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

Abstract

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 T 1 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.

Variation in Rubisco activase (RCAβ) gene promoters and expression in soybean [Glycine max (L.) Merr.]

“…To identify whether sequence polymorphisms in the promoter region of GmRCAβ were related to the observed expression level, a 2300-bp region of the promoter, which was shown to be sufficient for the proper regulation of expression in plant (Orozco and Ogren, 1993; Liu et al , 1996; Qu et al , 2011; Yang et al , 2012), was cloned and sequenced, including the promoter and the 5′-UTR region, in a natural population including 219 landraces of different geographic origins (Supplementary Figs S4 and S5). Multiple sequence alignment revealed 16 polymorphic sites and five types of sequences (Fig.…”

“…In the present study, seven SNPs were identified as significantly associated with the expression of GmRCAβ (Table 5). Interestingly, three SNPs were located in regulatory elements such as the AT1 motif and the TATA box, which are important for the light regulation of gene expression, and data gathered in several species indicate that the expression of RCA is highly regulated by light (Qu et al , 2011). These SNPs in regulatory regions may impact the phenotype by altering the activity of the promoter and changing the expression levels of the RCA genes (Kumar et al , 2010).…”

“…In contrast to the RCA gene and its encoded protein, which have been well studied in a number of species, the molecular mechanism of transcriptional regulation about RCA gene has thus far been only studied in spinach (Orozco and Ogren, 1993), Arabidopsis (Liu et al , 1996), potato (Qu et al , 2011), and rice (Yang et al , 2012). Gene transcription is a primary intermediate between the information encoded in the genome and the final phenotype.…”

rbcS SRS4 promoter from Glycine max and its expression activity in transgenic tobacco

“…4) GATA motifs (WGATAR); these are implicated to act in the light-dependent and nitrate-dependent control of transcription in plants (Reyes et al, 2004). 5) Motifs with a known relationship to light including H box (CCTACC), REalpha element (AACCAA), and CCAAT-box (Song et al, 2000;Outchkourov et al, 2003;Qu et al, 2011). The cis-acting elements identified in rbcSP strongly suggested that heterologous genes driven by rbcSP could be induced in a lightspecific manner.…”

“…The rbcS promoter has been cloned, sequenced, and studied in various plants, including pea (Pisum sativum; Nagy et al, 1985), cotton (Gossypium spp; Song et al, 2000), Chrysanthemum (Outchkourov et al, 2003), coffee (Coffea arabica; Marraccini et al, 2003), and potato (Solanum tuberosum L.; Qu et al, 2011). In soybean, the rbcS gene family comprises 6-10 members (Dean et al, 1989).…”