, Volume 231, Issue 5, pp 1211–1227 | Cite as

Identification and organization of chloroplastic and cytosolic l-myo-inositol 1-phosphate synthase coding gene(s) in Oryza sativa: comparison with the wild halophytic rice, Porteresia coarctata

  • Sudipta Ray
  • Barunava Patra
  • Aparajita Das-Chatterjee
  • Arnab Ganguli
  • Arun Lahiri Majumder
Original Article


The gene coding for rice chloroplastic l-myo-inositol-1-phosphate synthase (MIPS; EC has been identified by matrix-assisted laser desorption time-of-flight mass spectrometry analysis of the purified and immunologically cross-reactive ~60 kDa chloroplastic protein following two-dimensional polyacrylamide gel electrophoresis, which exhibited sequence identity with the cytosolic MIPS coded by OsINO1-1 gene. A possible chloroplastic transit peptide sequence was identified upstream of the OsINO1-1 gene upon analysis of rice genome. RT-PCR and confocal microscope studies confirmed transcription, effective translation and its functioning as a chloroplast transit peptide. Bioinformatic analysis mapped the chloroplastic MIPS (OsINO1-1) gene on chromosome 3, and a second MIPS gene (OsINO1-2) on chromosome 10 which lacks conventional chloroplast transit peptide sequence as in OsINO1-1. Two new PcINO1 genes, with characteristic promoter activity and upstream cis-elements were identified and cloned, but whether these proteins can be translocated to the chloroplast or not is yet to be ascertained. Electrophoretic mobility shift assay carried out with nuclear extract of Porteresia coarctata leaves grown under both control and stressed condition shows binding of nuclear proteins with the upstream elements. Nucleotide divergence among the different Oryza and Porteresia INO1 genes were calculated and compared.


Chloroplastic l-myo-inositol-1-phosphate synthase MALDI-TOF MS Transit peptide GFP fluorescence Genomewalking Porteresia coarctata 



l-myo-Inositol-1-phosphate synthase


Matrix-assisted laser desorption ionization time-of-flight mass spectrometry


Cauliflower mosaic virus


Green fluorescent protein


Two-dimensional polyacrylamide gel electrophoresis


Open reading frame





The work is supported by grants to A.L.M. from the Department of Biotechnology, Government of India. S.R. and A.D.C. thank the Council of Scientific and Industrial Research, Government of India for Research Fellowships. Thanks are due to Dr Anka Hanemanne and Prof Klaus Eschrich, Institute of Biochemistry, Faculty of Medicine, University of Leipzig, Liebigstrasse 16, Leipzig D-04103, Germany, for help in analysis of the MALDI-TOF data.


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Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Sudipta Ray
    • 1
    • 2
  • Barunava Patra
    • 1
    • 3
  • Aparajita Das-Chatterjee
    • 1
    • 4
  • Arnab Ganguli
    • 1
    • 5
  • Arun Lahiri Majumder
    • 1
  1. 1.Plant Molecular and Cellular GeneticsBose Institute (Centenary Campus)KolkataIndia
  2. 2.Department of Botany, Centre of Advanced StudiesUniversity of CalcuttaKolkataIndia
  3. 3.Kentucky Tobacco Research and Development CentreUniversity of KentuckyLexingtonUSA
  4. 4.Molecular and Cell Biology Department, Goldman School of Dental MedicineBoston University Medical CenterBostonUSA
  5. 5.ChembiotekKolkataIndia

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