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Plant Molecular Biology Reporter

, Volume 29, Issue 2, pp 265–277 | Cite as

Characterization of Upstream Sequences of the LOJ Gene Leads to Identification of a Novel Enhancer Element Conferring Lateral Organ Junction-Specific Expression in Arabidopsis thaliana

  • Dipnarayan Saha
  • Vajinder Kumar
  • Shripad Ramachandra Bhat
  • Ramamurthy Srinivasan
Article

Abstract

Isolation and characterization of promoters are important in understanding gene regulation and genetic engineering of crop plants. Earlier, a pentatricopeptide repeat protein (PPR) encoding gene (At2g39230), designated as Lateral Organ Junction (LOJ) gene, was identified through T-DNA promoter trapping in Arabidopsis thaliana. The upstream sequence of the LOJ gene conferred on the reporter gene a novel LOJ-specific expression. The present study was aimed at identifying and characterizing the cis-regulatory motifs responsible for tissue-specific expression in the −673 and +90 bases upstream of the LOJ gene recognized as LOJ promoter. In silico analysis of the LOJ promoter revealed the presence of a few relevant regulatory motifs and a unique feature like AT-rich inverted repeat. Deletion analysis of the LOJ promoter confirmed the presence of an enhancer-like element in the distal region (−673/−214), which stimulates a minimal promoter-like sequence in the −424/−214 region in a position and orientation autonomous manner. The −136/+90 region of the LOJ promoter was efficient in driving reporter gene expression in tissues like developing anthers and seeds of Arabidopsis. A positive regulation for the seed- and anther-specific expression module was contemplated within the 5′ untranslated region of the LOJ gene. However, this function was repressed in the native context by the lateral organ junction-specific expression. The present study has led to the identification of a novel lateral organ junction-specific element and an enhancer sequence in Arabidopsis with potential applications in plant genetic engineering.

Keywords

Arabidopsis Enhancer element Lateral organ junction promoter Pentatricopeptide repeat protein Promoter deletion analysis 

Abbreviations

CaMV

Cauliflower mosaic virus

DAG

Days after germination

GUS

β-Glucuronidase

LOJ

Lateral organ junction

MS

Murashige and Skoog

PCR

Polymerase chain reaction

PPR

Pentatricopeptide repeat protein

RT-PCR

Reverse transcription polymerase chain reaction

TSS

Transcription start site

UTR

Untranslated region

Notes

Acknowledgements

We thank Dr KV Prabhu (National Phytotron Facility, IARI) for his help in growing Arabidopsis in growth chambers and Greenhouse. We also thank Dr. S. Chandrasekharan of Genetics Division, IARI, in facilitating microscopic photographs. This research was funded in part by National Agricultural Technology Project (NATP), CGP grant (CGPII/253).

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Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Dipnarayan Saha
    • 1
  • Vajinder Kumar
    • 1
  • Shripad Ramachandra Bhat
    • 1
  • Ramamurthy Srinivasan
    • 1
  1. 1.National Research Center for Plant BiotechnologyIndian Agricultural Research InstituteNew DelhiIndia

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