fatty acid synthesis was suppressed after 3 days of infection. Our results confirmed the feasibility of sorting B. bassiana with high biocontrol potential via the combination of ARTP and FACS and facilitated the understanding of insect-pathogen interactions, highlighting a new strategy for modifying entomopathogenic fungi to improve the efficiency of biological control.

中文翻译：

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基于ARTP诱变和高通量FACS筛选具有高生防潜力的白僵菌

白僵菌因其昆虫病原活性和无残留特性而成为一种很有前途的生物防治剂。然而，它对非生物胁迫的敏感性和天然的低毒力限制了这种真菌的有效应用。为了有效地获得具有高生防潜力的真菌菌株，荧光激活细胞分选（FACS）被用于筛选由常温和室温等离子体（ARTP）产生的突变文库。在通过ARTP诱变获得的约8000个突变体中，根据FACS的前向散射（FSC）信号读数选择了6个候选突变体。B6 的 FSC 读数比野生型 (WT) 高 37.4%，毒力增加了 32.6%。它还表现出中位发芽时间 (GT50) 减少 13.5% 和芽生孢子产量增加 12.1%。昆虫对 B6 和 WT 感染的转录反应的比较分析表明，在 B6 感染的大蜡螟幼虫中，结合蛋白质消化和吸收过程的免疫反应高度激活，而感染 3 天后脂肪酸合成受到抑制。我们的结果证实了通过 ARTP 和 FACS 的组合对具有高生物防治潜力的 B. bassiana 进行分类的可行性，并促进了对昆虫-病原体相互作用的理解，突出了一种修改昆虫病原真菌以提高生物防治效率的新策略。而脂肪酸合成在感染 3 天后被抑制。我们的结果证实了通过 ARTP 和 FACS 的组合对具有高生物防治潜力的 B. bassiana 进行分类的可行性，并促进了对昆虫-病原体相互作用的理解，突出了一种修改昆虫病原真菌以提高生物防治效率的新策略。而脂肪酸合成在感染 3 天后被抑制。我们的结果证实了通过 ARTP 和 FACS 的组合对具有高生物防治潜力的 B. bassiana 进行分类的可行性，并促进了对昆虫-病原体相互作用的理解，突出了一种修改昆虫病原真菌以提高生物防治效率的新策略。