Research Article

Isolation and characterization of the organ-specific and light-inducible promoter of the gene encoding rubisco activase in potato (Solanum tuberosum)

Published: April 12, 2011
Genet. Mol. Res. 10 (2) : 621-631 DOI: https://doi.org/10.4238/vol10-2gmr1088
Cite this Article:
(2011). Isolation and characterization of the organ-specific and light-inducible promoter of the gene encoding rubisco activase in potato (Solanum tuberosum). Genet. Mol. Res. 10(2): gmr1088. https://doi.org/10.4238/vol10-2gmr1088
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Abstract

Constitutive promoters have been widely used in crop biotechnology applications. Tissue-specific or inducible promoters, however, have advantages in some cases. We isolated the 731-bp 5' flanking sequence of a potato (Solanum tuberosum) gene, encoding ribulose-1,5-bisphosphate carboxylase/oxygenase (rubisco) activase (RCA), which was isolated by genome walking. By using GUS as a reporter and with Northern blot analysis, the 702-bp fragment (referred to as StRCAp), ranging from nt -731 to -30 relative to the initiation code of the RCA gene, was analyzed in transgenic tobacco plants. The activity of StRCAp in leaves was 0.4-fold less than that of cauliflower mosaic virus 35S promoter, and was expressed throughout the green part of the light-grown transgenic T1 seedlings, including cytoledons, leaves and young stems, but not roots. Further deletion analysis revealed that a shorter fragment (nt -249 to -30, StRCAp2) retained light-inducible features in cytoledons and leaves, but showed no detectable activity in young stems and roots. Although the activity of StRCAp2 in leaves was reduced significantly compared with that of StRCAp, the overall data indicated that cis-elements sufficient to regulate organ-specific and light-inducible transcription are within the 220-bp fragment. There is potential for application of StRCAp in plant genetic engineering.

Constitutive promoters have been widely used in crop biotechnology applications. Tissue-specific or inducible promoters, however, have advantages in some cases. We isolated the 731-bp 5' flanking sequence of a potato (Solanum tuberosum) gene, encoding ribulose-1,5-bisphosphate carboxylase/oxygenase (rubisco) activase (RCA), which was isolated by genome walking. By using GUS as a reporter and with Northern blot analysis, the 702-bp fragment (referred to as StRCAp), ranging from nt -731 to -30 relative to the initiation code of the RCA gene, was analyzed in transgenic tobacco plants. The activity of StRCAp in leaves was 0.4-fold less than that of cauliflower mosaic virus 35S promoter, and was expressed throughout the green part of the light-grown transgenic T1 seedlings, including cytoledons, leaves and young stems, but not roots. Further deletion analysis revealed that a shorter fragment (nt -249 to -30, StRCAp2) retained light-inducible features in cytoledons and leaves, but showed no detectable activity in young stems and roots. Although the activity of StRCAp2 in leaves was reduced significantly compared with that of StRCAp, the overall data indicated that cis-elements sufficient to regulate organ-specific and light-inducible transcription are within the 220-bp fragment. There is potential for application of StRCAp in plant genetic engineering.