Isolation and characterization of the 4coumarate

Authors: Huynh Thi Thu Hue Duong Thi Thu Ha Nong Van Hai Le Thi Thu Hien

Publish Date: 2016/08/11

Volume: 22, Issue: 3, Pages: 399-405

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Abstract

The most important enzyme of the phenylpropanoid pathway 4coumaratecoenzyme A ligase 4CL is encoded by several homologous genes including 4CL1 The 4CL1 promoter is a tissuespecific gene expression element particularly active in the secondary xylem or older stems In this study the 1127 bp 5′ upstream region of the 4CL1 coding sequence from Eucalyptus camaldulensis Euc4CL1 was isolated and characterized Essential putative ciselements in the Euc4CL1 promoter included a TATAbox at −22/−28 position two CCAATboxes at −256/−260 and −277/−281 positions respectively an ACelement at −328/−336 and Aboxes at −115/−120 and −990/−995 positions To investigate the effect of the Euc4CL1 promoter on gene expression a plant transformation vector pEuc4CL1p containing the reporter gene for βglucuronidase GUS under the control of Euc4CL1 promoter was constructed based on the pBI101 backbone and introduced in tobacco plants Stable expression of the GUS gene in transgenic lines was analysed by a histochemical GUS assay The results indicated the specific expression of the GUS gene in the stem xylem cells of transgenic tobacco lines was controlled by the Euc4CL1 promoter The observations suggest the isolated Euc4CL1 promoter is a potential candidate for driving the expression of a foreign gene in plant xylem tissuesWe would like to thank Prof Dr Alan B Bennett University of California Davis Dr Cecilia L ChiHam HM Clause for critical review of the manuscript We are also grateful to two anonymous reviewers for their helpful comments and criticisms This research was supported by the Research Grant No CNSHDT03/0610 from the Vietnam Ministry of Agriculture and Rural Development

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