Temporal and spatial control of transgene expression using a heat-inducible promoter in transgenic wheat.
Constitutive promoters are widely used to functionally characterise plant genes in transgenic plants, but their lack of specificity and poor control over protein expression can be a major disadvantage. On the other hand, promoters that provide precise regulation of temporal or spatial transgene expression facilitate such studies by targeting over-expression or knockdown of target genes to specific tissues and/or at particular developmental stages. Here, we used the uidA (beta-glucuronidase, GUS) reporter gene to demonstrate that the barley Hvhsp17 gene promoter can be induced by heat treatment of 38-40 °C for 1-2 h in transgenic wheat. The GUS enzyme was expressed only in those tissues directly exposed to heat and not in neighbouring leaf tissues. The induction of HSP::GUS was demonstrated in all organs and tissues tested, but expression in older tissues was lower. Generally, proximal root sections showed less GUS activity than in root tips. This heat-inducible promoter provides the ability to investigate the function of candidate genes by overexpression or by down-regulation of target gene expression (for example by RNAi) in selected tissues or developmental stages of a transgenic plant, limited only by the ability to apply a heat shock to the selected tissues. It also allows the investigation of genes that would be lethal or reduce fertility if expressed constitutively.
Rothamsted Research, Harpenden, Hertfordshire, UK. jackie.freeman@bbsrc.ac.uk
Jacqueline
J
Freeman
Caroline A
CA
Sparks
Jevon
J
West
Peter R
PR
Shewry
Huw D
HD
Jones
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D015536:Down-Regulation; D018506:Gene Expression Regulation, Plant / Q000235:genetics*; D017930:Genes, Reporter; D005966:Glucuronidase / Q000235:genetics / Q000378:metabolism; D006360:Heat-Shock Proteins / Q000235:genetics; D001467:Hordeum / Q000235:genetics; D006358:Hot Temperature; D009928:Organ Specificity; D018515:Plant Leaves / Q000166:cytology / Q000235:genetics / Q000502:physiology; D018517:Plant Roots / Q000166:cytology / Q000235:genetics / Q000502:physiology; D018547:Plant Stems / Q000166:cytology / Q000235:genetics / Q000502:physiology; D030821:Plants, Genetically Modified / Q000166:cytology / Q000235:genetics / Q000502:physiology*; D011058:Pollen / Q000166:cytology / Q000235:genetics / Q000502:physiology; D011401:Promoter Regions, Genetic / Q000235:genetics*; D018749:RNA, Plant / Q000235:genetics; D012639:Seeds / Q000166:cytology / Q000235:genetics / Q000502:physiology; D013997:Time Factors; D014018:Tissue Distribution; D019076:Transgenes; D014908:Triticum / Q000166:cytology / Q000235:genetics / Q000502:physiology*
D001426:Bacterial Proteins; D006360:Heat-Shock Proteins; D018749:RNA, Plant; D005966:Glucuronidase