Abstract
Key message
The content of flavonoids could increase in A. canescens under saline conditions. Overexpression of AcCHI in transgenic A. thaliana promotes flavonoid biosynthesis, thereby functioning in the tolerance of transgenic plants to salt and osmotic stress by maintaining ROS homeostasis.
Abstract
Atriplex canescens is a halophytic forage shrub with excellent adaptation to saline environment. Our previous study showed that a large number of genes related to the biosynthesis of flavonoids in A. canescens were significantly up-regulated by NaCl treatments. However, it remains unclear whether flavonoids are involved in A. canescens response to salinity. In this study, we found that the accumulation of flavonoids significantly increased in either the leaves or roots of A. canescens seedling under 100 and 300 mM NaCl treatments. Correspondingly, AcCHS, AcCHI and AcF3H, which encode three key enzymes (chalcone synthases (CHS), chalcone isomerase (CHI), and flavanone 3-hydroxylase (F3H), respectively) of flavonoids biosynthesis, were significantly induced in the roots or leaves of A. canescens by 100 or 300 mM NaCl. Then, we generated the transgenic Arabidopsis thaliana overexpressing AcCHI and found that transgenic plants accumulated more flavonoids through enhancing the pathway of flavonoids biosynthesis. Furthermore, overexpression of AcCHI conferred salt and osmotic stress tolerance in transgenic A. thaliana. Contrasted with wild-type A. thaliana, transgenic lines grew better with greater biomass, less H2O2 content as well as lower relative plasma permeability in either salt or osmotic stress conditions. In conclusion, our results indicate that flavonoids play an important role in A. canescens response to salt stress through reactive oxygen species (ROS) scavenging and the key enzyme gene AcCHI in flavonoids biosynthesis pathway of A. canescens has the potential to improve the stress tolerance of forages and crops.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data availability
All data presented in this research are available in the article and supplementary materials.
References
Agati G, Tattini M (2010) Multiple functional roles of flavonoids in photoprotection. New Phytol 186:786–793. https://doi.org/10.1111/j.1469-8137.2010.03269.x
Amorim LB, Ferreira-Neto JRC, Bezerra-Neto JP, Pandolfi V, de Araújo FT, da Silva Matos MK, Santos MG, Kido EA, Benko-Iseppon AM (2018) Cowpea and abiotic stresses: identification of reference genes for transcriptional profiling by qPCR. Plant Methods 14:88. https://doi.org/10.1186/s13007-018-0354-z
Bao AK, Wang YW, Xi JJ, Liu C, Zhang JL, Wang SM (2014) Co-expression of xerophyte Zygophyllum xanthoxylum ZxNHX and ZxVP1-1 enhances salt and drought tolerance in transgenic Lotus corniculatus L. by increasing cations accumulation. Funct Plant Biol 41:203–214. https://doi.org/10.1071/FP13106
Bhatt A, Santo A (2016) Germination and recovery of heteromorphic seeds of Atriplex canescens (Amaranthaceae) under increasing salinity. Plant Ecol 217:1069–1079. https://doi.org/10.1007/s11258-016-0633-6
Chen L, Guo H, Lin Y, Cheng H (2015) Chalcone synthase EaCHS1 from Eupatorium adenophorum functions in salt stress tolerance in tobacco. Plant Cell Rep 34:885–894. https://doi.org/10.1007/s00299-015-1751-7
