Function Analysis Of BcGP Gene In Botrytis Cinerea

The ginseng(P.ginseng C.A.Meyer)is one of the most important traditional Chinese herbal medicines.It has a very high medicinal and economic value.The ginseng is a perennial medicinal plant of the Araliaceae Panax.Northeast China is an important ginseng producing area,and ginseng grey mold is one of the main diseases of ginseng industry which can destroy the leaves,petioles,flowers and fruits.The disease could seriously infect the stems and roots of ginseng.Ginseng grey mold has become one of the main hazards to the sustainable health of ginseng industry.BcGP gene may encode the transglutaminase(GATase)in B.cinerea.Transglutaminase are widely distributed in nature.At present,this enzyme is mainly derived from microorganisms.Transglutaminase are mainly used in the food industry to improve the protein structure of foods.It has also been reported to be associated with human diseases(including tumors,autoimmune diseases,skin diseases,neurodegenerative diseases,etc).However,the role of this enzyme in B.cinerea pathogenicity has not been reported.Expression of BcGP was induced during B.cinerea-host interaction.Thereby,we studied the role of BcGP in B.cinerea during it’s growth and it’s function in the course of interaction with ginseng.We obtained the following results:1.The BcGP gene was cloned and sequenced.The ORF of BcGP gene is about 781 bp which contains three exons with two introns.This gene encodes 226 amino acids.Protein blast analysis observed five domains,which highly conserved in evolution.2.The BcGP knockout mutant ΔBcGP and complementary transformant ΔBcGP/GP were obtained by Agrobacterium tumefaciens mediated transformation(ATMT).The knockout mutant and complementary transformant were confirmed by PCR and qRT-PCR.3.The colony growth of ΔBcGP was faster than that of the wild type B05.10.ΔBcGP was more sensitive to salt stress.The sensitive to Congo red(CR)and Sodium dodecyl sulfate(SDS)are not significantly changed.However,ΔBcGP was more tolerant to hydrogen peroxide than B05.10.4.ΔBcGP was more pathogenic to the leaves of Panax ginseng,Arabidopsis thaliana and Phaseolus vulgaris than B05.10,indicating BcGP negatively regulated B.cinerea pathogenicity.