RESUMO
Pearl millet (Pennisetum glaucum) is an important cereal of traditional farming systems that has the natural ability to withstand various abiotic stresses. The present study aims at the identification and validation of major differentially expressed genes in response to drought stress in P. glaucum by Suppression Subtractive Hybridization (SSH) analysis. Twenty-two days old seedlings of P. glaucum cultivar PPMI741 were subjected to drought stress by treatment of 30 % Polyethylene glycol for different time periods 30 min (T1), 2 h (T2), 4 h (T3), 8 h (T4), 16 h (T5), 24 h (T6) and 48 h (T7) respectively, monitored by examining the RWC of seedlings. Total RNA was isolated to construct drought responsive subtractive cDNA library through SSH, sequenced to identify the differentially expressed genes in response to drought stress and validated by qRT-PCR.745 ESTs were assembled into a collection of 299 unigenes having 52 contigs and 247 singletons. All 745 ESTs were submitted to ENA-EMBL databases (Accession no. HG516611- HG517355). After analysis, 10 differentially expressed genes were validated namely Abscisic stress ripening protein, Ascorbate peroxidase, Inosine-5'-monophosphate dehydrogenase, Putative beta-1, 3-glucanase, Glyoxalase, Rab7, Aspartic proteinase Oryzasin, DnaJ-like protein and Calmodulin-like protein by qRT-PCR. The identified ESTs reveal a major portion of the stress responsive transcriptome that may prove to be a vent to unravel molecular basis underlying tolerance of pearl millet (Pennisetum glaucum) to drought stress. These genes could be utilized for transgenic breeding or transferred to crop plants through marker assisted selection for the development of better drought resistant cultivars having enhanced adaptability to survive harsh environmental conditions.