Chen S, Wu F, Li Y, Qian Y, Pan X, Li F, Wang Y, Wu Z, Fu C, Lin H, Yang A (2019) NtMYB4 and NtCHS1 are critical factors in the regulation of flavonoid biosynthesis and are involved in salinity responsiveness. Front Plant Sci 10:178. https://doi.org/10.3389/fpls.2019.00178
Chmittgen T, Livak K (2008) Analyzing real-time PCR data by the comparative CT method. Nat Protoc 3:1101–1108. https://doi.org/10.1038/nprot.2008.73
Clough SJ, Bent AF (1998) Floral dip: a simplified method for Agrobacterium mediated transformation of Arabidopsis thaliana. Plant J 16:735–743. https://doi.org/10.1046/j.1365-313x.1998.00343.x
Daudi A, O’Brien JA (2012) Detection of hydrogen peroxide by DAB staining in Arabidopsis leaves. Bio Protoc 2:263–267. https://doi.org/10.21769/BIOPROTOC.263
El-Agha DE, Molle F, Rap E, Bialy ME, El-Hassan WA (2020) Drainage water salinity and quality across nested scales in the Nile Delta of Egypt. Environ Sci Pollut Res Int 27:32239–32250. https://doi.org/10.1007/s11356-019-07154-y
Feng YN, Cui JQ, Zhou T, Liu Y, Yue CP, Huang JY, Hua YP (2020) Comprehensive dissection into morpho-physiologic responses, ionomic homeostasis, and transcriptomic profiling reveals the systematic resistance of allotetraploid rapeseed to salinity. BMC Plant Biol 20:534. https://doi.org/10.1186/s12870-020-02734-4
Feng S, Wang B, Li C, Bao AK (2023) Transcriptomic analysis provides insight into the ROS scavenging system and regulatory mechanisms in Atriplex canescens response to salinity. Int J Mol Sci 24:242. https://doi.org/10.3390/ijms24010242
Flowers TJ (2004) Improving crop salt tolerance. J Exp Bot 55:307–319. https://doi.org/10.1093/jxb/erh003
Flowers TJ, Colmer TD (2015) Plant salt tolerance: adaptations in halophytes. Ann Bot 115:327–331. https://doi.org/10.1093/aob/mcu267
Flowers TJ, Rana M, Colmer TD (2015) Sodium chloride toxicity and the cellular basis of salt tolerance in halophytes. Ann Bot 3:419–431. https://doi.org/10.1093/aob/mcu217
Formentin E, Sudiro C, Ronci MB, Locato V, Barizza E, Stevanato P, Ijaz B, Zottini M, De Gara L, Lo Schiavo F (2018) H2O2 signature and innate antioxidative profile make the difference between sensitivity and tolerance to salt in rice cells. Front Plant Sci 9:1549. https://doi.org/10.3389/fpls.2018.01549
Gao JJ, Zhang Z, Peng RH, Xiong AS, Jing Xu, Zhu B, Yao QH (2011) Forced expression of Mdmyb10, a MYB transcription factor gene from apple, enhances tolerance to osmotic stress in transgenic Arabidopsis. Mol Biol Rep 38:205–211. https://doi.org/10.1007/s11033-010-0096-0
Gharibi S, Sayed Tabatabaei BE, Saeidi G, Talebi M, Matkowski A (2019) The effect of drought stress on polyphenolic compounds and expression of flavonoid biosynthesis related genes in Achillea pachycephala Rech.f. Phytochemistry 162:90–98. https://doi.org/10.1016/j.phytochem.2019.03.004
Guo H, Zhang L, Cui YN, Wang SM, Bao AK (2019) Identification of candidate genes related to salt tolerance of the secretohalophyte Atriplex canescens by transcriptomic analysis. BMC Plant Biol 19:213. https://doi.org/10.1186/s12870-019-1827-6
Guo H, Cui YN, Pan YQ, Wang SM, Bao AK (2020) Sodium chloride facilitates the secretohalophyte Atriplex canescens adaptation to drought stress. Plant Physiol Biochem 150:99–108. https://doi.org/10.1016/j.plaphy.2020.02.018
Guo H, Cui YN, Zhang L, Feng S, Ren ZJ, Wang SM, Bao AK (2022) AcHKT1; 2 is a candidate transporter mediating the influx of Na+ into the salt bladder of Atriplex canescens. Plant Soil 483:607–624. https://doi.org/10.1007/s11104-022-05769-8
