The Primary Study Of The Wheat Fructose 1,6-Bisphoshate Aldolase From Albinism Line

Wheat albinism line is a natural mutant from the parent dwarf wheat Aibian 1, which is sensitive to the low temperature, and became a stable heredity material. In winter and early spring, the leaves of this line turn to albinism. Up to now, many study about the wheat albinism line's genetics, cytology, physiology and biochemistry,chloroplast development,et al.thave been carried,and have acquired a lot of important results.This study is on the basis of Ph.D Hou Dian yun on the study of contrast of chloroplast protein about wheat albinism line and Aibian 1 at low temperature.We selected the fructose-1,6-bisphosphate aldolase as the object of study,the enzyme play an important role in the Calvin cycle,which participate in the carbon and nitrogen metabolic of chloroplast. There has reported that the enzyme have a positive effect on tobacco in salt stress. The study from gene sequence comparison, variation trend of the enzyme,transient express carriers transform protoplast and real-time pcr et al.to make a study about express speciticity the wheat albinism line fructose-1,6-bisphosphate aldolase at low temperature,and the major results as follow:1. The fructose-1,6-bisphosphate aldolase gene genomic DNA and cDNA have been cloned from albinism line and Aibian 1. After the gene sequence comparison, there were three different amino acid mutant compared with the publiced gene sequence (FJ625793).There were no difference on genomic DNA and cDNA between wheat albinism line and Aibian 1.2. In room temperature, the enzyme activities of two materials are samely. After low temperature treat, the variation trend result of the enzyme activity show that two materials have a difference model. Aibian 1 have a obvious periodicity change, the variation trend like up-down-up, and is relatively stable. At the fourth day after cold treat, the enzyme activity of albinism line is the most high, then it decrease obviously.3. We established the system of protoplast transform system, and succeed transfor the transient express vector of fructose-1,6-bisphosphate aldolase gene with eGFP to the leeks protoplast. We observed the expression of eGFP in cytoplasm.4.The real-time-qutantity pcr were used to analysis the CpFBA gene expression change in low temperature. The results show that the gene expression level of albinism line is same as the Aibian 1 aldolase at room temperature. At the fourth day of low temperature , the expression of albinism line decreased distinctly, but the express level of Aibian 1 increase rapidly and it's higher than albinism line. This show that the albinism is affect by low temperature.During the low temperature conditions later, the expression level of Aibian 1 tend to stability,but wheat albinism line is lower at first,then,it's ascend,at within the 20 days tend to little variation,however. But at the 34th day, the gene expresssion of albinism line rised up to the most high lever.The final result show that the alodlase is affected by low temperature.