Promoter characterization of chickpea delta-1-pyrroline-5-carboxylate synthetase (P5CS) gene provides novel insights into its stress responsiveness
Main Article Content
Abstract
Proline is an osmolyte that accumulates in response to various environmental stresses and serves several protective functions in plants. The gene P5CS (delta-1-Pyrroline-5-carboxylate synthetase) codes for a key regulatory enzyme in proline biosynthesis. In the present study, we have isolated and in silico characterized the promoter region of the chickpea P5CS gene (CaP5CS). The expression of the gene was examined under various abiotic stresses such as cold, salinity and dehydration and also on application of various phytohormones and chemicals to understand the changes in gene expression which are driven by the promoter. Structurally, the promoter sequence was enriched in many cis- regulatory elements (CREs) recognized by transcription factors (TFs) involved in both ABA (Abscisic acid)-dependent and independent signaling pathways for proline biosynthesis. The gene was observed to be both spatially and temporally regulated. It was observed that the gene was highly up-regulated under heat and dehydration stress at 3 hours of stress treatment. Under dehydration, for the same tissues, the proline content was also estimated to increase by more than 3 fold from 3 to 6 hrs. Notably, under cold and IAA (indole-3-acetic acid) treatment, the gene was down-regulated, which confirms the role of the gene primarily under osmotic stress. This study provides novel insights into regulation of proline biosynthesis in chickpea. Also, the promoter isolated can be utilized to enable spatial and temporal control in transgene expression in genetically modified crops developed for enhanced stress tolerance.
Downloads
Article Details
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
References
Amini S., Ghobadi C., and Yamchi A. 2015. Proline accumulation and osmotic stress: an overview of P5CS gene in plants. Journal of Plant Molecular Breeding, 3(2): 44-55.
Arbona V., Manzi M., Ollas Cd. and Gómez-Cadenas A. 2013. Metabolomics as a tool to investigate abiotic stress tolerance in plants. Int J Mol Sci., 14(3): 4885-911. https://doi.org/10.3390/ijms14034885
Arriagada O., Cacciuttolo F., Cabeza R.A., Carrasco B. and Schwember A.R. 2022. A Comprehensive Review on Chickpea (Cicer arietinum L.) Breeding for Abiotic Stress Tolerance and Climate Change Resilience. International Journal of Molecular Sciences, 23(12): 6794. https://doi.org/10.3390/ijms23126794
Bates L.S., Waldren R.P.A. and Teare I.D. 1973. Rapid determination of free proline for water-stress studies. Plant and soil, 39: 205-207.
Cao X. et al. 2020. Abscisic acid mediated proline biosynthesis and antioxidant ability in roots of two different rice genotypes under hypoxic stress. BMC Plant Biol., 20: 198. https://doi.org/10.1186/s12870-020-02414-3
Chen C., Chen H., Zhang Y., Thomas H.R., Frank M.H., He Y. and Xia R. 2020. TBtools: an integrative toolkit developed for interactive analyses of big biological data. Molecular plant, 13(8): 1194-1202.
de Lima L.A.D.C., Schuster I., da Costa A.C.T. and Vendruscolo E.C. 2019. Evaluation of wheat events transformed with the p5cs gene under conditions of water stress. Revista de Ciências Agrárias, 42(2): 448-455.
Deng G., Liang J., Xu D., Long H., Pan Z. and Yu M. 2013. The relationship between proline content, the expression level of P5CS (Δ 1-pyrroline-5-carboxylate synthetase), and drought tolerance in Tibetan hulless barley (Hordeum vulgare var. nudum). Russian Journal of Plant Physiology, 60: 693-700.