Hao GY, Lucero ME, Sanderson SC, Zacharias EH, Holbrook NM (2013) Polyploidy enhances the occupation of heterogeneous environments through hydraulic related trade-offs in Atriplex canescens (Chenopodiaceae). New Phytol 197:970–978. https://doi.org/10.1111/nph.12051
Hodaei M, Rahimmalek M, Arzani A, Talebi M (2018) The effect of water stress on phytochemical accumulation, bioactive compounds and expression of key genes involved in flavonoid biosynthesis in Chrysanthemum morifolium L. Ind Crop Prod 120:295–304. https://doi.org/10.1016/j.indcrop.2018.04.073
Hossain MN, Sarker U, Raihan MS, Al-Huqail AA, Siddiqui MH, Oba S (2022) Influence of salinity stress on color parameters, leaf pigmentation, polyphenol and flavonoid contents, and antioxidant activity of Amaranthus lividus leafy vegetables. Molecules 27:1821. https://doi.org/10.3390/molecules27061821
Ijaz R, Ejaz J, Gao S, Liu T, Imtiaz M, Ye Z, Wang T (2017) Overexpression of annexin gene AnnSp2, enhances drought and salt tolerance through modulation of ABA synthesis and scavenging ROS in tomato. Sci Rep 7:12087. https://doi.org/10.1038/s41598-017-11168-2
Jayaraman K, Raman VK, Sevanthi AM, Sivakumar SR, Gayatri Viswanathan C, Trilochan M, Mandal PK (2021) Stress-inducible expression of chalcone isomerase2 gene improves accumulation of flavonoids and imparts enhanced abiotic stress tolerance to rice. Environ Exp Bot 190:104582. https://doi.org/10.1016/j.envexpbot.2021.104582
Jez JM, Bowman ME, Dixon RA, Noel JP (2000) Structure and mechanism of the evolutionarily unique plant enzyme chalcone isomerase. Nat Struct Biol 7:786–791. https://doi.org/10.1038/79025
Kim J, Lee WJ, Vu TT, Jeong CY, Hong SW, Lee H (2017) High accumulation of anthocyanins via the ectopic expression of AtDFR confers significant salt stress tolerance in Brassica napus L. Plant Cell Rep 36:1215–1224. https://doi.org/10.1007/s00299-017-2147-7
Kotula L, Garcia Caparros P, Zörb C, Colmer TD, Flowers TJ (2020) Improving crop salt tolerance using transgenic approaches: an update and physiological analysis. Plant Cell Environ 43:2932–2956. https://doi.org/10.1111/pce.13865
Li S, Sun L, Bai L, Wang W, Wang J (2017) Flavonoid is associated with salt stress tolerance in Atriplex centralasiatica seedlings. Chin J Ecol 25:1345–1350. https://doi.org/10.13930/j.cnki.cjea.170079
Li J, Hossain MS, Ma H, Yang QH, Gong XW, Yang P, Feng BL (2020a) Comparative metabolomics reveals differences in flavonoid metabolites among different coloured buckwheat flowers. J Food Compos Anal 85:103335. https://doi.org/10.1016/j.jfca.2019.103335
Li S, Chang Y, Li B, Shao SL, Zhen ZZ (2020b) Functional analysis of 4-coumarate: CoA ligase from Dryopteris fragrans in transgenic tobacco enhances lignin and flavonoids. Genet Mol Biol 43:e20180355. https://doi.org/10.1590/1678-4685-GMB-2018-0355
Liu H, Su B, Zhang H, Gong J, Zhang B, Liu Y, Du L (2019) Identification and functional analysis of a flavonol synthase gene from grape hyacinth. Molecules 24:1579. https://doi.org/10.3390/molecules24081579
Liu Y, Dai XB, Zhao LK, Huo KS, Jin PF, Zhao DL, Zhou ZL, Tang J, Xiao SZ, Cao QH (2020) RNA-seq reveals the salt tolerance of Ipomoea pes-caprae, a wild relative of sweet potato. J Plant Physiol 255:153276. https://doi.org/10.1016/j.jplph.2020.153276
