Characterization of the ethanol-inducible alc gene-expression system in Arabidopsis thaliana.
Controlled expression of transgenes in plants is key to the characterization of gene function and the regulated manipulation of growth and development. The alc gene-expression system, derived from the filamentous fungus Aspergillus nidulans, has previously been used successfully in both tobacco and potato, and has potential for use in agriculture. Its value to fundamental research is largely dependent on its utility in Arabidopsis thaliana. We have undertaken a detailed function analysis of the alc regulon in A. thaliana. By linking the alcA promoter to beta-glucuronidase (GUS), luciferase (LUC) and green fluorescent protein (GFP) genes, we demonstrate that alcR-mediated expression occurs throughout the plant in a highly responsive manner. Induction occurs within one hour and is dose-dependent, with negligible activity in the absence of the exogenous inducer for soil-grown plants. Direct application of ethanol or exposure of whole plants to ethanol vapour are equally effective means of induction. Maximal expression using soil-grown plants occurred after 5 days of induction. In the majority of transgenics, expression is tightly regulated and reversible. We describe optimal strategies for utilizing the alc system in A. thaliana.
Roslan
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HA
Donnan Laboratories and Life Sciences Building, School of Biological Sciences, University of Liverpool, L69 7ZD, UK.
Salter
M G
MG
Wood
C D
CD
White
M R
MR
Croft
K P
KP
Robson
F
F
Coupland
G
G
Doonan
J
J
Laufs
P
P
Tomsett
A B
AB
Caddick
M X
MX
D017360:Arabidopsis / Q000235:genetics*; D001233:Aspergillus nidulans / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics*; D004790:Enzyme Induction; D000431:Ethanol / Q000494:pharmacology*; D005656:Fungal Proteins / Q000235:genetics*; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D018506:Gene Expression Regulation, Plant / Q000187:drug effects*; D017930:Genes, Reporter; D005966:Glucuronidase / Q000096:biosynthesis / Q000235:genetics; D049452:Green Fluorescent Proteins; D007700:Kinetics; D008156:Luciferases / Q000096:biosynthesis / Q000235:genetics; D008164:Luminescent Proteins / Q000235:genetics; D030821:Plants, Genetically Modified; D011401:Promoter Regions, Genetic; D018085:Regulon; D014026:Nicotiana / Q000235:genetics; D014170:Transformation, Genetic
C074202:ALCR protein, Aspergillus nidulans; D004268:DNA-Binding Proteins; D005656:Fungal Proteins; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D000431:Ethanol; D008156:Luciferases; D005966:Glucuronidase