Cloning And Functional Analysis Of OsSRT Gene In Rice

Histone acetylation is an important modification of proteins from yeast to plant. In growth and development, cell proliferation, damage repair. The histone acetylation is playing an important role. The deacetylation and acetylation the have the dynamic equilibrium. The key enzyme, histone acetyltransferase(HATs) and histone deacetylases(HDACs) are playing important roles. The histone deacetylases can be divided into three families, RPD3, HDA1, HD2 and SIR2 family, In different types of organisms have different functions.SIR2 family as an NAD+dependent histone deacetylase family is well known. They have some different structures on the other family members. There is no sequence homology.which also implies that, SIR2 histone deacetylase particular the functional activity. At present the studies have shown that SIR2 family has the effect on chromosome modification,change the structure of chromosomes. but for the family’s role in plant growth and development, Especially on the role of stress in plants is little known.We choise the Os SRT1 of SIR2 family in rice for study.The gene accessio n number(AK069000.1), We use the Biozol way to extract total RNA in rice.As known the sequence by NCBI and primers were designed by using the so ftware.Using the RT-PCR way to get some specific fragments. Sequencing the ORF fragments is 1453 bp, and the known sequence fragment similarity of 99%.In order to further analyze the gene in growth and development. Particularly inthe face of stress function. using Gateway kit construct into a plant expressio n vector obtained p GWB2 p GWB2-Os SRT1; Using the electroporation method p GWB2-Os SRT1 transferred to Agrobacterium; Success by identifying the expression vector into Agrobacterium tumefaciens. Using the Agrobacterium-mediated t ransfer of the carrier p GWB2-Os SRT1 the model organism Arabidopsis transge nic plants T0 resistance screening, and molecular identification.Transgenic plants proline determination:The WT, p B2, Transgene type T2-Os SRT1-2 test material in normal culture medium with MS medium for two w eeks than put them into 0,50,100,150mmol/LNa Cl stress treatment for a week.T hen determination proline content showed that T2-Os SRT1 type in 150mmol/L when the maximum degree of accumulation of proline, respectively, 1.39 and 1.47 times of the wild-type and p B2. Changes in proline content in mannitol un der obvious stress at 200mmol/L mannitol, turn genotype T2-Os SRT1-2 were 1.37 and 1.11 times the wild-type and p B2 in 10mmol/L H2O2, T2-Os SRT1-2 ar e wild-type and p B2 1.12 and 1.20 times.Determination of soluble protein transgenic plants: the same training metho ds, the concentration of Na Cl in the above treatment, the results showed that t he highest content of 100mmol/L Na Cl T2-Os SRT1-2 turn genotype soluble pro teins, which are wild-type and p B2 1.04 and 1.13 times, while 150mmol/L Na Cl, soluble protein content of the three, has declined, but the thransgene type decreased less. the same approach at 0~200mmol mannitol stress, soluble protei ncontent progressively incremented in 200mmol/L reached a maximum,respectiv ely, of wild-type and p B2 type are 1.08 times 1.10 times, while at the use of H2O2 stress, the amount of accumulated 7.5mmol/L when the maximum solubleprotein, transferrin genotypes T2-Os SRT1-2 p B2 wild type and 1.28-fold and 1.09-fold.Activity of transgenic plants CAT and POD enzymes: In these culture c onditions, the difference in 50mmol/LNa Cl processing of CAT activity in the th ree large turn genotype T2-Os SRT1-2 are wild-type and p B2 of 1.34 and 1.19 fold, at 150mmol/L treatment, larger differential activity of POD, T2-Os SRT1-2respectively 1.27 and 1.37 times of the wild-type and p B2, while in mannitol200mmol/L, the transgenic group CAT enzyme activity larger, respectively, of wild-type and type p B2 1.12 and 1.07 times in the H2O2 10mmol/L, CAT acti vity, turn genotype T2-Os SRT1-2 p B2 are wild-type and type of 1.32 and 1.25 times.