Functional Study Of Deacetylase ZmHDA101 In Maize Disease Resistance

Protein acetylation is a ubiquitous reversible protein modification mode in eukaryotic organisms.It is jointly acted by histone acetyltransferases(HATS)and histone deacetyltransferases(HDACs),which dynamically regulate the acetylation level of plants according to the growth and development needs of plants and the changes of the external living environment,thereby regulating the expression of different genes to maintain the survival needs of plants.In this study,the genes of the maize histone deacetyltransferase(HDAC)family were analyzed by bioinformatics,and the maize ZmHDA101 gene homologous to Arabidopsis AtHDA19 was obtained,with the gene code of GRMZM2G172883;Real-time was used.PCR technology was used to analyze the expression of HDAC family genes in Maize in response to salicylic acid and jasmonic acid hormone treatment;subcellular localization analysis of ZmHDA101 gene was carried out by fusion expression of GFP vector with ZmHDA101;Fusarium graminearum,Exserohilum turcicum,Bipolarismaydis(NisikadoetMiyake)Shoem,Helminthosporium carbonum,Curvularia lunata Boedijn were monitored by monitoring the overexpression plants of ZmHDA101 in maize.The function of ZmHDA101 gene in maize disease resistance was analyzed.The interaction between ZmHDA101 and ZmTPL/TPRs was studied by yeast two-hybrid technology,which laid a foundation for elucidating the function and mechanism of maize HDAC family genes in maize resistance to biotic and abiotic stresses.The main results are as follows:1.Bioinformatics analysis of HDAC family genes in maize.The phylogenetic and structural domains of maize HDAC family genes were analyzed,and it was found that maize HDAC family genes can be divided into five subfamilies(Class Ⅰ,Class Ⅱ,Class Ⅲ,SRT and HD2),of which Class Ⅰ,Class Ⅱ,Class Ⅲ subfamilies all contain the domain specific to deacetyltransferase HDAC,and ZmHDA101 belongs to Class Ⅰ subfamily.Using bioinformatics methods,the expression of HDAC family genes in Maize under tissue-specific,abiotic and biotic stresses was analyzed,and it was found that the expression levels of HDAC family genes in Maize at different developmental stages,different tissues and in response to different abiotic and biotic stresses showed significant differences.The expression of ZmHDA101 gene was stable in different parts of the plant,and was significantly up-regulated under high temperature and high salt stress.It is speculated that maize HDAC family genes are involved in Maize growth,development and response to abiotic and biotic stresses.2.Analysis of expression pattern of maize HDAC family genes in response to salicylic acid and jasmonic acid hormones in maize.Real-time PCR was used to detect the expression of maize HDAC family genes in response to salicylic acid and jasmonic acid hormones in maize.The results showed that maize HDAC family genes showed different rules in response to salicylic acid and jasmonic acid treatment.ZmHDA101 showed an induced expression trend under jasmonic acid hormone treatment and a downregulated expression trend under salicylic acid treatment,suggesting its involvement in jasmonic acid and salicylic acid signal transduction pathways.3.Subcellular localization analysis of ZmHDA101.Using Getway technology,the vector 103-ZmHDA101 was constructed to express ZmHDA101 fused with GFP.103-ZmHDA101 was transiently expressed in tobacco leaf cells by Agrobacterium transformation.The green fluorescence signal of GFP was observed under laser confocal microscopy,and the localization of ZmHDA101 gene in the nucleus and plasma membrane was determined.4.Function of ZmHDA101 in Maize Disease Resistance.The ZmHDA101 overexpressing plants were inoculated with Fusarium graminearum,Exserohilum turcicum,Bipolarismaydis(NisikadoetMiyake)Shoem,Helminthosporium carbonum and Curvularia lunata Boedijn for disease resistance identification.The results showed that compared with maize inbred line B73,ZmHDA101 overexpressing plants showed obvious resistance to the tested pathogens inoculated in the experiment,indicating that ZmHDA101 plays an important role in the process of maize disease resistance.Real-time PCR was used to detect the expression level of PR gene in maize inbred lines B73 and ZmHDA101 overexpressing plants.The results showed that the expression of PR gene was significantly lower in ZmHDA101 overexpressing plants that were not inoculated and inoculated with Fusarium graminearum for 48 h than in inbred line B73,indicating that the overexpression of ZmHDA101 affected the expression of PR gene,further illustrating that ZmHDA101 plays an important role in maize resistance to pathogen infection.5.A yeast two-hybrid interaction study between ZmHDA101 and AtTPL/TPRs.The interaction between ZmHDA101 and AtTPL/TPRs was studied by yeast two-hybrid technique.The results showed that the co-transformed yeast AD-ZmHDA101 combined with BD-AtTPL,BD-AtTPR1,BD-AtTPR2,BD-AtTPR3,BD-AtTPR4 and the combination of BD-ZmHDA101 with AD-AtTPL,AD-AtTPR1,AD-AtTPR2,AD-AtTPR3,AD-AtTPR4 all grew well on the yeast three-deficiency medium,indicating that ZmHDA101 and AtTPL/TPRs family genes could interact directly in yeast cells.