Improvement of leaf K⁺ retention is a shared mechanism behind CeO₂ and Mn₃O₄ nanoparticles improved rapeseed salt tolerance

Abstract

Salinity is a global issue limiting efficient agricultural production. Nanobiotechnology has been emerged as an effective approach to improve plant salt tolerance. However, little known is about the shared mechanisms between different nanomaterials-enabled plant salt tolerance. In this study, we found that both PNC [polyacrylic acid coated nanoceria (CeO₂ nanoparticles)] and PMO (polyacrylic acid coated Mn₃O₄ nanoparticles) nanozymes improved rapeseed salt tolerance. PNC and PMO treated rapeseed plants showed significantly fresh weight, dry weight, higher chlorophyll content, Fv/Fm, and carbon assimilation rate than control plants under salt stress. Results from confocal imaging with reactive oxygen species (ROS) fluorescent dye and histochemical staining experiments showed that the ROS over-accumulation level in PNC and PMO treated rapeseed was significantly lower than control plants under salt stress. Confocal imaging results with K⁺ fluorescent dye showed that significantly higher cytosolic and vacuolar K⁺ signals were observed in PNC and PMO treated rapeseed than control plants under salt stress. This is further confirmed by leaf K⁺ content data. Furthermore, we found that PNC and PMO treated rapeseed showed significantly lower cytosolic Na⁺ signals than control plants under salt stress. While, compared with significantly higher vacuolar Na⁺ signals in PNC treated plants, PMO treated rapeseed showed significantly lower vacuolar Na⁺ signals than control plants under salt stress. These results are further supported by qPCR results of genes of Na⁺ and K⁺ transport. Overall, our results suggest that besides maintaining ROS homeostasis, improvement of leaf K⁺ retention could be a shared mechanism in nano-improved plant salt tolerance.

Keywords

Nanomaterials / TEM imaging / Gene expression / Reactive oxygen species / K⁺/Na⁺ ratio

References

[1] AnJ, HuPG, LiFJ, WuHH, ShenY, WhiteJC, TianXL, LiZH, GiraldoJP. Emerging investigator series: molecular mechanisms of plant salinity stress tolerance improvement by seed priming with cerium oxide nanoparticles. Environ Sci Nano, 2020, 7(8):2214-2228

[2] Ankit G, Beenu T, Kumar Sihag M, Vikas K, Vivek S, Suman S (2021). Rapeseed/Canola (Brassica napus) seed. In: Tanwar B, Goyal A (eds) Oilseeds: health attributes and food applications. Springer, Singapore, pp 47–71. https://doi.org/10.1007/978-981-15-4194-0_2

[3] AnzanoA, BonanomiG, MazzoleniS, LanzottiV. Plant metabolomics in biotic and abiotic stress: a critical overview. Phytochem Rev, 2022, 21(2):503-524

[4] BarassiCA, AyraultG, CreusCM, SueldoRJ, SobreroMT. Seed inoculation with Azospirillum mitigates NaCl effects on lettuce. Sci Hortic, 2006, 109(1):8-14

[5] BoseJ, Rodrigo-MorenoA, ShabalaS. ROS homeostasis in halophytes in the context of salinity stress tolerance. J Exp Bot, 2014, 65(5):1241-1257

[6] CardenDE, WalkerDJ, FlowersTJ, MillerAJ. Single-cell measurements of the contributions of cytosolic Na⁺ and K⁺ to salt tolerance. Plant Physiol, 2003, 131(2):676-683

[7] ChenZ, NewmanI, ZhouM, MendhamN, ZhangG, ShabalaS. Screening plants for salt tolerance by measuring K⁺ flux: a case study for barley. Plant Cell Environ, 2005, 28(10):1230-1246

[8] ChenLL, PengYQ, ZhuL, HuangY, BieZL, WuHH. CeO₂ nanoparticles improved cucumber salt tolerance is associated with its induced early stimulation on antioxidant system. Chemosphere, 2022, 299: 134474

[9] CuinTA, BettsSA, ChalmandrierR, ShabalaS. A root's ability to retain K⁺ correlates with salt tolerance in wheat. J Exp Bot, 2008, 59(10):2697-2706

[10] DemidchikV. Mechanisms and physiological roles of K⁺ efflux from root cells. J Plant Physiol, 2014, 171(9):696-707

