Genetic Analysis And Ecological Association Of HinA Genes In Wild Barley

According to the conserved regions of barely HinA gene,the gene-specific primers were designed to amplify the full gene sequences from 127 wild barley accessions belong to 19 populations from Iran,Israel and Turkey.Based on the results of multiple sequence alignments of HinA gene from 127 wild barley accessions,a number of single nucleotide polymorphisms(SNPs) were detected.The relationships between the SNP variations of HinA locus and the ecological factors were investigated.And the deduced amino acid(AA) sequences and their second structures of HinA protein variations between cultivar and wild barleys were discussed.The following results were obtained.1.The analysis of 127 HinA sequences of wild barley was carried out.A total of SNPs (one SNP per 29.6bp) were found in coding and non-coding regains,and ratio of base substitution between transition and transversion was 0.706 to 0.294,which was very similar with other barley genes.Fifteen haplotypes were found in the coding regain of HinA gene in wild barley,and were scattered in the populations.2.The variation of genetic diversity(He) of HinA genes was similar as those of Shannon's information index(Ho) and polymorphic per population(P).About 2/3 genetic variations of HinA genes were presented within population,while about 1/3 genetic variations were observed between populations.Broad gene flow(Nm=3.31) and lower genetic variation(Gst=0.0702) were detected,and there was not significant correlation between genetic distance and geographical distance between populations(r=-0.005, p=0.478).3.The results of spearman rank correlations(r_s) analysis showed that only correlations of between P,He and Ho of Israel and mean humidity at 14:00 were close to significant(p=0.071;p=0.052;p=0.052).The result of spearman rank correlations between polymorphic of SNP and ecological factors showed that 81.7 percent SNPs were significant correlated with ecological factors.Among ecological factors,the number of SNPs affected by average humidity at 14:00 was the largest(25),while that affected by longitude was the smallest(11),indicating that the connection between average humidity at 14:00 and change of HinA was the tightest,while that between longitude and change of HinA was the loosest.4 The AA sequences of HinA genes were classed into groupⅠand groupⅡbased on the neighbor-joining analysis of MEGA3.1.GroupⅡincluded three subgroups(i.e.Ⅱ-1,Ⅱ-2,andⅡ-3).Most of HinA genes were clustered into groupⅠand groupⅡ-3.On the basic of AA(Ile/Thr) at 146,cultivated barley and wild barley could be distinguished,while groupⅠandⅡcould be distinct with help of the 71^th site and the 78^th site in wild barley. According to AA sequence and protein second structure,the result showed that there were 3α-helixes 5β-pleated sheets,and three diversity-regains(30-40,70-80 and 140-150) in the HinA genes.