Analysis And Functional Verification Of Thil Gene Family In Barley

The Thionin-like gene(Thil)has been well known for its antiviral activity in plant defense system in Arabidopsis thaliana,but there were very few studies in barley and other crops.The Thil gene family of barley was identified and analyzed by sequence composition,gene structure,chromosome distribution and protein function prediction,and based on phylogenetic tree,expression heat map,gene promoter elements and subcellular localization.q RT-PCR was used to analyze the expression pattern,observe the expression changes under stress conditions,conduct preliminary functional analysis,and further conduct gene haplotype analysis with 1000 core germplasm resources.The main conclusions are as follows:1.Using barley genome data,the members of Thil gene family were analyzed at the whole genome level of barley.A total of 7 Thil genes,ranging in length from 227-414 bp,were identified on chromosomes 1H,2H,4H,5H and 6H of barley,respectively.Phylogenetic tree analysis of Thil protein construction in barley,Arabidopsis thaliana and rice revealed that Thil proteins in the three species could be divided into four groups.Thil gene family members are mainly expressed in leaves,embryonic tissues of four days of development,young roots and old roots.2.Gene structure analysis showed that Thil1,Thil2,Thil4 and Thil7 have transmembrane domains,while Thil3,Thil5 and Thil6 have no transmembrane domains.Except for Thil4,the sequences of other family members are relatively conservative,and all members are There are signal peptides.This family interacts with most amino acids(100% ligand probability),and has the highest probability of becoming a real ligand with chloride ion,citrate anion,phosphate ion and sulfate ion,etc.3.Using q RT-PCR to analyze the expression patterns of Thil genes under salt stress,exogenous ABA stress and mechanical damage stress,it was found that all Thil genes can respond to these three stresses,with different expression patterns.same.4.The Subcellular localization results showed that Thil1 and Thil2,Thil3 and Thil7,and Thil6 were located in chloroplasts,except Thil4 and Thil5,which were not cloned successfully.5.The haplotype analysis of barley found that the variation of Thil2 gene was the most,and its expression level under stress was also at a high level,so this gene has a certain reference value for breeding excellent varieties.