[11] Deng X, Chen Y, Yang Y, Lu L, Yuan X, Zeng H, Zeng Q (2020) Cadmium accumulation in rice (Oryza sativa L.) alleviated by basal alkaline fertilizers followed by topdressing of manganese fertilizer. Environ Pollut 262:114289. https://doi.org/10.1016/j.envpol.2020.114289

[12] GaoZ, ZhangJ, ZhangJ, ZhangW, ZhengL, BorjiginT, WangY. Nitric oxide alleviates salt-induced stress damage by regulating the ascorbate-glutathione cycle and Na⁺/K⁺ homeostasis in Nitraria tangutorum Bobr. Plant Physiol Biochem, 2022, 173: 46-58

[13] Gohari G, Zareei E, Rostami H, Panahirad S, Kulak M, Farhadi H, Amini M, Martinez-Ballesta MDC, Fotopoulos V (2021) Protective effects of cerium oxide nanoparticles in grapevine (Vitis vinifera L.) cv. Flame Seedless under salt stress conditions. Ecotoxicol Environ Saf 220:112402. https://doi.org/10.1016/j.ecoenv.2021.112402

[14] Imadi SR, Shah SW, Kazi AG, Azooz MM, Ahmad P (2016) Phytoremediation of saline soils for sustainable agricultural productivity. In: Ahmad P (ed) Plant metal interaction. Elsevier, Holland, pp 455–468. https://doi.org/10.1016/B978-0-12-803158-2.00018-7

[15] KaramiA, SepehriA. Beneficial role of MWCNTs and SNP on growth, physiological and photosynthesis performance of barley under NaCl stress. J Soil Sci Plant Nut, 2018, 18(3):752-771

[16] KhanI, RazaMA, AwanSA, ShahGA, RizwanM, AliB, TariqR, HassanMJ, AlyemeniMN, BresticM, ZhangX, AliS, HuangL. Amelioration of salt induced toxicity in pearl millet by seed priming with silver nanoparticles (AgNPs): the oxidative damage, antioxidant enzymes and ions uptake are major determinants of salt tolerant capacity. Plant Physiol Biochem, 2020, 156: 221-232

[17] Khan MN, Li YH, Fu CC, Hu J, Chen LL, Yan JS, Khan Z, Wu HH, Li ZH (2022) CeO₂ nanoparticles seed priming increases salicylic acid level and ROS scavenging ability to improve rapeseed salt tolerance. Global Chall n/a(n/a):2200025. https://doi.org/10.1002/gch2. 202200025

[18] KhanMN, LiYH, KhanZ, ChenLL, LiuJH, HuJ, WuHH, LiZH. Nanoceria seed priming enhanced salt tolerance in rapeseed through modulating ROS homeostasis and alpha-amylase activities. J Nanobiotechnology, 2021, 19(1):276

[19] KumarD, YusufMA, SinghP, SardarM, SarinNB. Modulation of antioxidant machinery in alpha-tocopherol-enriched transgenic Brassica juncea plants tolerant to abiotic stress conditions. Protoplasma, 2013, 250(5):1079-1089

[20] LiYH, LiuJH, FuCC, KhanMN, HuJ, ZhaoFM, WuHH, LiZH. CeO₂ nanoparticles modulate Cu-Zn superoxide dismutase and lipoxygenase-IV isozyme activities to alleviate membrane oxidative damage to improve rapeseed salt tolerance. Environ Sci Nano, 2022, 9(3):1116-1132

[21] LiZQ, ZhuL, ZhaoFM, LiJQ, ZhangX, KongXJ, WuHH, ZhangZY. Plant salinity stress response and nano-enabled plant salt tolerance. Front Plant Sci, 2022, 13: 843994

[22] LiuJ, ShabalaS, ShabalaL, ZhouM, MeinkeH, VenkataramanG, ChenZ, ZengF, ZhaoQ. Tissue-specific regulation of Na⁺ and K⁺ transporters explains genotypic differences in salinity stress tolerance in rice. Front Plant Sci, 2019, 10: 1361

[23] LiuJH, LiGJ, ChenLL, GuJJ, WuHH, LiZH. Cerium oxide nanoparticles improve cotton salt tolerance by enabling better ability to maintain cytosolic K⁺/Na⁺ ratio. J Nanobiotechnology, 2021, 19(1):153

