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Cloning of Gene(s) Induced by Salt Stress & Analysis of Its Function in Wild Barley (Hordeum Brevisubulatum (Trink.) Link.)

Author: QiXiaoQiang
Tutor: WengYueJin
School: Chinese Academy of Agricultural Sciences
Course: Biochemistry and Molecular Biology
Keywords: Wild barley Salt Tolerance SSH RACE cDNAs
CLC: S512.3
Type: Master's thesis
Year: 2003
Downloads: 263
Quote: 7
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Abstract


The purpose of this test is to find and plant salt tolerance genes in order to develop our own intellectual property resilience genetic resources, the use of genetic engineering means cultivating new varieties of salt. In this study, the wild two-rowed short awn barley as test materials, the use of SSH (suppression subtractive hybridization) methods to compare the normal culture (CK) and 430mM NaCl salt solution stress treatment 24hrs (T) gene transcription level differences cloned differentially expressed cDNA fragments 449, differentially expressed cDNA library was constructed. Reverse Northern Bloting, 40 positive clones were identified, selecting differentially expressed sequenced EST 18, removing one repeat obtained 17 EST sequencing results, the selection of 1 for RACE, finally to obtain full-length cDNA Gene BST16. On this basis, the use of NCBI, TIGR and other public databases bioinformatics analysis and functional predictions. The results showed that plant salt tolerance of plants involved in many physiological and biochemical pathway by multiple gene regulation. The institute's 17 EST fragments obtained through analysis and function prediction, which Bst3, Bst9, Bst18 involves barley rbcL gene, known to induce the gene and light, cold and osmotic stress related; Bst1, Bst6, Bst17, Bst22 with barley specifically expressed under salt stress stress protein genes: Bst13, Bst19 and bad environment, helps to maintain the activity of molecular chaperone protein related protein coding genes; Bst4 membrane protein coding genes, prediction of protein kinase activity; Bst5 mitochondrial genome NADH dehydrogenase gene fragments; Bst8 wheat plants with ethanolamine phosphate methyltransferase gene homology; Bst10 with one in rice and Arabidopsis have been found ATP-dependent protease ATP-binding subunit gene highly homologous to the gene encoding protein is likely to be a heat shock protein; Bst11 of cAMP cGMP-dependent protein kinase gene, may be involved in signal transduction: Bst12 rice isocitrate lyase gene has no homology sex; Bst20 based on an analysis of the translation initiation factor-related protein-coding genes, and protein synthesis. Total length gene BST16 601bp, its read reading frame analysis found that 93-515 nucleotides from a sequence that contains the initial password and the stop codon, encoding a 140 amino acid polypeptide. The full-length gene sequences in GenBank and TIGR database for comparison, a sequence encoding the barley PS Ⅱ 10kD protein 469bp fragment of up to 90% homology with the Arabidopsis 516bp PS Ⅱ polypeptide coding sequence with 65% homology , with wheat under drought stress period of 466bp cDNA of 94% homology. Reading frame of the gene encoding the protein functional sites and structural analysis of functional domains, also show that the gene coding for the PS Ⅱ 10kD protein gene, the amino acid sequence of 48 abstract Mateo position contains a conserved photosystem * 10kD protein domain PSbR, the protein has a proximal end ape conservative short-chain dehydrogenase / reductase family signature sequence ADHSHORT (Short-chain dehydrogenases / reductases family signature), located 94-122 bits, indicating that the protein might have reductase activity . Taken together, these analyzes can be initially identified as the gene can be stress-induced photosynthetic systems * component 10kD polypeptide coding genes, its function and crops under salt stress is closely related to the efficiency of photosynthesis.

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CLC: > Agricultural Sciences > Crop > Cereal crops > Wheat > Barley
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