Tissue-specific activation of the osmotin gene by ABA, C2H4 and NaCl involves the same promoter region.
The gene encoding the antifungal protein osmotin is induced by several hormonal and environmental signals. In this study, tissue-specific and inducer-mediated expression of the reporter gene beta-glucuronidase (uidA) fused to different fragment lengths of the osmotin promoter was evaluated in transgenic tobacco (Nicotiana tabacum). The region of the promoter between -248 to -108 (Fragment A) was found to be essential and sufficient for inducer (abscisic acid (ABA), C2H4 and NaCl)-mediated expression of the reporter gene. Expression of the reporter gene was developmentally regulated and increased with maturity of leaves, stem and flowers. Expression also was tissue-specific being most highly expressed in epidermis and vascular parenchyma of the stem. The regulators ABA, C2H4 and NaCl exhibited tissue-specific induction of this promoter. The promoter was specifically responsive to C2H4 in flowers at virtually all stages of development, but not responsive in these tissues to ABA or NaCl. Conversely, ABA and NaCl were able to induce reporter gene activity using promoter Fragment A in specific tissues of root where C2H4 was unable to induce activity. Further dissection of the promoter Fragment A into fragments containing either the conserved GCC element (PR); PR/AT; or G/AT sequences, and subsequent testing of these fragments fused to GUS in transgenic plants was performed. These experiments revealed that the promoter fragment containing PR element alone, although required, was barely able to allow responsiveness to C2H4. However, significant C2H4-induced activity was obtained with a promoter fragment containing the AT and PR elements together.
Raghothama
K G
KG
Center for Plant Environmental Stress Physiology, Purdue University, West Lafayette, IN 47907-1165, USA.
Maggio
A
A
Narasimhan
M L
ML
Kononowicz
A K
AK
Wang
G
G
D'Urzo
M P
MP
Hasegawa
P M
PM
Bressan
R A
RA
D000040:Abscisic Acid / Q000494:pharmacology*; D001483:Base Sequence; D017124:Conserved Sequence; D018548:Cotyledon; D005030:Ethylenes / Q000494:pharmacology*; D018506:Gene Expression Regulation, Plant / Q000187:drug effects*; D017930:Genes, Reporter; D005966:Glucuronidase / Q000096:biosynthesis; D018515:Plant Leaves; D010940:Plant Proteins / Q000096:biosynthesis*; D018517:Plant Roots; D030821:Plants, Genetically Modified; D010947:Plants, Toxic; D011401:Promoter Regions, Genetic / Q000187:drug effects; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D017384:Sequence Deletion; D012965:Sodium Chloride / Q000494:pharmacology*; D014026:Nicotiana
D005030:Ethylenes; D010940:Plant Proteins; D011993:Recombinant Fusion Proteins; C073139:osmotin protein, Nicotiana tabacum; D012965:Sodium Chloride; D000040:Abscisic Acid; C036216:ethylene; D005966:Glucuronidase