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http://nopr.niscpr.res.in/handle/123456789/29138| Title: | Inducibility of dehydration responsive element (DRE)-based promoter through gusA expression in transgenic tobacco |
| Authors: | Basu, Supratim Roychoudhury, Aryadeep |
| Keywords: | Cold stress;Dehydration responsive element (DRE);β-glucuronidase (GUS);Polyethylene glycol;Promoter inducibility;Salinity;Transgenic tobacco;Water deficit |
| Issue Date: | Apr-2014 |
| Publisher: | NISCAIR-CSIR, India |
| Abstract: | The current study focuses on the pattern of stress-inducibility of a synthetically designed promoter, viz., 4X DRE (four tandem repeats of dehydration-responsive element), in response to the various stress-inducers like NaCl-mediated salinity, polyethylene glycol (PEG)-mediated water deficit, cold and abscisic acid (ABA). The construct, containing the 4X DRE promoter linked to the reporter gene gusA (that encodes β-glucuronidase, GUS), was introduced in tobacco plants via Agrobacterium tumefaciens-mediated transformation. The T2 progenies showed the integration of gusA, as verified by polymerase chain reaction (PCR) and DNA Blot hybridization analysis. The gusA induction was noted upon treatment of T2 transgenics with 20% (w/v) PEG for 24 h, while remained undetected even in PEG-treated wild type (WT) plant. The maximum expression was observed in the transgenic plant T2/3, which also showed induction in gusA expression with other stressors, viz., 200 mM NaCl and cold (4°C). However, PEG-mediated water stress seemed to be the most effective signal for promoter activation, followed by salinity stress and with lesser stimulation by cold. The promoter was activated in a time-dependent manner of stress application, i.e., greater gusA expression was detected after 24 h of stress as compared to 6 h. ABA application, even at a concentration of 100 μM for 24 h, failed to activate reporter gene expression, proving that the promoter is stress-inducible but ABA-independent. Our observation showed the potentiality of 4X DRE for use as a stress-inducible promoter in overexpressing transgene(s) for salinity, drought and cold tolerance. |
| Page(s): | 172-177 |
| ISSN: | 0975-0967 (Online); 0972-5849 (Print) |
| Appears in Collections: | IJBT Vol.13(2) [April 2014] |
Files in This Item:
| File | Description | Size | Format | |
|---|---|---|---|---|
| IJBT 13(2) 172-177.pdf | 377.62 kB | Adobe PDF | View/Open |
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