Testing of a heterologous, wound- and insect-inducible promoter for functional genomics studies in conifer defense.
Large-scale sequencing of conifer cDNAs and targeted molecular cloning have identified many putative conifer defense genes. Expression of many of these genes is induced in response to biotic stress and some may be expressed only in a few specialized tissues or cells. Proven functional genomics approaches to test these genes involve expression of proteins in Escherichia coli or yeast for biochemical characterization or constitutive over-expression in transformed plants. Plant transformation to test functions of insect-, wound- or pathogen-induced conifer defense genes would benefit from the use of an inducible expression system. We describe here the development of a heterologous, wound- and insect-inducible gene expression system for conifers using the potato proteinase inhibitor II (pinII)-promoter. Using pinII::GUS and pinII::(E)-alpha-bisabolene synthase expression constructs we demonstrate localized induced gene expression in white spruce seedlings (Picea glauca). Testing of these constructs in Arabidopsis thaliana and tobacco illustrates the additional potential of the pinII-promoter to be used in tests of gene function that involve cell-specific and systemic induction.
Michael Smith Laboratories, University of British Columbia, Vancouver, BC, Canada.
Kimberley-Ann
KA
Godard
Ashley
A
Byun-McKay
Caroline
C
Levasseur
Aine
A
Plant
Armand
A
Séguin
Jörg
J
Bohlmann
D000818:Animals; D017360:Arabidopsis / Q000235:genetics / Q000254:growth & development / Q000469:parasitology; D018506:Gene Expression Regulation, Plant; D018745:Genome, Plant; D023281:Genomics / Q000379:methods*; D007113:Immunity, Innate / Q000235:genetics; D007313:Insecta / Q000254:growth & development*; D028222:Picea / Q000235:genetics / Q000254:growth & development / Q000469:parasitology; D010935:Plant Diseases / Q000235:genetics / Q000469:parasitology; D010940:Plant Proteins / Q000235:genetics / Q000502:physiology; D030821:Plants, Genetically Modified; D011401:Promoter Regions, Genetic / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013314:Stress, Mechanical; D014026:Nicotiana / Q000235:genetics / Q000254:growth & development / Q000469:parasitology; D064028:Tracheophyta / Q000235:genetics* / Q000254:growth & development / Q000469:parasitology
D010940:Plant Proteins