| Botrytis cinerea is a non-host-specific phytopathogenic fungus,which can cause gray mold on most cash crops such as fruits,vegetables and ornamental flowers,thus causing significant economic losses.The clarification of the mechanism of conidial production and pathogenesis is of great importance to the development of new and efficient agents for the prevention and control of gray mold.In our preliminary study,we discovered the Bcb1 gene,which encodes a protein of 541 aa with an AMP-binding domain,by a pair of qPCR primers for B.cinerea with good stability and specificity.Bioinformatic analysis of the Bcb1protein suggested that it is a membrane protein belonging to the Acyl coenzyme A synthetase family.This class of proteins regulates the conidial production and pathogenicity in Magnaporthe oryzae and Beauveria bassiana.Above all,we conducted a preliminary study on the function of the Bcb1 gene using a reverse genetics approach,and the main results are as follows.1,Using a overlap PCR-based split marker strategy and PEG/CaCl2-mediated transformation of protoplasts,the Bcb1 gene was knocked out by replacement with a neomycin-resistant Neo gene,and two Bcb1 gene deletion mutants were obtained by PCR,Southern blot and RT-PCR validation.2,Compared with the wild-type strain,the deletion of Bcb1 gene resulted in a dramatic slowdown in the growth rate of the strain,reduced aerial mycelium,impaired conidial production and melanin formation ability,altered the sensitivity of the strain to oxidative stress,osmotic stress caused by KCl,impaired cell wall and cell membrane integrity and the ability of infection cushion formation of the strain,and reduced pathogenicity.3,This gene positively regulates melanin formation-related genes Bcpks12 and Bcpks13,polygalacturonase-encoding gene Bcpg2,phytotoxin botrydial synthesis-related gene BcBOT2,cutinase-encoding gene cutA,and the genes Bc1G＿12707.1and BcPDR1,which affect processes related to the growth,development and pathogenicity of B.cinerea. |
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