[24] LiuY, CaoX, YueL, WangC, TaoM, WangZ, XingB. Foliar-applied cerium oxide nanomaterials improve maize yield under salinity stress: reactive oxygen species homeostasis and rhizobacteria regulation. Environ Pollut, 2022, 299: 118900

[25] LivakKJ, SchmittgenTD. Analysis of relative gene expression data using real-time quantitative PCR and the 2^−ΔΔCT method. Methods, 2001, 25(4):402-408

[26] LuL, HuangM, HuangYX, CorviniPFX, JiR, ZhaoLJ. Mn₃O₄ nanozymes boost endogenous antioxidant metabolites in cucumber (Cucumis sativus) plant and enhance resistance to salinity stress. Environ Sci Nano, 2020, 7(6):1692-1703

[27] MejicanosG, SanjayanN, KimIH, NyachotiCM. Recent advances in canola meal utilization in swine nutrition. J Anim Sci Technol, 2016, 58: 7

[28] MittlerR. ROS are good. Trends Plant Sci, 2017, 22(1):11-19

[29] Mohammadi MHZ, Panahirad S, Navai A, Bahrami MK, Kulak M, Gohari G (2021) Cerium oxide nanoparticles (CeO₂-NPs) improve growth parameters and antioxidant defense system in Moldavian Balm (Dracocephalum moldavica L.) under salinity stress. Plant Stress https://doi.org/10.1016/j.stress.2021.100006

[30] MunnsR, TesterM. Mechanisms of salinity tolerance. Annu Rev Plant Bio, 2008, 59(5):651-681

[31] NewkirkGM, WuH, SantanaI, GiraldoJP. Catalytic scavenging of plant reactive oxygen species in vivo by anionic cerium oxide nanoparticles. J vis Exp, 2018, 26(138):58373

[32] OliveiraHC, GomesBC, PelegrinoMT, SeabraAB. Nitric oxide-releasing chitosan nanoparticles alleviate the effects of salt stress in maize plants. Nitric Oxide, 2016, 61: 10-19

[33] Pace R, Benincasa P, Ghanem ME, Quinet M, Lutts S (2012) Germination of untreated and primed seeds in rapeseed (Brassica napus L.) under salinity and low matric potential. Exp Agric 48(2):238–251. https://doi.org/10.1017/S0014479711001189

[34] RashedMH, HoqueTS, JahangirMMR, HashemMA. Manganese as a micronutrient in agriculture: crop requirement and management. J Environ Sci & Natural Resources, 2019, 12(1&2):225-241

[35] RossiL, ZhangW, LombardiniL, MaXM. The impact of cerium oxide nanoparticles on the salt stress responses of Brassica napus L. Environ Pollut, 2016, 219: 28-36

[36] RossiL, ZhangW, MaXM. Cerium oxide nanoparticles alter the salt stress tolerance of Brassica napus L. by modifying the formation of root apoplastic barriers. Environ Pollut, 2017, 229: 132-138

[37] Saad-AllahKM, RagabGA. Sulfur nanoparticles mediated improvement of salt tolerance in wheat relates to decreasing oxidative stress and regulating metabolic activity. Physiol Mol Biol Plants, 2020, 26(11):2209-2223

[38] Shabala S, Munns R (2017) Salinity stress: physiological constraints and adaptive mechanisms. In Shabala S (ed) Plant stress physiology, 2nd edn. CABI, Australia. https://doi.org/10.1079/9781780647296.0024

[39] ShrivastavaP, KumarR. Soil salinity: a serious environmental issue and plant growth promoting bacteria as one of the tools for its alleviation. Saudi J Biol Sci, 2015, 22(2):123-131

[40] SinghD, SilluD, KumarA, AgnihotriS. Dual nanozyme characteristics of iron oxide nanoparticles alleviate salinity stress and promote the growth of an agroforestry tree Eucalyptus Tereticornis Sm. Environ Sci Nano, 2021, 8(5):1308-1325

[41] SunJ, ChenS, DaiS, WangR, LiN, ShenX, ZhouX, LuC, ZhengX, HuZ, ZhangZ, SongJ, XuY. NaCl-induced alternations of cellular and tissue ion fluxes in roots of salt-resistant and salt-sensitive poplar species. Plant Physiol, 2009, 149(2):1141-1153

