Abstract
Δ1-pyrroline-5-carboxylate synthetase (P5CS) is one of the key regulatory enzymes involved in the proline biosynthetic pathway. Proline acts as an osmoprotectant, molecular chaperone, antioxidant, and regulator of redox homeostasis. The accumulation of proline during stress is believed to confer tolerance in plants. In this study, we cloned the complete CDS of the P5CS from pearl millet (Pennisetum glaucum (L.) R.Br. and transformed into tobacco. Three transgenic tobacco plants with single-copy insertion were analyzed for drought and heat stress tolerance. No difference was observed between transgenic and wild-type (WT) plants when both were grown in normal conditions. However, under heat and drought, transgenic plants have been found to have higher chlorophyll, relative water, and proline content, and lower malondialdehyde (MDA) levels than WT plants. The photosynthetic parameters (stomatal conductance, intracellular CO2 concentration, and transpiration rate) were also observed to be high in transgenic plants under abiotic stress conditions. qRT-PCR analysis revealed that the expression of the transgene in drought and heat conditions was 2–10 and 2–7.5 fold higher than in normal conditions, respectively. Surprisingly, only P5CS was increased under heat stress conditions, indicating the possibility of feedback inhibition. Our results demonstrate the positive role of PgP5CS in enhancing abiotic stress tolerance in tobacco, suggesting its possible use to increase abiotic stress-tolerance in crops for sustained yield under adverse climatic conditions.
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Acknowledgements
The authors are thankful to the Director NIPB, New Delhi, India for providing facilities to carry out this research. Director IARI (Indian Agricultural Research Institute), New Delhi, India is duly thanked for providing National Phytotron facilities. The support by ICAR (Indian Council of Agricultural Research) under the National Innovations in Climate Resilient Agriculture project till March 2020 is duly acknowledged. We thank RNDr. Cepl Jaroslav, FLD, CZU, for statistical support.
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GS carried out tobacco transformation, physiological and biochemical assays, molecular analysis, and manuscript writing. MC, AT isolated PgP5CS gene. RSJ carried out the bioinformatic analysis. AT, RSJ involved in manuscript editing. HV was involved in the physiological and 2D-SDS PAGE. JCP conceptualized the research plan, designed experiments, involved in manuscript preparation and editing. All the authors have read and approved the manuscript.
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Sellamuthu, G., Tarafdar, A., Jasrotia, R.S. et al. Introgression of Δ1-pyrroline-5-carboxylate synthetase (PgP5CS) confers enhanced resistance to abiotic stresses in transgenic tobacco. Transgenic Res (2024). https://doi.org/10.1007/s11248-024-00385-x
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DOI: https://doi.org/10.1007/s11248-024-00385-x