5' deletion of a gbss1 promoter region from wheat leads to changes in tissue and developmental specificities.
Expression of granule-bound starch synthase 1 (GBSS1) in wheat is restricted to the grain filling process. In order to identify promoter regions which are involved in transcriptional control of the observed expression pattern, we isolated about 8 kb of a wheat gbss1-upstream region. Within this sequence several putative cis-acting elements were identified. In addition, an untranslated leader region is located in the 5' region of the gbss1 gene. To investigate promoter activity of the isolated region, the proximal 4.0 kb and progressively 5'-deleted fragments were transcriptionally fused to a beta-glucuronidase reporter gene. The function of the promoter constructs was tested by transient expression assays in various wheat tissues and in transgenic wheat plants, which were selected for low number and integrity of transgene copies. Analysis of stable transformants revealed that the -4.0 kb promoter region mediates reporter gene expression that is in accordance with the endogenous gbss1 expression. Promoter deletion to -1.9 kb or to -1.0 kb did not change the expression profile with regard to grain and pollen specificity. However, the profile of beta-glucuronidase expression during the grain filling process is altered in such a way that the level of beta-glucuronidase activity declines due to the decreasing promoter length. It is proposed that enhancer elements and cis-acting elements, which are involved in gbss1 transcription during the grain filling process, are located -1.9 kb upstream of the promoter. In addition, participation of the untranslated leader region in tissue-specific gene expression is discussed.
Kluth
Antje
A
Centre for Applied Plant Molecular Biology (AMP II), University of Hamburg, Germany.
Sprunck
Stefanie
S
Becker
Dirk
D
Lörz
Horst
H
Lütticke
Stephanie
S
D024506:5' Flanking Region / Q000235:genetics; D018744:DNA, Plant / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D005470:Fluorometry; D018506:Gene Expression Regulation, Plant; D005966:Glucuronidase / Q000235:genetics / Q000378:metabolism; D006651:Histocytochemistry; D030821:Plants, Genetically Modified; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D017422:Sequence Analysis, DNA; D017384:Sequence Deletion; D013214:Starch Synthase / Q000235:genetics*; D014158:Transcription, Genetic; D014908:Triticum / Q000235:genetics* / Q000254:growth & development
D018744:DNA, Plant; D011993:Recombinant Fusion Proteins; C434607:granule-bound starch synthase I; D013214:Starch Synthase; D005966:Glucuronidase