Functional Research On Genes TaFet1,TaGl1 And TaLaC1 Of Trichoderma Atroviride

Trichoderma atroviride parasitizes a variety of plant pathogenic fungi and is widely used for biocontrol of plant fungal diseases.It can also cause postharvest rot of citrus.Previous,genes specifically expressed during the infection of citrus in T.atroviride were identified through RNA-seq technology.In this study,the function of copper-oxidase genes（TaFET1 and TaLaC1）and a GH64 family gene（TaGl1）were analyzed through PEG mediated protoplast transformation.The results are as follows:Gene TaFet1 is 1845 nt length and encodes a 614-amino-acid ferroxidase.The functional analysis showed that the disruption of TaFet1 did not affect the vegetative growth,parasitism against mycelia of Sclerotinia sclerotiorum.However,its sporulation,laccase activity and pathogenicity to citrus fruit decreased by 100%,81.5%,and 39.6%,respectively.In addition,TaFet1 knockout also influenced cell integrity and increased the sensitivity to H₂O₂.The overexpression of TaFet1 increased the amount of conidia by 32%and the sensitivity to cell wall inhibitors,while it had no significant influence on the mycelial growth rate,colony morphology,pathogenicity to citrus fruit,and parasitism against mycelia of S.sclerotiorum.Thus,these results indicate TaFet1is involved in the infection on citrus fruit.Gene TaLaC1 has a total length of 1921 nt and encodes a 590-amino-acid laccase.The deletion of TaLaC1 did not affect the vegetative growth,sporulation,cell integrity,pathogenicity to citrus fruits,laccase activity,or parasitic ability to sclerotia and mycelia of S.sclerotiorum.Gene TaGl1 has a total length of 1378 nt and encodes a glycoside hydrolase of450-amino-acid with a signal peptide of 16 amino acids.Deletion of TaGl1 did not affect the vegetative growth,sporulation,cell integrity,pathogenicity to citrus fruits,or the parasitic ability to sclerotia and mycelia of S.sclerotiorum.In this study,the function of copper-oxidase genes（TaFet1 and TaLaC1）and a GH64 family gene（TaGl1）were studied.The results will help understand the mechanism of T.atroviride to interact with different hosts.