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通过敲除球孢白僵菌(Beauveria bassiana)菌株bm01的PacC基因,研究了PacC基因敲除对球孢白僵菌毒力和分生孢子萌发速率的影响。利用农杆菌(Agrobacterium tumefaciens)介导的转化方法转化Bb_bm01,通过同源重组敲除Bb_bm01的PacC基因,筛选ΔPacC基因敲除菌株,获得了遗传稳定的ΔPacC基因敲除菌株。通过毒力测定,对比了野生菌株和ΔPacC基因敲除菌株对家蚕和大蜡螟毒力的差异,发现ΔPacC基因敲除菌株对家蚕和大蜡螟毒力都减弱,差异具有统计学意义(p<0.01),表明在Bb_bm01菌株中的PacC基因与它的致病性相关;另外,比较ΔPacC基因敲除菌株与野生型菌株Bb_bm01在SDB液体培养基中的孢子萌发率,发现ΔPacC基因敲除菌株孢子萌发明显减慢,表明PacC基因参与调控孢子萌发。
The PacC gene knockout of Beauveria bassiana strain bm01 was used to study the effect of PacC knockout on the virulence of Beauveria bassiana and the conidial germination rate. The Agrobacterium tumefaciens-mediated transformation method was used to transform Bb_bm01. The PacC gene of Bb_bm01 was knocked out by homologous recombination and the ΔPacC gene knockout strain was screened. The genetically stable ΔPacC knockout strain was obtained. The virulence of wild strains and ΔPacC knockout strains was compared to the virulence of silkworm (Bombyx mori) and Mycelium lanceolata by virulence assay. It was found that the virulence of ΔPacC knockout strains was weakened to Bombyx mori and Bombyx mori, with statistical significance (p <0.01), indicating that the PacC gene in Bb_bm01 strain is related to its pathogenicity. In addition, comparing the spore germination rate of ΔPacC knockout strain and wild-type strain Bb_bm01 in SDB liquid medium, it was found that ΔPacC knockout strain Spore germination was significantly slowed down, indicating PacC gene involved in the regulation of spore germination.