Cloning And Functional Analysis Of ACO1Promoter In Vitis Vinifera

Vitis vinifera,which is one of the main raw materials of producing wine and has higheconomic value,contains high content of sugar and tastes sweet.While the resistance of Vitisvinifera is poor like the diseases caused by insects and environmental changes in its wholelife,which will affect its quality and yield seriously.Studies have shown that ethylene plays animportant roles in plant resistances,and ACC oxidase is the key enzyme in Ethylenebiosynthesis,which can catalytic ACC(1-aminocyclopropane-1-carboxylic acid)to ethylenedirectly.The it has important significance to study the molecular mechanisms of expressionand regulation of ACC oxidase.In this paper,the cloning and functional analysis of ACO1gene promoter in Vitis vinifera, which provide a theoretical basis for grape resistance study.The VvACO1gene promoter sequence is cloned from Vitis vinifera Genom DNA bygeneral PCR technology,which is1956bp length.This gene promoter sequence contains manyplant stress reponse elements(antioxidant response element,thermal shock stress element)andplant hormone response elements (abscisic acid-,auxin-,gibberellins-and salicylicacid-response element) obtained from cis acting element analysis of wed site PlantCARE.According to the analysis results of cis acting element in VvACO1gene promoter, thispaper amplificated5’-deletion fragment and acquired four fragments which are differentlengths(VvACO1P1~VvACO1P4),The four fragments are used to replace35S promoter ofPBI221-LUC,which finishes construction of recombinant expression vector.Transfer recombinant expression vectors to Arabidopsis thaliana protoplasts byPEG-Ca2+and test the expression leve of LUC reporter gene, this study finds that P1-P4fragments in VvACO1gene promoter can regulation the expression of downstream LUCreporter gene and-1483~-515bp sequence inhibites the expression of downstream gene and-515~-88bp sequence promotes.Allover,this study determine the species,number and characteristics of cis actingelements in VvACO1gene promoter,which provide experimental data for studing theexpression and regulation mechanisms and theory reference for the study of improving graperesistance.