Ma Q, Yue LJ, Zhang JL, Wu GQ, Bao AK, Wang SM (2012) Sodium chloride improves photosynthesis and water status in the succulent xerophyte Zygophyllum xanthoxylum. Tree Physiol 32:4–13. https://doi.org/10.1093/treephys/tpr098
Ma D, Sun D, Wang C, Li Y, Guo T (2014) Expression of flavonoid biosynthesis genes and accumulation of flavonoid in wheat leaves in response to drought stress. Plant Physiol Biochem 80:60–66. https://doi.org/10.1016/j.plaphy.2014.03.024
Ma S, Lv L, Meng C, Zhang C, Li Y (2020) Integrative analysis of the metabolome and transcriptome of Sorghum bicolor reveals dynamic changes in flavonoids accumulation under saline-alkali stress. J Agric Food Chem 68:14781–14789. https://doi.org/10.1021/acs.jafc.0c06249
Ma D, He Z, Bai X, Wang W, Zhao P, Lin P, Zhou H (2022) Atriplex canesens, a valuable plant in soil rehabilitation and forage production. A review. Sci Total Environ 804:150287. https://doi.org/10.1016/j.scitotenv.2021.150287
Mahajan M, Yadav SK (2014) Overexpression of a tea flavanone 3-hydroxylase gene confers tolerance to salt stress and Alternaria solani in transgenic tobacco. Plant Mol Biol 85:551–573. https://doi.org/10.1007/s11103-014-0203-z
Meng C, Zhang S, Deng YS, Wang GD, Kong FY (2015) Overexpression of a tomato flavanone 3-hydroxylase-like protein gene improves chilling tolerance in tobacco. Plant Physiol Biochem 96:388–400. https://doi.org/10.1016/j.plaphy.2015.08.019
Murphy A, Peer WA, Taiz L (2000) Regulation of auxin transport by aminopeptidases and endogenous flavonoids. Planta 211:315–324. https://doi.org/10.1007/s004250000300
Nabavi SM, Samec D, Tomczyk M, Milella L, Russo D, Habtemariam S, Suntar I, Rastrelli L, Daglia M, Xiao J, Giampieri F, Battino M, Sobarzo-Sanchez E, Nabavi SF, Yousefi B, Jeandet P, Xu S, Shirooie S (2020) Flavonoid biosynthetic pathways in plants: versatile targets for metabolic engineering. Biotechnol Adv 38:107316. https://doi.org/10.1016/j.biotechadv.2018.11.005
Ngaki MN, Louie GV, Philippe RN, Manning G, Pojer F, Bowman ME, Li L, Larsen E, Wurtele ES, Noel JP (2012) Evolution of the chalcone-isomerase fold from fatty-acid binding to stereospecific catalysis. Nature 485:530–533. https://doi.org/10.1038/nature11009
Ni J, Ruan R, Wang L, Jiang Z, Gu X, Chen L, Xu M (2020a) Functional and correlation analyses of dihydroflavonol-4-reductase genes indicate their roles in regulating anthocyanin changes in Ginkgo biloba. Ind Crop Prod 152:112546. https://doi.org/10.1016/j.indcrop.2020.112546
Ni R, Zhu TT, Zhang XS, Wang PY, Sun CJ, Qiao YN, Lou HX, Cheng AX (2020b) Identification and evolutionary analysis of chalcone isomerase-fold proteins in ferns. J Exp Bot 71:290–304. https://doi.org/10.1093/jxb/erz425
Pan YQ, Guo H, Wang SM, Zhao B, Zhang JL, Ma Q, Yin HJ, Bao AK (2016) The photosynthesis, Na+/K+ homeostasis and osmotic adjustment of Atriplex canescens in response to salinity. Front Plant Sci 7:848. https://doi.org/10.3389/fpls.2016.00848
Sarker U, Ercisli S (2022) Salt eustress induction in red Amaranth (Amaranthus gangeticus) augments nutritional, phenolic acids and antiradical potential of leaves. Antioxidants (Basel) 11:2434. https://doi.org/10.3390/antiox11122434
Sarker U, Oba S (2018) Augmentation of leaf color parameters, pigments, vitamins, phenolic acids, flavonoids and antioxidant activity in selected Amaranthus tricolor under salinity stress. Sci Rep 8:12349. https://doi.org/10.1038/s41598-018-30897-6