El Moukhtari A., Cabassa-Hourton C., Farissi M. and Savouré A. 2020. How Does Proline Treatment Promote Salt Stress Tolerance During Crop Plant Development? Front. Plant Sci., 11:1127. https://doi.org/10.3389/fpls.2020.01127
Guan C., Cui X., Liu H-y., Li X., Li M-q. and Zhang Y-w. 2020. Proline Biosynthesis Enzyme Genes Confer Salt Tolerance to Switchgrass (Panicum virgatum L.) in Cooperation With Polyamines Metabolism. Front. Plant Sci., 11:46. https://doi.org/10.3389/fpls.2020.00046
Hayat S., Hayat Q., Alyemeni M.N., Wani A.S., Pichtel J. and Ahmad A. 2012. Role of proline under changing environments: a review. Plant signaling & behaviour, 7(11): 1456-1466.
Hosseinifard M., Stefaniak S., Ghorbani Javid M., Soltani E., Wojtyla Ł. and Garnczarska M. 2022. Contribution of Exogenous Proline to Abiotic Stresses Tolerance in Plants: A Review. International Journal of Molecular Sciences, 23(9): 5186. https://doi.org/10.3390/ijms23095186
Huang H., Ullah F., Zhou D-X., Yi M. and Zhao Y. 2019. Mechanisms of ROS Regulation of Plant Development and Stress Responses. Front. Plant Sci., 10:800. https://doi.org/10.3389/fpls.2019.00800
Hussain S., Hafeez M.B., Azam R., Mehmood K., Aziz M., Ercisli S., Javed T., Raza A., Zahra N., Hussain S. and Ren X. 2024. Deciphering the role of phytohormones and osmolytes in plant tolerance against salt stress: Implications, possible cross-talk, and prospects. Journal of Plant Growth Regulation, 43(1): 38-59.
Jain S.K., Wettberg EJv., Punia S.S., Parihar A.K., Lamichaney A., Kumar J., Gupta D.S., Ahmad S., Pant N.C., Dixit G.P., Sari H., Sari D., Ma’ruf A., Toker P. and Toker C. 2023. Genomic-Mediated Breeding Strategies for Global Warming in Chickpeas (Cicer arietinum L.). Agriculture, 13(9): 1721. https://doi.org/10.3390/agriculture13091721
Karthikeyan A., Pandian S. K. and Ramesh M. 2011. Transgenic indica rice cv. ADT 43 expressing a Δ 1-pyrroline-5-carboxylate synthetase (P5CS) gene from Vigna aconitifolia demonstrates salt tolerance. Plant Cell, Tissue and Organ Culture (PCTOC), 107: 383-395.
Kiran Kumar Ghanti S., Sujata K.G., Vijay Kumar B.M., Nataraja Karba N., Janardhan Reddy K., Srinath Rao M. and Kavi Kishor P.B. 2011. Heterologous expression of P5CS gene in chickpea enhances salt tolerance without affecting yield. Biologia Plantarum, 55: 634-640.
Koenigshofer H. and Loeppert H.G. 2019. The up-regulation of proline synthesis in the meristematic tissues of wheat seedlings upon short-term exposure to osmotic stress. Journal of plant physiology, 237: 21-29.
Liang X., Zhang L., Natarajan S.K. and Becker D.F. 2013. Proline mechanisms of stress survival. Antioxid Redox Signal., 19(9): 998-1011. https://doi.org/10.1089/ars.2012.5074.
Livak K.J. and Schmittgen T.D. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2− ΔΔCT method. Methods, 25(4): 402-408.
Maghsoudi K., Emam Y., Niazi A., Pessarakli M. and Arvin M.J. 2018. P5CS expression level and proline accumulation in the sensitive and tolerant wheat cultivars under control and drought stress conditions in the presence/absence of silicon and salicylic acid. Journal of Plant Interactions, 13(1): 461-471.
Meena M., Divyanshu K., Kumar S., Swapnil P., Zehra A., Shukla V., Yadav M. and Upadhyay R.S. 2019. Regulation of L-proline biosynthesis, signal transduction, transport, accumulation and its vital role in plants during variable environmental conditions. Heliyon, 5(12): e02952. https://doi.org/10.1016/j.heliyon.2019.e02952
Mehrotra S., Dimkpa C.O. and Goyal V. 2023. Survival mechanisms of chickpea (Cicer arietinum) under saline conditions. Plant Physiology and Biochemistry, 108168.
