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Differential regulation of small heat-shock genes in plants: analysis of a water-stress-inducible and developmentally activated sunflower promoter

AuthorsCoca, María ; Almoguera, Concepción ; Thomas, T.L.; Jordano, Juan
Small heat-shock protein promoters
Differential regulation
Zygotic embryogenesis
Water stress
Abscisic acid response
Transgenic tobacco
Protein genes
Issue Date13-May-1996
PublisherKluwer Academic Publishers
CitationPlant Molecular Biology 31(4): 863-876, (1996).
AbstractWe have isolated two sunflower genes, Ha hsp 18.6 G2 and Ha hsp 17.7 G4, that encode small heat shock proteins (sHSPs). RNAse A protection experiments, carried out with RNA probes transcribed from each gene and hybridized to sunflower total RNA, allowed us to distinguish their mRNA accumulation patterns. In sunflower, Ha hspl7.7 G4 mRNAs accumulated during zygotic embryogenesis at 25 °C. In vegetative tissues, these mRNAs accumulated in response to either heat shock (42 °C), abscisic acid (ABA), or mild water stress treatments. In all cases, the mRNAs were transcribed from the same initiation site. In contrast, Ha hsp 18.6 G2 mRNAs accumulated only in response to heat-shock. This result demonstrates differential regulation of these two sHSP genes. The complex regulation depicted by the Ha hsp 17.7 G4 promoter has been further analyzed in transgenic tobacco, using G4::GUS translational fusions. Developmental induction of Ha hsp 17.7 G4 during zygotic embryogenesis was faithfully reproduced in the transgenic plants. 5'-distal sequences (between -1132 and -395) were required to confer a preferential spatial expression of GUS activity in the cotyledons. More proximal sequences (from -83 to +163) conferred to the chimeric genes most of the developmental regulation, and the responses to ABA and heat shock characteristic of the Ha hspl7.7 G4 promoter. The water stress response of this gene was not reproduced in transgenic tobacco and, thus, could be uncoupled from its regulation during embryogenesis.
DescriptionThe nucleotide sequence data reported will appear in the EMBL, GenBank and DDBJ Nucleotide Sequence Databases under the accession numbers U46544 (Ha hsplS.6 G2) and U46545 (Ha hsp 17.7 G4).
Publisher version (URL)http://www.springerlink.com/content/w1714v607022687g/fulltext.pdf
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