Abstract
Rising global temperatures are expected to worsen the impact of drought, which directly impacts the worldwide production of chickpea. Glutaredoxins (Grxs) are ubiquitous, heat-stable, cysteine-rich proteins and glutathione-dependent thiol-disulfide oxidoreductases. The overexpression of the CC-type Grxs gene in plants improves drought and salinity stress tolerance. The present study reports the potential role of chickpea glutaredoxin (CaGrx) gene in response to drought stress. The CaGrx gene from chickpea was overexpressed in Kabuli-type chickpea by the Agrobacterium-mediated gene transformation method. Integration of the CaGrx gene in the Kabuli chickpea genome was confirmed by PCR using gene-specific primers. The qRT-PCR showed an 11- to 14-fold change in the expression of the CaGrx transcript in the T1 generation of transgenic chickpea. The T1 generation of the transgenic chickpea exhibit improved tolerance to drought stress due to the enhanced activities of different antioxidant enzymes such as glutaredoxin (GRX), glutathione reductase (GR), glutathione peroxidase (GPX), glutathione-S-transferase (GST), superoxide dismutase (SOD), and catalase (CAT), and reduced levels of stress markers H2O2 and thiobarbituric acid reactive substances (TBARS). Also, the CaGrx overexpression lines of T1 generation showed improved physiological performance, including net photosynthesis (PN), transpiration (E), water use efficiency (WUE), stomatal conductance (gs), PSII (Fv/Fm), and non-photochemical quenching (NPQ) during drought, which help to maintain the photosynthetic apparatus and protect the plants from oxidative damage. Overall, the study showed that the overexpression of CaGrx improves the ability of chickpea to withstand drought, and the CaGrx gene could be used to develop drought-tolerant crop plants.
Key message
Overexpression of the chickpea glutaredoxin (CaGrx) gene in chickpea enhances drought tolerance by enhancing antioxidant enzymes defense, reducing oxidative damage, and boosting plant growth and productivity under drought stress conditions.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data availability
The data generated and/or analyzed during the current study are available from the corresponding author upon reasonable request.
References
Abid M, Ali S, Qi LK (2018) Physiological and biochemical changes during drought and recovery periods at tillering and jointing stages in wheat (Triticum aestivum L). Sci Rep 8:4615. https://doi.org/10.1038/s41598-018-21441
Baisak R, Rana D, Acharya PB, Kar M (1994) Alterations in the activities of active oxygen scavenging enzymes of wheat leaves subjected to water stress. Plant Cell Physiol 35:489–495. https://doi.org/10.1590/S1677-04202007000100006
Beauchamp C, Fridovich I (1971) Superoxide dismutase: improved assays and an assay applicable to acrylamide gels. Anal Biochem 44:276–287. https://doi.org/10.1016/0003-2697(71)90370
Berwal M, Ram C (2018) Superoxide dismutase: a stable biochemical marker for abiotic stress tolerance in higher plants. Abiotic and Biotic Stress in Plants 1–10. https://doi.org/10.5772/intechopen.82079
Blum A (2009) Effective use of water (EUW) and not water-use efficiency (WUE) is the target of crop yield improvement under drought stress. Field Crops Research 112:119–123. https://doi.org/10.1016/j.fcr.2009.03.009
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254. https://doi.org/10.1016/0003-2697(76)90527
Chandlee JM, Scandalios JG (1984) Analysis of variants affecting the catalase developmental program in maize scutellum. Theor Appl Genet 69:71–77. https://doi.org/10.1007/BF00262543
de la Fuente Canto C, Grondin A, Sine B (2023) Glutaredoxin regulation of primary root growth confers early drought stress tolerance in pearl millet. https://doi.org/10.1101/2023.02.02.526762. BioRxiv 2023-02
Dubey AK, Kumar N, Sahu N et al (2016) Response of two rice cultivars differing in their sensitivity towards arsenic, differs in their expression of glutaredoxin and glutathione S transferase genes and antioxidant usage. Ecotoxicol Environ Saf 124:393–405. https://doi.org/10.1016/j.ecoenv.2015.10.017
