The Study And Modification Of Amino Acid Property Of Arylpyrazole Class Guided Pesticides

Guided pesticides are utilizing the certain active transporter to reach the optimal distribution in plant by adding endogenous amino acid or hexose derivatives to the structure of pesticides.In order to improve this theory,further study was accoplised in Ricinus Communis,Arabidopsis thaliana,and Xenopus Oocyte system,based on the study on GlyF and GlyMeF.The aim of this syudy is to investigate the amino acid property of arylpyrazole alass guided pesticides in molecular biology perspective.At the same time,the modification of those compound has also been finshed.Thus,could help the further study of guided pesticides.Cotyledon of R.communis seedling were teratd by 0.1 mM GlyMeF.Then the expression profile of RcLHT6,RcANT15,RcProT2,RcCAT2 has been shown by RT-qPCR test.RcANT15 was not upregulation.Thus,this gene wes selected to build the methold of X.Oocyte drug uptake experiment.Oocyte with different volume of RcANT15 cRNA injection were selected to perform time course experiment of 0.1 mM GlyF uptake.The result shows the best injact volume was 18.4nl and proper experiment time was 1h.Oocyte with RcLHT6,RcANT15,RcProT2,and RcCAT2 cRNA injection,were chosen to uptake 0.1mM GlyF or GlyMeF.The injection of genes did not help the uptake of GlyMeF.However,RcANT15 increased almost 3.5 times of GlyF uptake,compared with control,the rest genes did not show significant impact.This result indicates that RcANT15 had the fuction of GlyF uptake.After the modification of GlyF structure by simplify the structure,and netcharge adjusttion with the help of ACD LogD,GlnF was designed.GlnF uptake cotyledon of R.communis seedling indicated the saturable profile at low concentration.The uptake is sensitive to pH,CCCP,and amino acid,which fit the profile of amino acid uptake.The phloem loading test also indicatd the involvement of amino acid carrier in the phloem loading process.The root of Wild type,AtLHT1 knock out or overexpression A.thaliana were used to perform root uptake experiments of 0.05 mM GlnF.The uptake of GlnF could not detecte in the AtLHT1 knock out plant.The overexpress plant has 75% more uptake compare to wild type.This has proved AtLHT1 could transport GlnF in planta level.Meanwhile,based on the study on Arabidopsis thaliana the process of uptaking exogenous amino acid is responsible for GlnF uptake.The result of oocyte experiment towards 0.05 mM GlnF and AtLHT1 is consistant with those in Arabidopsis thaliana experiment.