Cloning ZmLysM Receptor Genes Of Zea Mays And Functional Analysis

Plants and micro-organisms in the long co-evolutionary process,gradually formed a highly complex interaction identification mechanism,the cell surface recognition receptors recognize conserved pathogen-associated molecular patterns,and activation of downstream signaling pathways key functional proteins and then induce symbiotic or defense reaction.Recent studies have found that,LysM domain protein is an important plant pattern recognition receptors,it plays an extremely important role in the interaction of plants and fungi.To further define how LysM receptor gene specifically recognizes different ligands induce symbiotic or defensive signals downstream role.In this study,reported in trefoil and alfalfa known LysM gene as a template,combining bioinformatics methods for maize(B73)to compare whole genome,combined with software associated organisms domain genes to predict,screening of maize or adjust symbiotic defense LysM candidate genes.With RT-PCR technology from plant roots cloned candidate genes,named ZmLysM3 and ZmLysM6.Using real-time quantitative PCR were analyzed temporal and spatial expression,the results show that the two LysM gene was expressed in different tissues,especially the roots with a higher level of transcription.Different inducible factor such as bacteria peptidoglycan,LPS,flagellin flg22,fungal chitin,AM Fungi maize roots,and found ZmLysM3 and ZmLysM6 transcript levels can be significantly increased.It can be seen that the gene may be involved in bacterial or fungal pathogen secreted ligand recognition.Structure forecast indicates that LysM,a transmembrane domain,and intracellular kinase domain compose the ZmLysM3,in contrast,ZmLysM6 lack the intracellular kinase domain.To clarify the position of two LysM proteins in the cell,ZmLysM3 and ZmLysM6 genes are ligated into an expression vector pCAMBIA1305.1-GFP,the Agrobacterium-mediated transient expression in tobacco leaves,the sub-cellular localization of gene testing,results,these two proteins are located on the cell membrane.Yeast two-hybrid experiments show,ZmLysM3 and ZmLysM6 protein extracellular domain by forming a heterodimer to recognize the extracellular ligand-induced signal transduction in turn downstream.To further define the signaling mechanisms mediated by both,the study which were transferred into Arabidopsis and co cerk1-2(Arabidopsis lacking chitin texture system known mutant)for functional complementation mutant feature.Leaves of transgenic plants induce fungal treatment 72 h,withered by the degree of inhibition of plant leaves and the result leaves hyphae seen,and simultaneously turn ZmLysM3 transgenic Arabidopsis ZmLysM6 significantly enhancedplant disease resistance,but only turn into a LysM inhibitory effect on the fungus it is not obvious.The results described above,in theory,will explain how corn LysM gene regulation studies of plant resistance mechanisms to provide new content and modes.In practice,the resistant varieties by fusing the extracellular receptor kinase domain,construction of chimeric receptors,increase resistance to susceptible plants.Therefore,the study carried out LysM receptor kinase has produced on the necessity for hybrid breeding female,increase resistance to future generations of great significance.