Overexpression of PgDREB2A transcription factor enhances abiotic stress tolerance and activates downstream stress-responsive genes

Abstract

The DREB transcription factors comprise conserved ERF/AP2 DNA-binding domain, bind specifically to DRE/CRT motif and regulate abiotic stress mediated gene expression. In this study we show that PgDREB2A from Pennisetum glaucum is a powerful transcription factor to engineer multiple stress tolerance in tobacco plants. The PgDREB2A protein lacks any potential PEST sequence, which is known to act as a signal peptide for protein degradation. Therefore, the transgenic tobacco plants were raised using full-length cDNA without modification. The transgenics exhibited enhanced tolerance to both hyperionic and hyperosmotic stresses. At lower concentration of NaCl and mannitol, seed germination and seedling growth was similar in WT and transgenic, however at higher concentration germination in WT decreased significantly. D15 and D46 lines showed 4-fold higher germination percent at 200 mM NaCl. At 400 mM mannitol seed germination in WT was completely arrested, whereas in transgenic line it was more than 50%. Seedlings of D15 and D46 lines showed better growth like leaf area, root number, root length and fresh weight compared to wild type for both the stresses. The quantitative Real time PCR of transgenic showed higher expression of downstream genes NtERD10B, HSP70-3, Hsp18p, PLC3, AP2 domain TF, THT1, LTP1 and heat shock (NtHSF2) and pathogen-regulated (NtERF5) factors with different stress treatments.