SEM and qRT-PCR revealed quercetin inhibits morphogenesis of Aspergillus flavus conidia via modulating calcineurin-Crz1 signalling pathway

Abstract

Aspergillus flavus exploits diverse mechanisms to survive during exposure to antifungal agents including morphogenesis. Germination of dormant conidia involves cascades of reactions integrated into the signalling pathway. This study documents the effect of phytochemical-quercetin on A. flavus during germination of conidia using scanning electron microscopy (SEM). Significant inhibition of conidial swelling of A. flavus in comparison to control was observed at 4 and 7 h Quantitative real-time PCR for genes from calcium signalling pathway and heat-shock proteins family showed up-regulation of heat shock (Hsp70 and Hsp90) and calcium signalling pathway genes (calcium-transporting ATPase and calmodulin) in response to quercetin at initial 4 h in comparison to control sample whereas up-regulation of Hsp70, calcineurin and transcription factor Crz1, were observed in both the treated samples. Gene encoding for calcium-kinase, cAMP, Rho-gdp, Plc and Pkc showed a constitutively higher level of expression in quercetin-treated sample in comparison to control at both time points. These data showed a clear response from genes encoding calcineurin-Crz1 signalling pathways and may find its application in the screening of antifungal agents. Abbreviations Hsp: Hear shock protein; MIC: Minimum Inhibitory Concentration; SEM: Scanning Electron Microscopy; qRT-PCR: Quantitative Real-Time Polymerase Chain Reaction

Keywords

• aspergillus flavus
• scanning electron microscopy
• qrt-pcr
• calcium signalling pathway
• quercetin