Determination Of Biochemistrical Parameters On Salt Dressed And3’-RACE Clone And Sequence Analysis Of Salt Tolerance Gene AsNHX In Alhagi Sparsifolia

Salt damage to plants. The plants absorb moisture while absorbing the excess salt ions,especially Na~+having a toxic effect on plants. Halophytes has been able to survive in the high saltenvironment, because they can be effectively absorbed excess Na~+compartmentation into thevacuole, thereby preventing the toxic effects of too much salt in the matrix ions. In thismechanism, the vacuolar membrane Na~+/H~+antiporter carrier (NHX) protein plays a moreimportant role. NHX gene is a major gene conferring salt tolerance in plants. Numerous studiesshow that when the overexpression the NHX gene can increase the salt tolerance of plants, therebyincreasing the resistance of plants to salt stress. With the deepening of the halophyte salt tolerancemechanism studies, many plant NHX genes have been cloning.Alhagi sparsifoliA is a very good biological resistance material with the characteristics ofcold resistance, drought resistance, salt resistance and wind characteristics.The seedings of Alhagi sparsifoliA were treated by0mmol/L,50mmol/L,100mmol/L and200mmol/L NaCl solusion, and determined the biochemistricao parameters changes includingMDA,SOD,POD,CAT and Pro in leaf, stem, fibrous root and taproots. The result is Salt stress,especially high concentrations of salt stress (100mmol/L and200mmol/L taproots and leaves ofSOD active role in promoting more significantly, also accompanied by POD activity risesignificantly induced, POD plays a major role on against oxidativestress. SOD, POD and CATactivity increase probably is one of the reasons about the Alhagi sparsifoliA having a strong saltresistanceIn order to study on the core sequence of the Alhagi sparsifolia’s NHX1gene, Extracted thetotal RNA from the Alhagi sparsifolia’s leaves used CTAB method, hot borate method and Trizolkit method, and determined using Trizol kit method to Extracted the total RNA from the Alhagisparsifolia’s leaves.Then designed the special primers to synthesis the1stcDNA, and amplified by PCR, gained a DNA belt about600bp to800bp, which conforms to the core sequence of theNHX1gene that had published, and then linked the cDNA to Amplified the core As-NHX by3＇-RACE, connected to Pmd19-T vector and transfer to the DH5α cell, sequenced the positive clone.The sequence result shows the length of the As-NHX is1484bp, encode was470amino acid. Thenucleotide sequence homology is96%,89%and89%, The amino acid sequence homology is97%,93%and90%by compared the sequence with the sequence of the NHX1between SaNHX1(Salsola affinis), HaNHX1(Haloxylon ammodendron) and Kf NHX1(Kalidium foliatum) reportedin NCBI by blast soft.