Functions of CfPOFUT1 in Colletotrichum fructicola

Abstract:

[Objective] Protein O-fucosyltransferase 1(POFUT1),a key enzyme catalyzing protein O-fucosylation,has been verified to regulate a series of physiological and pathological processes in animals and humans.However,POFUT1 gene in Colletotrichum fructicola or even fungi has not been reported.This study aims to clone the CfPOFUT1 gene from C.fructicola and analyze its biological functions.[Methods] RT-PCR was employed to amplify the CfPOFUT1 gene and then the bioinformation of the gene was analyzed.We constructed the CfPOFUT1 silencing and overexpression vectors,which were then transformed into the protoplasts with the PEG-mediated method to yield the CfPOFUT1-silenced and-overexpressing mutants.The biological phenotypes of the wild-type LY5-1,and the CfPOFUT1-silenced and-overexpressing mutants were determined,such as the mycelia growth,conidial production,conidial germination,appressorium formation,stress response,pathogenicity,and fungicide sensitivity.[Results]CfPOFUT1-overexpressing mutant showed higher conidial production,stronger pathogenicity,lower sensitivity to azoxystrobin,but higher sensitivity to carbendazim and prochloraz than the wild-type LY5-1.CfPOFUT1-silenced mutant demonstrated lower conidial production,intact cell wall,higher sensitivity to endoplasmic reticulum (ER) stress and azoxystrobin,lower pathogenicity,and lower sensitivity to carbendazim and prochloraz.[Conclusion] CfPOFUT1 is involved in conidial production,cell wall integrity,ER stress,and fungicide sensitivity of C.fructicola,affecting the pathogenicity.