The Roots Salt-induced Genes HvSR1 Identification In Barley And Construction Of Vector Specially Used In Gene Function Analysis

Salt stress can significantly affect crop yield and quality, and roots play key roles in plant responses to salt stress. Therefore, identification of salt-induced and root preferentially expressed genes for molecular breeding, is one of important ways for genetic improvement of crop salt tolerance. Barley is an important model crop with genome sequenced fully. As sequencing technology develops, barley transcriptomic data grow rapidly. Hence, exploration and validation of relevant databases becomes more and more urgent and crucial for knowledge enhancement and phenotype clarification. In this study, transcriptomic data from 48 samples in eight experiments were analyzed by Genevestigator online bioinformatic platform, and four salt-induced and root preferentially expressed genes were identified. Gene expression patterns were verified in different tissues and salt treatments using barley cultivar, Morex. And vectors for functional analysis of barley genes were constructed.The results showed that the gene corresponding to probe Hv.1961 can be significantly upregulated by salt stress and redundantly expressed in barley roots, and designated as Hv SR1 gene(Salt Responsive Gene 1). Full-length coding region of 1377 nucleotides encodes 458 amino acids containing two conserved domains. DUF247 belongs to unknown proteins in plants. Csb2_I-U may be related to CRISPR / CAS system proteins.Gene expression patterns of HvSR1 gene from bioinformatic analysis was validated through different experiments. The results showed Hv SR1 gene highly expressed in barley roots with the highest level in mature roots, while less or no RNA detected in other organs. So Hv SR1 gene can highly express in barley roots. Another experiment revealed that expression of Hv SR1 gene can be induced redundantly by salt treatment.Two vectors for functional analysis of barley genes were successfully constructed.Selective marker gene NPTII under control of a strong maize promoter Ubiquitin in p EGFP-UN. As a enhance trap vector, GFP(green fluorescent protein) can be detected once T-DNA inserts into an enhancer area of gene promoter. p BA9 N is a small size vector(6478 bp) with kanamycin resistance marker gene under control of maize Ubiquitin promoter. These vectors build a solid foundation for establishing T-DNA mutant population in barley.