Sarker U, Oba S (2019) Salinity stress enhances color parameters, bioactive leaf pigments, vitamins, polyphenols, flavonoids and antioxidant activity in selected Amaranthus leafy vegetables. J Sci Food Agric 99:2275–2284. https://doi.org/10.1002/jsfa.9423
Sarker U, Oba S (2020) The response of salinity stress-induced A. tricolor to growth, anatomy, physiology, non-enzymatic and enzymatic antioxidants. Front Plant Sci 11:559876. https://doi.org/10.3389/fpls.2020.559876
Sarker U, Islam MT, Oba S (2018) Salinity stress accelerates nutrients, dietary fiber, minerals, phytochemicals and antioxidant activity in Amaranthus tricolor leaves. PLoS ONE 13:e0206388
Sarker U, Hossain MN, Oba S, Ercisli S, Marc RA, Golokhvast KS (2023) Salinity stress ameliorates pigments, minerals, polyphenolic profiles, and antiradical capacity in Lalshak. Antioxidants (Basel) 12:173. https://doi.org/10.3390/antiox12010173
Sewelam N, Kazan K, Schenk PM (2016) Global plant stress signaling: reactive oxygen species at the cross-road. Front Plant Sci 7:187. https://doi.org/10.3389/fpls.2016.00187
Shabala S (2013) Learning from halophytes: physiological basis and strategies to improve abiotic stress tolerance in crops. Ann Bot 7:1209–1221. https://doi.org/10.1093/aob/mct205
Shimada N, Aoki T, Sato S, Nakamura Y, Tabata S, Ayabe S (2003) A cluster of genes encodes the two types of chalcone isomerase involved in the biosynthesis of general flavonoids and legume-specific 5-deoxy(iso)flavonoids in Lotus japonicus. Plant Physiol 131:941–951. https://doi.org/10.1104/pp.004820
Singh A (2018) Alternative management options for irrigation-induced salinization and waterlogging under different climatic conditions. Ecol Indic 90:184–192. https://doi.org/10.1016/j.ecolind.2018.03.014
Sun W, Shen H, Xu H, Tang X, Tang M, Ju Z, Yi Y (2019) Chalcone isomerase a key enzyme for anthocyanin biosynthesis in Ophiorrhiza japonica. Front Plant Sci 10:865. https://doi.org/10.3389/fpls.2019.00865
Wang H, Hu T, Huang J, Lu X, Huang B, Zheng Y (2013) The expression of Millettia pinnata chalcone isomerase in Saccharomyces cerevisiae salt-sensitive mutants enhances salt-tolerance. Int J Mol Sci 14:8775–8786. https://doi.org/10.3390/ijms14058775
Wang F, Ren G, Li F, Qi S, Xu Y, Wang B, Yang Y, Ye Y, Zhou Q, Chen X (2018) A chalcone synthase gene AeCHS from Abelmoschus esculentus regulates flavonoid accumulation and abiotic stress tolerance in transgenic Arabidopsis. Acta Physiol Plant 40:97. https://doi.org/10.1007/s11738-018-2680-1
Wang H, Liu S, Wang T, Liu H, Xu X, Chen K, Zhang P (2020a) The moss flavone synthase I positively regulates the tolerance of plants to drought stress and UV-B radiation. Plant Sci 298:110591. https://doi.org/10.1016/j.plantsci.2020.110591
Wang N, Liu W, Yu L, Guo Z, Chen Z, Jiang S, Xu H, Fang H, Wang Y, Zhang Z, Chen X (2020b) HEAT SHOCK FACTOR A8a modulates flavonoid synthesis and drought tolerance. Plant Physiol 184:1273–1290. https://doi.org/10.1104/pp.20.01106
Wang M, Ren T, Huang R, Li Y, Zhang C, Xu Z (2021) Overexpression of an Apocynum venetum flavonols synthetase gene confers salinity stress tolerance to transgenic tobacco plants. Plant Physiol Biochem 162:667–676. https://doi.org/10.1016/j.plaphy.2021.03.034
Xu N, Liu S, Lu Z, Pang S, Wang L, Wang L, Li W (2020) Gene expression profiles and flavonoid accumulation during salt stress in Ginkgo biloba seedlings. Plants 9:1162. https://doi.org/10.3390/plants9091162