Dubey R, Dudhagi SS, Narayan S et al (2022) Stomatal behaviour and endogenous phytohormones promotes intrinsic water use efficiency differently in cotton under drought. Indian J Exp Biol 60:112–120. https://doi.org/10.56042/ijeb.v60i02.42969
El-Kereamy A, Bi YM, Mahmood K et al (2015) Overexpression of the CC-type glutaredoxin, OsGRX6 affects hormone and nitrogen status in rice plants. Front Plant Sci 6:934. https://doi.org/10.3389/fpls.2015.00934
Fang X, Turner NC, Yan G, Li F, Siddique KH (2010) Flower numbers, pod production, pollen viability, and pistil function are reduced and flower and pod abortion increased in chickpea (Cicer arietinum L.) under terminal drought. J Exp Bot 61:335–345. https://doi.org/10.1093/jxb/erp307
FAO (2021) FAOSTAT: Food and Agriculture data: www.faostat.org
Gallogly MM, Mieyal JJ (2007) Mechanisms of reversible protein glutathionylation in redox signaling and oxidative stress. Curr Opin Pharmacol 7:381–391. https://doi.org/10.1016/j.coph.2007.06.003
Gaur PM, Jukanti AK, Samineni S et al (2013) Climate change and heat stress tolerance in chickpea. Clim Change Plant Abiotic Stress Tolerance 837–856. https://doi.org/10.1002/9783527675265
Gill SS, Tuteja N (2010) Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants. Plant Physiol Biochem 48:909–930. https://doi.org/10.1016/j.plaphy.2010.08.016
Habig WH, Pabst MJ, Jakoby WB (1974) Glutathione S-transferases: the first enzymatic step in mercapturic acid formation. J Biol Chem 249:7130–7139. https://doi.org/10.1016/s0021-9258(19)42083
Henckel PA (1964) Physiology of plants under drought. Annu Rev Plant Physiol 15:363–386. https://doi.org/10.1146/annurev.pp.15.060164.002051
Hodges DM, DeLong JM, Forney CF, Prange RK (1999) Improving the thiobarbituric acid-reactive-substances assay for estimating lipid peroxidation in plant tissues containing anthocyanin and other interfering compounds. Planta 207:604–611. https://doi.org/10.1007/s004250050524
Holmgren A, Aslund F (1995) Glutaredoxin. Methods Enzymol 252:283–292. https://doi.org/10.1016/0076-6879(95)52031
Jha UC, Chaturvedi SK, Bohra A, Basu PS, Khan MS, Barh D (2014) Abiotic stresses, constraints and improvement strategies in chickpea. Plant Breeding 133:163–178. https://doi.org/10.1111/pbr.12150
Kadiyala MDM, Charyulu DK, Nedumaran S et al (2016) Agronomic management options for sustaining chickpea yield under climate change scenario. J Agrometeorology 18:41–47. https://doi.org/10.54386/jam.v18i1.897
Kato Y, Hirotsu S, Nemoto K, Yamagishi J (2008) Identification of QTLs controlling rice drought tolerance at seedling stage in hydroponic culture. Euphytica 160:423–430. https://doi.org/10.1007/s10681-007-9605
Kumar A, Dubey AK, Kumar V et al (2020) Overexpression of rice glutaredoxin genes LOC_Os02g40500 and LOC_Os01g27140 regulate plant responses to drought stress. Ecotoxicology and Environmental Safety 200: 110721. https://doi.org/10.1016/j.ecoenv.2020.110721
Kumar A, Kumar V, Dubey AK et al (2021) Chickpea glutaredoxin (CaGrx) gene mitigates drought and salinity stress by modulating the physiological performance and antioxidant defense mechanisms. Physiol Mol Biology Plants 27:923–944. https://doi.org/10.1007/s12298-021-00999
Leport L, Turner NC, Davies SL, Siddique KHM (2006) Variation in pod production and abortion among chickpea cultivars under terminal drought. Eur J Agron 24:236–246. https://doi.org/10.1016/j.eja.2005.08.005
Li K, Yang F, Zhang G, Song S, Li Y, Ren D, Miao Y, Song CP (2017) AIK1, a mitogen-activated protein kinase, modulates abscisic acid responses through the MKK5-MPK6 kinase cascade. Plant Physiol 173:1391–1408. https://doi.org/10.1104/pp.16.01386
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2 – ∆∆CT method. Methods 25:402–408. https://doi.org/10.1006/meth.2001.1262
Mahmood A, Kanwal H, Kausar A et al (2019) Seed priming with zinc modulate growth, pigments and yield of chickpea (Cicer arietinum L.) under water deficit conditions. Appl Ecol Environ Res 17:147–160. https://doi.org/10.15666/aeer/1701-147160
Martin-StPaul N, Delzon S, Cochard H (2017) Plant resistance to drought depends on timely stomatal closure. Ecol Lett 20:1437–1447. https://doi.org/10.1111/ele.12851
