| Salinization is a significant abiotic stress that has a severely harmful effect on China's resources and environment. Salinity is an important abiotic stresses factor, it impacts the life cycle and many biological functions of plants, such as limits the growth and development, protein-synthesis, photosynthesis and metabolic activity. Halophyte was attracted the researchers' much attention, because it has the characteristics such as rich saline-alkali resistant gene pool and it can be grow well on saline-alkali soil to improve soil situation.Halogeton glomeratus is an annual halophytes and belongs to Halogeton(family Chenopodiaceae), it is mainly distributed in the most northwest provinces, Inner Mongolia and west of northeast in China. Halogeton glomeratus has high succulence of branched stems and leaves on ground, and has strong ability of keeping soil and water, fixing-sand, drought and salt tolerance. Halogeton glomeratus is tolerant to drought, saline-alkali environment, high temperature and other harsh environment. Halogeton glomeratus is a good wild resource to obtain salt-tolerant gene and palys a decisive role in improving soil and environment in desert and saline soil. The main contents and results of this study are as follows:1. Based on the results of transcriptome sequencing in Halogeton glomeratus,we found the Unigene7547 gene that has higher expression, and designed primers to clone the full sequences of Unigene7547. The full-length sequence has 1306 bp and contains 1218 bp in its open reading frame, and codes 405 amino acids. The protein encoded by Unigene7547 is acidic protein and unstable, and the molecular weight is 45.16 k Da.2. The reliability of protein encoded by Unigene7547 that was localized in cell nucleus, mitochondrial matrix, lysosome and microbodies can be reached to 0.600, 0.100, 0.100 and 0.018, respectively. The protein is hydrophilic and has no transmembrane domain. The protein is non-secreted protein, because there is no signal peptide, which explains this protein just palys role in intracellular. Besides that, we found the Thr, Ser and Tyr has 8, 8 and 5 phosphorylation sites in Unigene7547 protein, respectively. These phosphorylation sites might affect cell physiology and regulatory modification. The random coil, extended-chain, ? helix and ?-corner was 37.78%, 28.89%, 23.46% and 9.88% in secondary structure, respectively. After BLAST homology comparison, there is no sequence was matched with the full sequence of c DNA of Unigene7547 gene, and the gene was first discovered, therefore, we presumed this gene is a new one. From the above analysis, we just made some superficial research on the biologic characteristics of the protein encoded by Unigene7547, however, these data provides a theoretical basis for the next study, and some special functions need to be further study.3. Using conventional molecular cloning techniques, we designed specific primers that contain Xba I and Sma I restriction sites, and used these primers to digest Unigene7547 gene and p BI121 plasmid of plant expression vector, then was linked up these two snippets by T4 DNA Ligase to construct the p BI-Unigene7547 of plant expression vector. The p BI-Unigene7547 was injected agrobacterium and mediated the tobacco leaves to obtain the resistant plants. After PCR amplification, the results of electrophoresis showed preliminary judgement that the Unigene7547 gene was transferred into the tobacco and expressed successfully, it provided assistance for the verification of gene-related functions, and also it is ready for the application in cultivated crops salt tolerance. |
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