This study investigated the antifungal activity and possible mode of action of Bacillus pumilus HN-10 antifungal peptide P-1 against Trichothecium roseum.The results showed that the antifungal peptide P-1 at a concent...This study investigated the antifungal activity and possible mode of action of Bacillus pumilus HN-10 antifungal peptide P-1 against Trichothecium roseum.The results showed that the antifungal peptide P-1 at a concentration of 1.0μg mL^(-1)had strong antifungal activity against T.roseum.P-1 inhibited the tricarboxylic acid cycle(TCA)pathway and the transporter pathway of NADH to coenzyme Q on the electron transport chain.P-1 significantly reduced succinate dehydrogenase(SDH),malate dehydrogenase(MDH),ATPase,mitochondrial complex enzymes I,II and IV enzyme activities on the electron transport chain,and 5'-triphosphate(ATP),5'-diphosphate(ADP),5'-monophosphate(AMP)content,and energy charge(EC);significantly increased 6-phosphofructokinase(PFK)enzyme activity.The release of Ca^(2+)(OD_(680))from the inner mitochondrial membrane and the openness of the mitochondrial permeability transition pore(MPTP)were analysed,and microscopy was performed following staining of mitochondria with JC-1.The results indicated that P-1 significantly increased the release of Ca^(2+) and the openness of MPTP,decreased the mitochondrial membrane potential,and produced green fluorescence;transcriptomics data analysis showed that there were 39 differentially expressed genes(DEGs)related to energy metabolism enzymes.The results verified by qRT-PCR were basically consistent with the transcriptome sequencing results.Thus,P-1 achieved its inhibitory effect mainly by regulating genes related to energy metabolism.展开更多
基金financially supported by National Key R&D Program of China(2018YFD0400205).
文摘This study investigated the antifungal activity and possible mode of action of Bacillus pumilus HN-10 antifungal peptide P-1 against Trichothecium roseum.The results showed that the antifungal peptide P-1 at a concentration of 1.0μg mL^(-1)had strong antifungal activity against T.roseum.P-1 inhibited the tricarboxylic acid cycle(TCA)pathway and the transporter pathway of NADH to coenzyme Q on the electron transport chain.P-1 significantly reduced succinate dehydrogenase(SDH),malate dehydrogenase(MDH),ATPase,mitochondrial complex enzymes I,II and IV enzyme activities on the electron transport chain,and 5'-triphosphate(ATP),5'-diphosphate(ADP),5'-monophosphate(AMP)content,and energy charge(EC);significantly increased 6-phosphofructokinase(PFK)enzyme activity.The release of Ca^(2+)(OD_(680))from the inner mitochondrial membrane and the openness of the mitochondrial permeability transition pore(MPTP)were analysed,and microscopy was performed following staining of mitochondria with JC-1.The results indicated that P-1 significantly increased the release of Ca^(2+) and the openness of MPTP,decreased the mitochondrial membrane potential,and produced green fluorescence;transcriptomics data analysis showed that there were 39 differentially expressed genes(DEGs)related to energy metabolism enzymes.The results verified by qRT-PCR were basically consistent with the transcriptome sequencing results.Thus,P-1 achieved its inhibitory effect mainly by regulating genes related to energy metabolism.
