Cloning, Molecular Markers Development And Function Analysis Of The Gene TaOSCA1.4 In Common Wheat

Common Wheat is a kind of important food crops, it is playing an important role for easing the pressure about growing world population.In the process of exploring a new method for cloning gene, we got a drought-resistant gene according the molecular marker SWES1079,developed a function marker and association analysis with important agronomic traits. The main results were as follows:The gDNA and cDNA sequence was obtained base on t he marker SWES1079 take advantage of wheat variety tainong 18. The gene with At OSCA1.8 in Arabidopsis, OsOSCA1.4 in Rice belongs to the same gene family through the NCBI search, so we named the gene is TaOSCA1.4 temporarily. Through sequence alignment, The three TaOSCA1.4 gDNA sequences have the same structures and all contains 11 exons and 10 i ntrons. We designd three pairs specific primers according to the specific sites in genome, and the gene were mapped on the homologous group 1 in wheat by Chinese spring nullisomic-tetrasomic lines. The sequence length of TaOSCA1.4-1A is 5142 bp, contains a 2418 bp open reading frame which encoding 805 amino acids. The sequence length of TaOSCA1.4-1B is 4975 bp, contains a 2412 bp open reading frame which encoding 803 amino acids. And the sequence length of TaOSCA1.4-1D is 4961 bp, contains a 2409 bp open reading frame which encoding 802 amino acids.Through analyzing the bioinformatics TaOSCA1.4-1B, we get the protein molecular weight was 93.3 kD, the isoelectric point is 9.16, there are many secondary structures contain a large number of random curl, alpha helix, and a small amount of beta Angle and chain extension; the leucine is richest is 87,and the followed by valine is 61. The amino acid sequence has two conserved domains, such as RSN1 TM, DUF4463 and DUF221; there are potential phosphorylation sites, contains 18 Ser, 7 Thr and 5 Tys; For the predict of proteins across the membrane that the amino acid sequence contains eight transmembrane regions. We make a Cluster analysis and find that the OSCA has three subclass.The polymorphism analysis about the sequence of 6 wheat varieties,we find that the gene contained more SNPs and InDel differences. The gDNA of TaOSCA1.4-1A contains 15 SNP loci and 3 InDel site, The nucleotide diversities formed; The TaOSCA1.4-1B contains four SNPS loci and InDel site, and also formed two haplotypes. But There is no difference in sequence of TaOSCA1.4-1D.This SNP in the position of 4253 bp in TaOSCA1.4-1B was transformed to CAPS marker. Using molecular data of the ‘Tainong 18×Linmai 6’ RIL population, TaOSCA1.4-1B was located in chromosome 1B where flanked by markers S-1036504 and S-1024654 with a genetic distance of 1.09 cM and 0.78 cM.We get the association analysis about the function marker with agronomic trait,the result is that the total number of spikelets(TSS) in 10 environment reached extremely significant difference(p≤0.01) between the haploid Hap-1B-A and Hap-1B-C. In 1 environment, we detected significant differences(p≤0.05). We also detected that the Plant high(PH) in 13 environment reached extremely significant difference(p≤0.01) between the haploid Hap-1B-A and Hap-1B-C, in 1 environment, we detected significant differences(p≤0.05).Another, They were also differences for TSSS, GNS and GYA at p≤0.05 level.