Functional characterization of two myo-inositol-1-phosphate synthase (MIPS) gene promoters from the halophytic wild rice (Porteresia coarctata)
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The promoter deletion mutants from second isoform of INO1 (gene-encoding MIPS) from Porteresia coarctata of 932 bp (pPcINO1.2.932) and 793 bp (pPcINO1.2.793) prove to be very efficient as salt/drought stress-inducible promoters, while pPcINO1.2.932 is found to be responsive to cold stress as well.
The promoters of the two identified myo-inositol-1-phosphate synthase (INO1) isoforms from salt-tolerant wild rice, Porteresia coarctata (PcINO1.1 and PcINO1.2) have been compared bioinformatically with their counterparts present in the salt-sensitive rice, Oryza sativa. PcINO1.2 promoter was found to be enriched with many abiotic stress-responsive elements, like abscisic acid-responsive elements, MYC-responsive elements, MYB-binding sites, low-temperature stress-responsive elements, and heat-shock elements similar to the ones found in the conserved motifs of the promoters of salt/drought stress-inducible INO1 promoters across Kingdom Planta. To have detailed analysis on the arrangement of cis-acting regulatory elements present in PcINO1 promoters, 5′ deletion mutational studies were performed in dicot model plants. Both transient as well as stable transformation methods were used to check the influence of PcINO1 promoter deletion mutants under salt and physiologically drought conditions using β-glucuronidase as the reporter gene. The deletion mutant from the promoter of PcINO1.2 of length 932 bp (pPcINO1.2.932) was found to be significantly upregulated under drought stress and also in cold stress, while another deletion mutant, pPcINO1.2.793 (of 793 bp), was significantly upregulated under salt stress. P. coarctata being a halophytic species, the high inducibility of pPcINO1.2.932 upon exposure to low-temperature stress was an unexpected result.
KeywordsPromoter deletion mutants Transient transformation Abiotic stress GUS reporter assay Post-transcriptional repression Stable transgenics
This work was supported by grants from the Department of Atomic Energy and Department of Biotechnology awarded to ALM, currently an INSA Senior Scientist; and PB was supported by Fellowships from the Council of Scientific and Industrial Research.
This study was funded by grants from the Department of Atomic Energy (D.O. No. 10/25/2010/RRF-R&D-II/3118) and Department of Biotechnology (BT/AB/05/02/2007-III) awarded to ALM, currently an INSA Senior Scientist. PB was supported by Fellowships from the Council of Scientific and Industrial Research [09/015(0447)/2012-EMR-I].
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Conflict of interest
The authors declare that they have no conflicts of interest.
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