Meenakshi KA, Kumar V et al (2022) CAMTA transcription factor enhances salinity and drought tolerance in chickpea (Cicer arietinum L). Plant Cell Tissue and Organ Culture (PCTOC) 148:319–330. https://doi.org/10.1007/s11240-021-02191-3
Mishra MK, Chaturvedi P, Singh R (2013) Overexpression of WsSGTL1 gene of Withania somnifera enhances salt tolerance, heat tolerance and cold acclimation ability in transgenic Arabidopsis plants. PLoS ONE 8:63064. https://doi.org/10.1371/journal.pone.0063064
Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with Tobacco tissue cultures. Physiol Plant 15:473–497. https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
Ozyigit II, Filiz E, Vatansever R (2016) Identification and comparative analysis of H2O2-scavenging enzymes (ascorbate peroxidase and glutathione peroxidase) in selected plants employing bioinformatics approaches. Front Plant Sci 7:301. https://doi.org/10.3389/fpls.2016.00301
Panwar V, Sharma A, Sen S (2021) Economic consequences of slow-and fast-onset Natural Disasters: empirical evidences from India. Economic Eff Nat Disasters 601–623. https://doi.org/10.1016/B978-0-12-817465-4.00035
Posgai AL, Wasserfall CH, Kwon KC (2017) Plant-based vaccines for oral delivery of type 1 diabetes-related autoantigens: evaluating oral tolerance mechanisms and Disease prevention in NOD mice. Sci Rep 7:42372. https://doi.org/10.1038/srep42372
Rouhier N, Couturier J, Jacquot JP (2006) Genome-wide analysis of plant glutaredoxin systems. J Exp Bot 57:1685–1696. https://doi.org/10.1093/jxb/erl001
Rouhier N, Lemaire SD, Jacquot JP (2008) The role of glutathione in photosynthetic organisms: emerging functions for glutaredoxins and glutathionylation. Annu Rev Plant Biol 59:143–166. https://doi.org/10.1146/annurev.arplant.59.032607.092811
Salvi P, Kamble NU, Majee M (2020) Ectopic over-expression of ABA-responsive Chickpea galactinol synthase (CaGolS) gene results in improved tolerance to dehydration stress by modulating ROS scavenging. Environ Exp Bot 171:103957. https://doi.org/10.1016/j.envexpbot.2019.103957
Sanyal I, Singh AK, Kaushik M (2005) Agrobacterium-mediated transformation of chickpea (Cicer arietinum L.) with Bacillus thuringiensis cry1Ac gene for resistance against pod borer insect Helicoverpa armigera. Plant Sci 168:1135–1146. https://doi.org/10.1016/j.plantsci.2004.12.015
Sharma I, Ahmad P (2014) Catalase: a versatile antioxidant in plants. Oxidative Damage to Plants 131–148. https://doi.org/10.1016/B978-0-12-799963-0.00004
Sharma R, Priya P, Jain M (2013) Modified expression of an auxin-responsive rice CC-type glutaredoxin gene affects multiple abiotic stress responses. Planta 238:871–884. https://doi.org/10.1007/s00425-013-1940
Singh R, Yadav R, Amla DV (2016) Characterization and functional validation of two scaffold attachment regions (SARs) from Cicer arietinum (L). Plant Cell Tissue and Organ Culture (PCTOC) 125:135–148. https://doi.org/10.1007/s11240-015-0935-8
Slatyer RO, Shmueli E (1967) Measurements of internal water status and transpiration. Irrig Agricultural Lands 11:337–353. https://doi.org/10.2134/agronmonogr11
Smith IK, Vierheller TL, Thorne CA (1989) Properties and functions of glutathione reductase in plants. Physiol Plant 77:449–456. https://doi.org/10.1111/j.1399-3054.1989.tb05666
Sofi SA, Singh J, Muzaffar, Majid D, Dar BN (2020) Physicochemical characteristics of protein isolates from native and germinated chickpea cultivars and their noodle quality. Int J Gastronomy Food Sci 22:100258. https://doi.org/10.1016/j.ijgfs.2020.100258
Soni SK, Mishra MK, Mishra M (2022) Papaya leaf curl virus (PaLCuV) Infection on papaya (Carica papaya L.) plants alters anatomical and physiological properties and reduces bioactive components. Plants 11:579. https://doi.org/10.3390/plants11050579
Turner NC (1981) Techniques and experimental approaches for the measurement of plant water status. Plant Soil 58:339–366. https://doi.org/10.1007/BF02180062
Varshney RK, Thudi M, Nayak SN et al (2014) Genetic dissection of drought tolerance in chickpea (Cicer arietinum L). Theor Appl Genet 127:445–462. https://doi.org/10.1007/s00122-013-2230-6