Yang Y, Guo Y (2018) Unraveling salt stress signaling in plants. J Integr Plant Biol 60:796–804. https://doi.org/10.1111/jipb.12689
Yang L, Zhang JC, Qu JT, He G, Yu HQ, Li WC, Fu FL (2019) Expression response of chalcone synthase gene to inducing conditions and its effect on flavonoids accumulation in two medicinal species of Anoectochilus. Sci Rep 9:20171. https://doi.org/10.1038/s41598-019-56821-0
Zhang JL, Shi H (2013) Physiological and molecular mechanisms of plant salt tolerance. Photosynth Res 115:1–22. https://doi.org/10.1007/s11120-013-9813-6
Zhang H, Han B, Wang T, Chen S, Li H, Zhang Y, Dai S (2012) Mechanisms of plant salt response: insights from proteomics. J Proteome Res 11:49–67. https://doi.org/10.1021/pr200861w
Zhang K, Sun Y, Li M, Long R (2021) CrUGT87A1, a UDP-sugar glycosyltransferases (UGTs) gene from Carex rigescens, increases salt tolerance by accumulating flavonoids for antioxidation in Arabidopsis thaliana. Plant Physiol Biochem 159:28–36. https://doi.org/10.1016/j.plaphy.2020.12.006
Zhang H, Zhu J, Gong Z, Zhu JK (2022) Abiotic stress responses in plants. Nat Rev Genet 23:104–119. https://doi.org/10.1038/s41576-021-00413-0
Zhao C, Liu X, Gong Q, Cao J, Shen W, Yin X, Grierson D, Zhang B, Xu C, Li X, Chen K, Sun C (2021) Three AP2/ERF family members modulate flavonoid synthesis by regulating type IV chalcone isomerase in citrus. Plant Biotechnol J 19:671–688. https://doi.org/10.1111/pbi.13494
Zheng XT, Chen YL, Zhang XH, Cai ML, Yu ZC, Peng CL (2019) ANS-deficient Arabidopsis is sensitive to high light due to impaired anthocyanin photoprotection. Funct Plant Biol 46:756–765. https://doi.org/10.1071/fp19042
Zhou L, Wang Y, Ren L, Shi QQ, Zheng BQ, Miao K, Guo X (2014) Overexpression of Ps-CHI1, a homologue of the chalcone isomerase gene from tree peony (Paeonia suffruticosa), reduces the intensity of flower pigmentation in transgenic tobacco. Plant Cell Tissue Organ Cult 116:285–295. https://doi.org/10.1007/s11240-013-0403-2
Zhu J, Zhao W, Li R, Guo D, Li H, Wang Y, Mei W, Peng S (2021) Identification and characterization of chalcone isomerase genes involved in flavonoid production in Dracaena cambodiana. Front Plant Sci 12:616396. https://doi.org/10.3389/fpls.2021.616396
Acknowledgements
We thank for the editors and anonymous reviewers for their constructive suggestions.
Funding
This work was supported by the National Natural Science Foundation of China (31971761, 32371756), the Key Science & Technology Project of Gansu Province (22ZD6NA007), the Science & Technology Project from China Huaneng Group Co. LTD. (HNKJ21-H76), and the Fundamental Research Funds for the Central Universities (lzujbky-2022-ct03).
Author information
Authors and Affiliations
Contributions
A-KB conceived and designed the study. SF, Y-TY, and B-BW performed the experiments. SF wrote the manuscript. SF, Y-ML, LL, and A-KB revised the manuscript. All author read and approved the final manuscript.
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
Additional information
Communicated by Li Tian.
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
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
Feng, S., Yao, YT., Wang, BB. et al. Flavonoids are involved in salt tolerance through ROS scavenging in the halophyte Atriplex canescens. Plant Cell Rep 43, 5 (2024). https://doi.org/10.1007/s00299-023-03087-6
Received:
Accepted:
Published:
DOI: https://doi.org/10.1007/s00299-023-03087-6