Document Type : Original Article
Authors
1 Professor, Department of Plant Production and Genetics, Faculty of Agriculture and Natural Resources, University of Mohaghegh Ardabili, Ardabil, Iran
2 Ph.D, Plant Physiology, Department of Plant Production and Genetics, Faculty of Agriculture and Natural Resources, University of Mohaghegh Ardabili, Ardabil, Iran.
Abstract
One of the most critical limiting factors for cowpea seed germination (Vigna unguiculata subsp. unguiculata) is drought stress. This study aimed to examine the effect of seed priming with titanium dioxide (TiO₂) nanoparticles (NPs) under drought stress conditions, using a factorial experiment in a completely randomized design with three replications. Treatments included seed priming with TiO₂ NPs at concentrations of 0, 25, and 50 mg L⁻¹ and drought stress levels at 0, -0.025, -0.050, -0.075, and -0.1 MPa induced by polyethylene glycol 6000. Results showed broad absorption peaks in the ranges of 800–500 cm⁻¹ and 3600–3200 cm⁻¹, corresponding to Ti-O-Ti and O-H stretching vibrations, respectively. The TiO₂ NPs exhibited a spherical shape. Increasing drought stress significantly decreased germination percentage and both length and weight indices of the seedlings. The lowest germination percentage was recorded at -0.1 MPa osmotic potential, with a 83% reduction compared to the control. Seed priming with TiO₂ NPs at 25 and 50 mg L-1 did not significantly affect germination; however, the 50 mg L-1 concentration had a negative impact on seed germination traits. Seed priming at 25 mg L⁻¹ increased seedling plumule length by 15%. Additionally, the seed vigor index increased at this concentration across all drought stress levels. Catalase enzyme activity and proline content at (-0.1 MPa) osmotic potential increased by 18% and 56%, respectively, compared to the control. These results demonstrate a concentration-dependent effect of TiO₂ NPs in enhancing germination and the length and weight indices of cowpea seedlings.
Keywords
Abbasi Khalaki, M., Moameri, M., Asgari Lajayer, B., & Astatkie, T. (2021). Influence of nano-priming on seed germination and plant growth of forage and medicinal plants. Plant growth regulation, 93(1), 13-28. https://doi.org/10.1007/s10725-020-00670-9
Acharya, P., Jayaprakasha, GK., Crosby, KM., Jifon, JL., & Patil BS. (2020). Nanoparticle mediated seed priming improves germination, growth, yield, and quality of watermelons (Citrullus lanatus) at multi-locations in Texas. Scientific Reports, 19, 10(1):5037, https://doi.org/10.1038/s41598-020-61696-7
Ahmadnia, F., & Ebadi, A. 2024. Evaluation of improving Sunn hemp (Crotalaria juncea) seed germination with titanium dioxide (TiO2) nanoparticles under salinity stress. Iranian Journal of Seed Science and Research, 11(1), 69-87. https://doi.org/10.22124/jms.2024.8039 [In Persian]
Ahmadnia, F., Ebadi, A., Alebrahim, M.T., Parmoon, G., Feizpoor, S., & Hashemi, M. (2025). The efficacy of Sunn Hemp (Crotalaria juncea) and Fe3O4 nanoparticles in controlling weed seed germination. Agronomy, 15, 795, 1-21. https://doi.org/10.3390/agronomy15040795
Arun, M.N., Bhanuprakash, K., Shankar, S., & Senthivel, T. (2017). Effects of seed priming on biochemical parameters and seed germination in cowpea [Vigna unguiculata (L.) Walp]. Legume Research, 40(3), 562-570. https://doi.org/10.18805/lr.v0i0.7857
Asli, S., & Neumann, P.M. (2009). Colloidal suspensions of clay or titanium dioxide nanoparticles can inhibit leaf growth and transpiration via physical effects on root water transport. Plant, Cell and Environment, 32(5), 577-584. https://www.ncbi.nlm.nih.gov/pubmed/19210640
Bates, L.S., Walderen, R.D., & Taere I.D. (1973). Rapid determination of free proline for water stress studies. Plant Soil, 39, 205-207. https://doi.org/10.1007/BF00018060
Carvalho, M., Castro, I., Moutinho-Pereira, J., Correia, C., Egea-Cortines, M., Matos, M., Rosa, E., Carnide, V., & Lino-Neto, T. (2019). Evaluating stress responses in cowpea under drought stress. Journal of Plant Physiology, 241, 153001. https://doi.org/10.1016/j. jplph.2019.153001
Chandrasekaran, U., Luo, X., Wang, Q., & Shu, K. (2020). Are there unidentified factors involved in the germination of nano-primed seeds? Frontiers in Plant Science, 11, 832. https://doi.org/10.3389/fpls.2020.00832
Cook, B.I., Smerdon, J.E., Seager, R., & Coats, S. (2014). Global warming and 21st century drying. Climate Dynamics, 43, 2607–2627.