Velikova V, Yordanov I, Edreva AJPS (2000) Oxidative stress and some antioxidant systems in acid rain-treated bean plants: protective role of exogenous polyamines. Plant Sci 151:59–66. https://doi.org/10.1016/S0168-9452(99)00197
Wonisch W, Schaur RJ (2001) Chemistry of glutathione. Significance of glutathione to plant adaptation to the environment 13–26. https://doi.org/10.1007/0-306-47644
Young JL, Dean DA (2015) Electroporation-mediated gene delivery. Adv Genet 89:49–88. https://doi.org/10.1016/bs.adgen.2014.10.003
Zander M, Chen S, Imkampe J, Thurow C, Gatz C (2012) Repression of the Arabidopsis thaliana jasmonic acid/ethylene-induced defense pathway by TGA-interacting glutaredoxins depends on their C-terminal ALWL motif. Mol Plant 5:831–840. https://doi.org/10.1093/mp/ssr113
Zoghi Z, Hosseini SM, Kouchaksaraei MT et al (2019) The effect of biochar amendment on the growth, morphology and physiology of Quercus castaneifolia seedlings under water-deficit stress. Eur J for Res 138:967–979. https://doi.org/10.1007/s10342-019-01217
Acknowledgements
The authors are grateful to the Director, CSIR-National Botanical Research Institute, Lucknow, for infrastructural support to carry out the research. VK are also thankful to the University Grants Commission (UGC), New Delhi, for providing research fellowships. CSIR-NBRI Manuscript No. CSIR-NBRI_MS/2023/10/11.
Funding
This study was funded by the Council of Scientific and Industrial Research, India (MLP 0026).
Author information
Authors and Affiliations
Contributions
VK: Conceptualization, Methodology, Formal analysis, Investigation, Writing – original draft, Validation, AK: Formal analysis, Methodology, M: Formal analysis, Investigation, UG: Investigation, SN: Investigation, PAS: Formal analysis, IS: Formulated the original research plans, supervised the research and finalized the manuscript.
Corresponding author
Ethics declarations
Ethical approval
The article contains no studies with human participants or animals.
Conflict of interest
The authors declare that they have no known competing academic and financial interests.
Additional information
Communicated by Christell van der Vyver.
Publisher’s Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Electronic Supplementary Material
11240_2023_2651_MOESM3_ESM.png
Supplementary Figure 1: Hardening and acclimatization of CaGrx overexpression and control chickpea shoots grafted on seedling stocks for the development of transgenic and control plants (a) Control chickpea plant, and (b-c) Transgenic plants. The inset shows the region of the graft union.
11240_2023_2651_MOESM4_ESM.png
Supplementary Figure 2: The relative expression of chickpea CaGrx gene. WT represents treated wild-type plants C represents wild-type control plants. The standard error was estimated from the mean of three replicates. The estimated data were applied to a one-way ANOVA through DMRT (Duncan’s Multiple Range Test). The letters above the bars determine significant variation among the treatments at different water levels at P <0.05.
11240_2023_2651_MOESM5_ESM.png
Supplementary Figure 3: Enhanced relative water content (RWC). 'C' represents wild-type Control, ‘TC’ represents Transgenic Control, and ‘WT’ represents treated wild-type plants. The control (C) and transgenic control (TC) plants were taken under well-watered, controlled conditions. The standard error was estimated from the mean of three replicates. The estimated data were applied to a one-way ANOVA through DMRT (Duncan's Multiple Range Test). The letters above the bars determine significant variation among the treatments at different water levels at P <0.05.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Kumar, V., Kumar, A., Meenakshi et al. Overexpressing the glutaredoxin (CaGrx) gene enhances the antioxidant defences and improves drought tolerance in chickpea (Cicer arietinum L.). Plant Cell Tiss Organ Cult 156, 34 (2024). https://doi.org/10.1007/s11240-023-02651-y
Received:
Accepted:
Published:
DOI: https://doi.org/10.1007/s11240-023-02651-y