Isolation And Functional Identification Of Salt Tolerance Genes From Ceriops Tagal

Land salinization is one of the abiotic stress that restrict the growth, development and production of crops. In response to the increasingly serious soil salinization at present, the study mainly focuses on the bioinformatics and molecular biology on the root system of Ceriops tagal(mangrove plants)(500mmol·L-1) under salt stress at different time point. Selecting different time point for the screening of differential gene, transcriptome and the sequencing of Digital Gene Expression Profiling on the root system sample were carried out at different time point by using the newest high throughput sequencing RNA-seq.Afterwards, forty genes were selected as candidate for semiquantitative and real-time verification according to the expression profiles analysis; then eight genes were proceeded with vector construction as well as the analysis on the function of these genes through the transgenosis study on Arabidopsis. The results were as follows:(1)The expression of total protein in Ceriops tagal was determined under different level of salt stress. The result revealed that the content of protein in Ceriops tagal declined prior to the increase over time, suggesting that the numbers of protein in Ceriops tagal could be adjusted by salt stress.(2) The total RNA processed at seven different time point (500mmol·L-1NaCl), was extracted from Ceriops tagal, proceeded with illumine sequencing and biological information analysis related with databases, such as COG and GO. Then according to the information supplied by transcriptome overall length of CDs sequence from part of differential expression gene in Ceriops tagal was discovered.(3) In accord with the expression profiles analysis, forty genes with relative high difference were screened to design primer, and verified by RT-PCR and real-time PCR, showing75per cent of consistence in the result.(4) Semiquantitative RT-PCR coupled with RACE technology was proceeded and eight genes with overall length of CDs were isolated, including Mn-SOD.(5) Utilizing LIC-cloning method, eight screened genes relevant to salt tolerance were built into the expression vector pJG053of the plant, converted into agrobacterium GV3010, and the T2generation transgenic plant with stable inheritance was obtain after Arabidopsis was infected by the method of floral dip.(6) Salt tolerance experiment was carried out on T2generation transgenic plant, including genes of CL2528, CL3363, CL3369and CL3122. The result indicated that the growth of transgenic Arabidopsis by genes of CL2528and CL3363appeared to be more preferable than that of the plant without transgenosis, suggesting the presence of salt tolerance related function of the two genes.