Cloning And Function Analysis Of Phosphate Transporter In Brachypodium Distachyon

Phosphorus is not only an important component of nucleic acid, phospholipids and ATP in plant, but also an important nutrient elements for plant growth.As the energy transfer material, it plays important roles in activating protein , regulating metabolism process and adjusting plant growth and resistance. Usually, effective phosphorus in soil that plant can uptake is not high for meeting the need of plant growth. Phosphate transporter plays an important role in absorbing the phosphorus, especially the Pht1 gene family, which were almost high affinity phosphate transporters, and mainly expressed in roots and regulated by phosphorus deficiency.In this study, we predicted phosphate transporters in Brachypodium Distachyon by Bioinformatic method.The gDNA and cDNA sequences were got through homology cloning. According to the bioinformatic analysis to these genes, the members of Pht1gene family in Brachypodium were predicted and tissue expression pattern was investigated with quantitative realtime PCR .Some genes were conducted in Arabidopsis and Yeast carrier for function verification.The detailed results were as follows:18 phosphate transporters were predicted in Brachypodium distachyon. According to their locations in chromosome, they were named Bd1g00700,Bd1g42610 , Bd1g52590 , Bd1g67330 , Bd1g75020, Bd1g75030, Bd1g76010, Bd2g10810, Bd2g45520, Bd3g12590, Bd3g27680, Bd3g47550, Bd3g57890, Bd4g32020, Bd5g02730, Bd5g02760, Bd5g02770, Bd5g11740, respectively. And their gDNA and cDNA sequences were got by homology cloning.According to the bioinformatic analysis of amino that encoded by these genes, 12genes were predicted belonging to Pht1 gene family. The name were from BRAdi;Pht1;1- BRAdi;Pht1;12. The transmembrane structure analysis of which further defined them as Pht1, because they all had typical structure of Pht1 gene family. The results showed that all the predicted members have typical conserved structures of Pht1 gene and they are highly homological to each other, and they all had characteristic sequences GGDYPLSATIMSEYA.The cloned Pht1 members were divided into several subgroups by phylogenetic analysis, which revealed that Pht1 members from Brachypodium distachyon are highly homological to those from barley, but less homological to those from rice and Arabidopsis.Quantitative realtime PCRwas used for Tissue-specific expression analysis of Pht1members. Pht1 genes were differentially expressed in tissues.Most genes had the highest expression level in seeds. BdPht1;6,BdPht1;8,BdPht1;9 andBdPht1;12 had higher expression level in root samples than that in leaves of young seedlings.The material were treated with different concentration of phosphorus,and root RNAwere extracted for quantitative realtime PCR。The expression level of BdPht1;6(1g76010)were20 times of the check .The expression level decreased with the increase of concentration of phosphorus.Constructed Bd1g67330,Bd1g76010,Bd4g32020 into Arabidopsis vector PCAM1300and Yeast vector P11A1NE for Arabidopsis transgenic and yeast complementary experiments to verificated their function.The result still need further experiment.