Functional Analysis Of SikPIP1;5 And SikPIP2;7 Gene Of Saussurea Involucrata Kar.et Kir

Water is an ubiquitous and indispensable molecule for plant growth and development. They transport between cells is mainly mediated or by aquaporins finished film. Aquaporins are part of MIP(Membrane Intrinsic Protein) family, they are multifunctional membrane channels and facilitate the transport of water across plant cell membranes,mainly divided into seven subfamilies. PIPs are located in the plasma membrane aquaporin family of water transport and rapid plant selective membrane transport play a key role. When the plant suffered salt, drought, cold and other abiotic stresses, PIPs to quickly transport the body of water on the plant and make the regulation of intracellular osmotic pressure, reduce stress on plant damage. In this study, two aquaporin genes(Sik PIP1;5, Sik PIP2;7) cloned from Sasussured involucrata, biological information, the cellular localization and transgenic plants and yeast adversity adaptive resolution. The main findings are as follows:1 The Sasussured involucrata’s Sik PIP1;5 followed by 867 bp of open reading frame encoding 288 amino acids, The molecular weight of 30.637 KDa, p I of 9.09. The Sik PIP1;5 followed by 840 bp of open reading frame encoding 279 amino acids, The molecular weight of 29.01 KDa, p I of 9.26. The analysis of the amino acid sequence reveals that they contains the MIP family sequence SGXHXNPAVT,and the PIP family signature sequence GGGANXXXXGY and TGI/TNPARSI/FGAAI/VI/VF/YN.Two of them exhibits six predicted transmembrane helices and possesses two sets of conserved NPA motifs. Homology and sequence analysis found Sik PIP1;5 should be a member of PIP1 subfamily and the Sik PIP2;7 should be a member of PIP2.2 The transgenic tomato plants were obtained by Agrobacterium-mediated method, the callus differentiation, adventitious buds, rooting, Kan screening, PCR and RT-PCR analysis indicated that the aquaporin protein gene Sik PIP1;5and Sik PIP2;7had been integrated and transcribed into the tomato genome. Agronomic traits survey found that transgenic tomato were dwarfed, the leaves number and the number of branches increased in trans- Sik PIP1;5 tomatos, trans-Sik PIP2;7 tomatos can early flowering and fruit. Transgenic tomatos and wild-type tomatos into low temperature control, drought and Na Cl stress test, Through morphology observation and its physiological indicators(related leaf relative water content, relative conductivity, MDA content)and antioxidant enzyme activities were measured by building plants. The results show that under stress MDA content and relative conductivity were increased, but the wild-type rise significantly higher than the turn genotype; antioxidant enzymes SOD, APX, POD and CAT activity were higher than wild-type. This suggests that transform Sik PIP1;5 and Sik PIP2;7genes from Sasussured involucrata Kar can reduce the toxic effects of ROS in cells, protected the cell membrane system,and enhance the plants resistance to drought, high salt and low temperature.