, Volume 248, Issue 5, pp 1121–1141 | Cite as

Functional characterization of two myo-inositol-1-phosphate synthase (MIPS) gene promoters from the halophytic wild rice (Porteresia coarctata)

  • Papri Basak
  • Shiny Sangma
  • Abhishek Mukherjee
  • Tanushree Agarwal
  • Sonali Sengupta
  • Sudipta Ray
  • Arun Lahiri MajumderEmail author
Original Article


Main conclusion

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.


Promoter 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].

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interest.

Supplementary material

425_2018_2957_MOESM1_ESM.pdf (72 kb)
Online Resource 1 (ESM_1.pdf): Primers designed for generation of deletion mutants for pPcINO1.1 and pPcINO1.2 (PDF 71 kb)
425_2018_2957_MOESM2_ESM.pdf (88 kb)
Online Resource 2 (ESM_2.pdf): Cis-acting regulatory elements (CAREs) found in INO1 promoters categorized according to their responsiveness towards abiotic, biotic stress and phytohormones (PDF 87 kb)
425_2018_2957_MOESM3_ESM.tif (3.4 mb)
Online Resource 3 (ESM_3.tif): Comparison of the stress-responsive cis-acting elements for pPcINO1.1, pPcINO1.2 and pOsINO1.1. Abbreviations used: DRE, LTRE (DRECRTCORE specifically responsive under Low-Temperature stress), MYB, DSRE (MYB Recognition Elements responsive to Dehydration Stress), HSE (Heat-Shock Element), P& SI (Pathogen and Salt-Inducible elements), ABRE (ABscisic acid-Responsive Elements), MYC (MYC-binding sites), ARE (Auxin-Responsive Elements), MSA (Meristem-Specific Activating elements), TATA box and CAAT box. CAAT boxes are represented in light grey font while TATA boxes are shown in italicized font (TIFF 3458 kb)
425_2018_2957_MOESM4_ESM.tif (1.6 mb)
Online Resource 4 (ESM_4.tif): Transient transformation experiments performed in Rice and Arabidopsis leaf discs using pPcINO1.2 deletion series. a GUS histochemical assay b Relative promoter activity calculated from quantitative analysis of GUS expression level (MUG assay) (TIFF 1688 kb)
425_2018_2957_MOESM5_ESM.tif (3.5 mb)
Online Resource 5 (ESM_5.tif): MALDI–TOF spectra obtained from the in-gel trypsin digested eluent fractions from affinity chromatography using rice nuclear proteins bound to repressor region (RR) a-d Spectra obtained from ~60 kDa band in 12%SDS from eluent fractions using 0.2 M (a), 0.4 M (b), 0.6 M (c) and 1 M KCl (d); e–g from ~40 kDa band for eluent fractions using 0.2 M (e), 0.4 M (f) and 0.6 M KCl (g); hand i from ~10 kDa band for eluent fractions using 0.2 M (h) and 0.4 M KCl (i) (TIFF 3583 kb)
425_2018_2957_MOESM6_ESM.pdf (118 kb)
Online Resource 6 (ESM_6.pdf): Mascot search results of the identified rice nuclear proteins in different eluent fractions with significant scores (i.e. >60) (PDF 117 kb)


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Division of Plant BiologyBose InstituteKolkataIndia
  2. 2.HSSLC BranchMeghalaya Board of School EducationShillongIndia
  3. 3.Department of BotanyUniversity of CalcuttaKolkataIndia
  4. 4.School of Plant, Environment and Soil SciencesLouisiana State University Agricultural CentreBaton RougeUSA

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