The Subcelluar Localization And Functional Characterization Of Triticum Asetivum L. Glycogen Synthase Kinase 1 (TaGSK1)

From the wheat salt tolerance mutant RH8706-49, our lab had cloned the gene TaGSK1. To construct the binary express vector pBIN-Ta-GFP, we tagged the gene TaGSKl at the carboxy terminal end with green fluorescence protein (GFP). With the control vector pBINGFP4, we transferred the two vectors into agrobactrium GV3101 and got the positive clones with the screening of kanamycin. Using the method of floral dip with the screening of kanamycin, we got the positive transgentic Arabidopsis thaliane lines ATaGFP and AGFP. We plant the T2 seeds of ATaGFP and AGFP onto MS medium (supplemented with kanamycin). After 5 days, view the root cells of young plants under confocal. We found that the countral experiment showed that the green fluorescence existed at the nucleus and membranes of AGFP line. But the most green fluorescence exist in the cytoplasm of ATaGFP line root cell. The results demonstrated that the protein TaGSKl exists in the cytoplasm of plant cell.Planted the seeds of ATaGFPx AGFP and Ac onto MS medium supplemented with different concentration of NaCl (70 mmol/L NaCl, 120 mmol/L NaCl). After seeding 7 days; measured the length of taproots of young Arabidopsis. With the analysis of SSR(shotest significant ranges), we found that the average root length of ATaGFP AGFP and Ac on 70mmol/L NaCl MS medium were 8. 509cnu 6. 722cm andl. 493cm. The difference between ATaGFP and AGFP was statistically significant (p<0. 01). The value between ATaGFP and AGFP was significant (p<0. 05) which demonstrated that TaGSKl promote the root of transgenic Arabidopsis growth. On the 120mmol/L NaCl MS medium, the average root length of ATaGFP AGFP and Ac were 4. 600cm, 2. 532cm and 0. 719cm. Between the three lines the different were statistically significant (p<0. 01). After seeding on salt MS medium (70 mmol/L NaCl, 120 mmol/L NaCl) 9 days, counted the number of lateral roots. On the 70 mmol/L NaCl MS medium , the number of lateral root of ATaGFP AC and AGFP were 1. 737, 0. 673 and 0. 034. On the 120 mmol/L NaCl MS medium, the number were 0. 976, 0. 289 and 0. 000. The differences between these lines were statistically significant (p<0. 01). The result primarily demonstrated that TaGSKl promote the tolerance of transgenic Arabidopsis to NaCl. When we were doing the experiment of subcellular localization, we fount that there were lots of TaGSKl existed on the root tip and thefoundation of lateral roots. We presumed that the existence of TaGSKl on the root tip and the foundation of lateral roots is one of the main reason to promot the taproot growth and the formation of lateral roots.Plant the seeds of ATaGFP and AGFP in row onto the MS medium. After 6 days treated the young plants with CBW(1. 3 mol/L sucrose) . We found that the root cells of AGFP had plasmolyzed, but the root cells of ATaGFP did not. When we treated with 2. 0 mol/L sucrose on ATaGFP, the cells plasmolyzed. Treated the cell of onion pellicle with different CBW, we got the same result as above. The results demonstrated that TaGSKl could promote the tolerance of plant cells to ostomic stress.