Panahirad S., Morshedloo M.R., Ali S., Hano C. and Kulak M., 2023. Secondary metabolites and their potential roles in plant tolerance against abiotic and biotic stress. Plant Stress, 100292.
Panigrahi S. et al. 2024. Meta QTL analysis for dissecting abiotic stress tolerance in chickpea. BMC Genomics, 25: 439. https://doi.org/10.1186/s12864-024-10336-9.
Per T.S., Khan N.A., Reddy P.S., Masood A., Hasanuzzaman M., Khan M.I.R. and Anjum N.A. 2017. Approaches in modulating proline metabolism in plants for salt and drought stress tolerance: Phytohormones, mineral nutrients and transgenics. Plant physiology and biochemistry, 115:126-140.
Rai A.N. and Penna S. 2013. Molecular evolution of plant P5CS gene involved in proline biosynthesis. Molecular Biology Reports, 40: 6429-6435.
Renzetti M., Funck D. and Trovato M. 2024. Proline and ROS: A Unified Mechanism in Plant Development and Stress Response?. Plants, 14(1), 2.
Sadiqov S.T., Akbulut M.İ.K.A.İ.L. and Ehmedov V. 2002. Role of Ca 2+ in drought stress signaling in wheat seedlings. Biochemistry (Moscow), 67: 491-497.
Sharma A., Shahzad B., Kumar V., Kohli S.K., Sidhu G.P.S., Bali A.S., Handa N., Kapoor D., Bhardwaj R. and Zheng B. 2019. Phytohormones Regulate Accumulation of Osmolytes Under Abiotic Stress. Biomolecules, 9(7): 285. https://doi.org/10.3390/biom9070285.
Surekha C.H., Kumari K.N., Aruna L.V., Suneetha G., Arundhati A. and Kavi Kishor P.B. 2014. Expression of the Vigna aconitifolia P5CSF129A gene in transgenic pigeonpea enhances proline accumulation and salt tolerance. Plant Cell, Tissue and Organ Culture (PCTOC), 116: 27-36.
Wei T.L., Wang Z.X., He Y.F., Xue S., Zhang S.Q., Pei M.S., Liu H.N., Yu Y.H. and Guo D.L. 2022. Proline synthesis and catabolism-related genes synergistically regulate proline accumulation in response to abiotic stresses in grapevines. Scientia Horticulturae, 305: 111373.
Wei T.L., Wang Z.X., He Y.F., Xue S., Zhang S.Q., Pei M.S. and Guo D.L. 2022. Proline synthesis and catabolism-related genes synergistically regulate proline accumulation in response to abiotic stresses in grapevines. Scientia Horticulturae, 305: 111373.
Yadav S, Yadava YK, Meena S, Kalwan G, Bharadwaj C, Paul V, Kansal R, Gaikwad K, and Jain P.K. 2024. Novel insights into drought-induced regulation of ribosomal genes through DNA methylation in chickpea. Int J Biol Macromol, 266(Pt2): 131380. https://doi.org/10.1016/j.ijbiomac.2024.131380
Zarattini M. and Forlani G. 2017. Toward Unveiling the Mechanisms for Transcriptional Regulation of Proline Biosynthesis in the Plant Cell Response to Biotic and Abiotic Stress Conditions. Front. Plant Sci., 8: 927. https://doi.org/10.3389/fpls.2017.00927.
Zhang C.S., Lu Q. and Verma D.P.S. 1997. Characterization of Δ1-pyrroline-5-carboxylate synthetase gene promoter in transgenic Arabidopsis thaliana subjected to water stress. Plant Science, 129(1): 81-89.
Zhang J., Wang J., Zhu C., Singh R.P. and Chen W. 2024. Chickpea: Its Origin, Distribution, Nutrition, Benefits, Breeding, and Symbiotic Relationship with Mesorhizobium Species. Plants, 13(3): 429. https://doi.org/10.3390/plants13030429.