摘要
氨肽酶是一类可以选择性剪切氨基酸残基的蛋白酶,液泡氨肽酶Y能够调控碱性氨基酸残基的剪切以及液泡的活性.本研究通过同源性比对获得酵母液泡氨肽酶Y在稻瘟病菌中的同源基因PoAPE3,利用基因敲除法获得PoAPE3基因敲除突变体(ΔPoAPE3),并构建回补体菌株进行生物学功能分析.表型测定结果表明,PoAPE3基因敲除突变体在产孢量、孢子萌发率、致病性、侵染过程等方面与野生型均无明显差异.突变体在完全培养基(CM)和稻秆培养基(SDC)上生长速率下降,且在应对胁迫剂H2O2时表现为更耐受,应对刚果红时表现为更敏感.以上结果表明PoAPE3参与调控稻瘟病菌的营养生长和对外界环境胁迫的应答过程.
Abstract
Aminopeptidases belong to a class of proteases capable of selectively cleaving amino acid residues.Acting as an aminopeptidase,vacuolar aminopeptidase Y regulates the cleavage of basic amino acid residues and influences vacuolar activity.In this study,the homologous gene PoAPE3,highly consistent with vacuolar aminopeptidase Y of Saccharomyces cerevisiae,was obtained from Pyricularia oryzae.Through gene knockout techniques,the ΔPoAPE3 was obtained,and a complementary strain was constructed for functional analysis.Phenotypic determination results showed that the ΔPoAPE3 exhibited no significant differences from the wild-type strain in terms of spore production,spore germination,pathogenicity,and infection process.However,ΔPoAPE3 displayed slower growth on CM and SDC media,increased tolerance to H2O2,and heightened sensitivity to Congo Red.These results indicate that PoAPE3 plays a role in vegetative growth and the response to external environmental stressors.
维普
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