Comparative Analysis Of Telomerase Activities Related To Tree Age And Abiotic Stress Tolerance

Telomeric repeat amplication protocol(TRAP) is a common method for detecting telomerase activity. However, it is not completely suitable for the detection of telomerase activity in tree plants. So, we modified the TRAP assay and made it stable and high sensitivity for woody plant material, whose development differs in fundamental ways from that of hrebaceous plants. According to the previous research, PEG8000was added to the telomerase extracts at a final concentration of10%. Comparative telomerase assay results conformed that PEG8000may reduced the concentration of inhibitors to Taq polymerase or telomerase that are commonly found in crude extracts. TS was choosed as the primer and100ng of protein sample as the template concentration for PCR. Fourteen μl aliquot of each PCR product was applied to a12%non-denaturing polyacrylamide gel and electrophoresed. The gels were stained using the SYBR Green I and photographed under ultraviolet light using an electronic gel documentation system.In order to determine relations of telomerase activity to tree age and leaf position in Pinus tabulaeformis, Ginkgo biloba, Catalpabungei C.A.Mey, and Sophora japonica Linn, we carried out the analyses of TRAP. No significant difference of telomerase activity was found in different ages of trees. But the hightest telomerase activity was found in the top position leaves of the tree. The telomerase avtivity decreased with the position of leaf drawdowning.It has been shown previously that apoptosis of plant cells was induced by salt stress. We treated the Populus euphratica callus and Ammopiptanthus mongolicus suspension cultured cells with salt in different dense. Experiment reported here demonstrate that cell death can be prevented if the salt treatment in lower dense, and it is irreversible that the cell treated with salt in the higher dense. At the same time, we analysed protein conservation and phylogenetics of TERT, and determined that TERT is a highest conservative protein. So we designed the primer and clone the TERT in Ammopiptanthus mongolicus. In the present, we have gained a part of TERT sequence of Ammopiptanthus mongolicus.