The Poplar PPDK And PDRP Gene Cloning And Functional Verification

In nature, higher plants can be divided into three categories of photosynthetic types:C3, C4and CAM. C4and CAM plants,under stress conditions over a long period of domestication, evolved from C3plants. Compared with C3plants, C4plants have higher photosynthetic rate and nutrition and water use efficiency at high light intensity, high temperatures and low concentrations of CO2and drought conditions. However, C3plants have the corresponding homologous genes that involved in photosynthesis in C4plants. To further study the mechanism of key enzyme PPDK (Pyruvate orthophosphate dikinase) in C3plants, we found that PPDK phosphorylation and its phosphorylation sites(Thr581) by use of mass spectrometry technology in C3plants of Populus(Populus simonii×P. nigra);then the corresponding C4photosynthesis key enzyme PPDK gene and its regulatory gene PDRP are cloned from the Material. Literature retrieval and bioinformatics analysis show that The two species PPDK protein have conservative pyruvate kinase domain, PEP binding domain and transfering domain, while both PDRP protein have ATP-binding domain. And, further we prove that PPDK can be phosphorylated in vitro and its phosphorylation sites is the Thr581, which is very similar to previously reported C4plant maize PPDK phosphorylation by use the prokaryotic expression and purification of the PPDK and PDRP protein. In addition, the online microarray data analysis shows the distribution of PPDK and PDRP in plants, as well as the level of expression in different stress treatment in Arabidopsis. This study provides some clues and basis to further explore the mechanism of C4photosynthesis key enzyme in C3plants, as well as molecular genetics and breeding of poplar and other trees.