[42] van IttersumMK, van BusselLGJ, WolfJ, GrassiniP, van WartJ, GuilpartN, ClaessensL, de GrootH, WiebeK, Mason-D'CrozD, YangH, BoogaardH, van OortPAJ, van LoonMP, SaitoK, AdimoO, Adjei-NsiahS, AgaliA, BalaA, ChikowoR, KaizziK, KouressyM, MakoiJHJR, OuattaraK, TesfayeK, CassmanKG. Can sub-Saharan Africa feed itself?. PNAS, 2016, 113(52):14964-14969

[43] WahidI, RaniP, KumariS, AhmadR, HussainSJ, AlamriS, TripathyN, KhanMIR. Biosynthesized gold nanoparticles maintained nitrogen metabolism, nitric oxide synthesis, ions balance, and stabilizes the defense systems to improve salt stress tolerance in wheat. Chemosphere, 2022, 287(2

[44] WhatmoreAM, ReedRH. Determination of turgor pressure in Bacillus subtilis: a possible role for K⁺ in turgor regulation. J Gen Microbiol, 1990, 136 12):2521-2526

[45] WuHH, LiZH. Nano-enabled agriculture: how nanoparticles cross barriers in plants?. Plant Communications in Press, 2022

[46] WuHH, LiZH. Recent advances in nano-enabled agriculture for improving plant performance. Crop J, 2022, 10(1):1-12

[47] WuHH, ShabalaL, BarryK, ZhouM, ShabalaS. Ability of leaf mesophyll to retain potassium correlates with salinity tolerance in wheat and barley. Physiol Plant, 2013, 149(4):515-527

[48] WuHH, ShabalaL, ZhouM, ShabalaS. Durum and bread wheat differ in their ability to retain potassium in leaf mesophyll: implications for salinity stress tolerance. Plant Cell Physiol, 2014, 55(10):1749-1762

[49] WuHH, ShabalaL, ZhouM, ShabalaS. Chloroplast-generated ROS dominate NaCl^- induced K⁺ efflux in wheat leaf mesophyll. Plant Signal Behav, 2015, 10(5

[50] WuHH, TitoN, GiraldoJP. Anionic cerium oxide nanoparticles protect plant photosynthesis from abiotic stress by scavenging reactive oxygen species. ACS Nano, 2017, 11 11):11283-11297

[51] Wu HH, Shabala L, Shabala S, Giraldo JP (2018a) Hydroxyl radical scavenging by cerium oxide nanoparticles improves Arabidopsis salinity tolerance by enhancing leaf mesophyll potassium retention. Environ Sci Nano 5(7):1567–1583. https://doi.org/10.1039/c8en00323h

[52] Wu HH, Zhang XC, Giraldo JP, Shabala S (2018b) It is not all about sodium: revealing tissue specificity and signalling roles of potassium in plant responses to salt stress. Plant Soil 431(1–2):1–17. https://doi.org/10.1007/s11104-018-3770-y

[53] WuHH, ShabalaL, ZhouM, SuN, WuQ, Ul-HaqT, ZhuJ, MancusoS, AzzarelloE, ShabalaS. Root vacuolar Na⁺ sequestration but not exclusion from uptake correlates with barley salt tolerance. Plant J, 2019, 100(1):55-67

[54] YlvisakerJR. Required MD coverage last resort to solving hospitals' liability problems. Mich Med, 1982, 81(29):328-331

[55] YouJ, ChanZ. ROS regulation during abiotic stress responses in crop plants. Front Plant Sci, 2015, 6: 1092

[56] ZhaoL, LuL, WangA, ZhangH, HuangM, WuH, XingB, WangZ, JiR. Nano-biotechnology in agriculture: use of nanomaterials to promote plant growth and stress tolerance. J Agric Food Chem, 2020, 68(7):1935-1947

[57] ZhouH, WuHH, ZhangF, SuY, GuanWX, XieYJ, GiraldoJP, ShenWB. Molecular basis of cerium oxide nanoparticle enhancement of rice salt tolerance and yield. Environ Sci Nano, 2021, 8(11):3294-3311

[58] ZhuJK. Abiotic stress signaling and responses in plants. Cell, 2016, 167(2):313-324

Funding

National Natural Science Foundation of China(31901464); Northwest A&F University