The Douglas-fir BiP promoter is functional in Arabidopsis and responds to wounding.
A DNA sequence representing the promoter region of the Douglas-fir (Pseudotsuga menziesii [Mirb.] Franco) luminal binding protein PmBiP (PmBiPPro1) was isolated using inverse polymerase chain reaction (iPCR). Transient expression analysis of PmBiPPro1 fused to the beta-glucuronidase (GUS) reporter gene demonstrated that this promoter is functional in germinating Douglas-fir embryos. Transgenic Arabidopsis plants containing PmBiPPro1:GUS reporter gene constructs revealed strong staining associated with actively dividing/expanding cells and secretory tissues in developing seedlings. Wounding of cotyledons resulted in an increase in local staining associated with cells surrounding the wound site. Deletion analysis showed that elements necessary for basal-level expression reside within a -261 to +16 bp region, although upstream elements are necessary for higher-level expression in germinating Douglas-fir embryos, developing Arabidopsis seedlings and wounded cotyledons. Correlation of the observed expression pattern with the known function of BiP suggests that pathways controlling expression are highly conserved between angiosperms and gymnosperms.
Forward
Benjamin S
BS
Department of Biochemistry and Microbiology and Centre for Forest Biology, University of Victoria, P.O. Box 3055 STN CSC, Victoria, British Columbia V8W 3P6, Canada.
Osusky
Milan
M
Misra
Santosh
S
D017360:Arabidopsis / Q000235:genetics* / Q000502:physiology; D029681:Arabidopsis Proteins; D001483:Base Sequence; D002352:Carrier Proteins / Q000235:genetics* / Q000502:physiology; D003001:Cloning, Molecular; D018506:Gene Expression Regulation, Plant; D018525:Germination; D005966:Glucuronidase / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000235:genetics* / Q000502:physiology; D030821:Plants, Genetically Modified; D011401:Promoter Regions, Genetic / Q000235:genetics* / Q000502:physiology; D028224:Pseudotsuga / Q000235:genetics* / Q000502:physiology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012639:Seeds / Q000235:genetics / Q000254:growth & development; D013314:Stress, Mechanical
D029681:Arabidopsis Proteins; D002352:Carrier Proteins; D010940:Plant Proteins; D011993:Recombinant Fusion Proteins; C077221:BIP protein, Arabidopsis; D005966:Glucuronidase