Promoter of the wheat lipid transfer protein, TdLTP4, drives leaf-preferential expression in transgenic Arabidopsis plants

Abstract

In a previous report, a gene encoding a durum wheat lipid transfer protein, TdLTP4, was characterised as induced by abiotic and biotic stresses. In the present work, we investigated the regulation of the gene TdLTP4. A TdLTP4 promoter (PrTdLTP4) region of around 868-bp was isolated and sequenced. Its analysis revealed the presence of several DNA boxes known to be important mainly in the regulation of genes expressed under abiotic stress (salt and dehydration), abscisic acid (ABA) and pathogen responsiveness. The whole PrTdLTP4 fragment was fused to the reporter gene β-glucuronidase (gusA) and analysed in transgenic Arabidopsis plants. Histochemical assays of transgenic Arabidopsis plants showed that the 868-bp fragment of TdLTP4 gene promoter was found to be sufficient for both spatial and temporal patterns of its expression. Under control conditions, GUS histochemical staining was observed significantly only in young leaves of 8- and 12-day-old plants. Whereas after stress challenge especially with NaCl and mannitol, GUS transcripts expression increased substantially in leaves of 30-day-old transgenic seedlings. Real-time qPCR expression analysis of the gusA gene, confirmed the results of histochemical assays. Taken together these data provide evidence that PrTdLTP4 functions as abiotic-stress-inducible promoter in a heterologous dicot system and could be an excellent tool for future crop improvement.