Characterization of a strong, constitutive mung bean (Vigna radiata L.) promoter with a complex mode of regulation in planta

Cazzonelli, Christopher I., McCallum, Emily J., Lee, Rebecca and Botella, Jose Ramon (2005) Characterization of a strong, constitutive mung bean (Vigna radiata L.) promoter with a complex mode of regulation in planta. Transgenic Research, 14 6: 941-967. doi:10.1007/s11248-005-2539-2


Author Cazzonelli, Christopher I.
McCallum, Emily J.
Lee, Rebecca
Botella, Jose Ramon
Title Characterization of a strong, constitutive mung bean (Vigna radiata L.) promoter with a complex mode of regulation in planta
Journal name Transgenic Research   Check publisher's open access policy
ISSN 0962-8819
Publication date 2005-12-01
Year available 2005
Sub-type Article (original research)
DOI 10.1007/s11248-005-2539-2
Open Access Status Not yet assessed
Volume 14
Issue 6
Start page 941
End page 967
Total pages 27
Editor Bruce Whitelaw
Paul Christou
Place of publication Netherlands
Publisher Kluwer Academic
Language eng
Subject C1
270201 Gene Expression
620100 Field Crops
Abstract We report the cloning and characterization in tobacco and Arabidopsis of a Vigna radiata L. (mung bean) promoter that controls the expression of VR-ACS1, an auxin-inducible ACC synthase gene. The VR-ACS1 promoter exhibits a very unusual behavior when studied in plants different from its original host, mung bean. GUS and luciferase in situ assays of transgenic plants containing VR-ACS1 promoter fusions show strong constitutive reporter gene expression throughout tobacco and Arabidopsis development. In vitro quantitative analyses show that transgenic plants harboring VR-ACS1 promoter-reporter constructs have on average 4-6 fold higher protein and activity levels of both reporter genes than plants transformed with comparable CaMV 35S promoter fusions. Similar transcript levels are present in VR-ACS1 and CaMV 35S promoter lines, suggesting that the high levels of gene product observed for the VR-ACS1 promoter are the combined result of transcriptional and translational activation. All tested deletion constructs retaining the core promoter region can drive strong constitutive promoter activity in transgenic plants. This is in contrast to mung bean, where expression of the native VR-ACS1 gene is almost undetectable in plants grown under normal conditions, but is rapidly and highly induced by a variety of stimuli. The constitutive behavior of the VR-ACS1 promoter in heterologous hosts is surprising, suggesting that the control mechanisms active in mung bean are impaired in tobacco and Arabidopsis. The 'aberrant' behavior of the VR-ACS1 promoter is further emphasized by its failure to respond to auxin and cycloheximide in heterologous hosts. VR-ACS1 promoter regulatory mechanisms seem to be different from all previously characterized auxin-inducible promoters.
Keyword Biochemical Research Methods
Biochemistry & Molecular Biology
Biotechnology & Applied Microbiology
Acc Synthase
Cauliflower Mosaic Virus 35s
Constitutive Expression
Mung Bean
Plant Promoters
Stable Expression
1-aminocyclopropane-1-carboxylate Synthase Gene
Cauliflower-mosaic-virus
Auxin Response Elements
Transcript Flt Promoter
Cis-acting Element
Arabidopsis-thaliana
Transgenic Plants
Escherichia-coli
Aux/iaa Proteins
Messenger-rna
Q-Index Code C1
Institutional Status UQ
Additional Notes DOI: 10.1007/s11248-005-2539-2

 
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Created: Wed, 15 Aug 2007, 17:14:20 EST