Cramer, G.R., Urano, K., Delrot, S., Pezzotti, M., & Shinozaki, K. (2011). Effects of abiotic stress on plants: a systems biology perspective. BMC Plant Biology, 11, 163. https://doi.org/10.1186/1471-2229-11-163
da Silva, A.C., da Costa Santos, D., Junior, D.L.T., da Silva, P.B., dos Santos, R.C., & Siviero, A. (2018). Cowpea: A Strategic Legume Species for Food Security and Health. In: Legume Seed Nutraceutical Research. IntechOpen. Chapter 3, 1-21. https://doi.org/10.5772/intechopen.79006
Ebadi, A., Feizpoor, S., & Ahmadnia, F. (2025). Investigating the impact of sunflower (Helianthus annuus) allelopathic aqueous extract in the green synthesis by titanium dioxide nanoparticles on wild mustard (Sinapis arvensis L.) seed germination. Plant Process and Function, 14 (66): 47-63. https://doi.org/10.22034/14.66.47
Fabunmi, T. O., Gbadamosi, B. K., & Adigbo, S. O. (2012). Seed hydro-priming and early moisture stress impact on biomass production and grain yield of cowpea. International Journal of Applied, 2(10), 112-122. https://www.researchgate.net/publication/272795669_Seed_Hydro-Priming_and_Early_Moisture_Stress_Impact_on_Biomass_Production_and_Grain_Yield_of_Cowpea
FAOSTAT, (2020). FAOSTAT: food and agriculture data. Available at: http://www.fao. org/ faostat/en/#home, Accessed date: 4 March 2021
Gerland, P., Raftery, A.E., Sev ˇ ˇcíkova, ´ H., Li, N., Gu, D., Spoorenberg, T., Alkema, L., Fosdick, B.K., Chunn, J., Lalic, N., Bay, G., Buettner, T., Heilig, G.K., & Wilmoth, J. (2014). World population stabilization unlikely this century. Science, 346, 234-237. https://doi.org/10.1126/science.1257469
Ghorbani, R., Movafeghi, A., Ganjeali, A., & Nabati, J. (2022). Investigating the germination characteristics of Chickpea (Cicer arietinum) in response to titanium dioxide nanoparticles priming and drought stress. Iranian Journal of Seed Research, 9(1), 189-202. https://doi.org/10.52547/yujs.9.1.189 [In Persian]
Goufo, P., Moutinho-Pereira, J.M., Jorge, T.F., Correia, C.M., Oliveira, M.R., Rosa, E.A.S., Antonio,´C., & Trindade, H. (2017). Cowpea (Vigna unguiculata L. Walp.) Metabolomics: Osmo protection as a physiological strategy for drought stress resistance and improved yield. Frontiers in Plant Science, 8:586, https://doi.org/10.3389/ fpls.2017.00586
Imtiaz, H., Shiraz, M., Mir, A.R., Siddiqui, H., & Hayat, A. (2023). Nano‑Priming Techniques for Plant Physio‑Biochemistry and Stress Tolerance. Journal of Plant Growth Regulation, 42(1), 1-21.https://doi.org/10.1007/s00344-023-10981-6
Kandhol, N., Singh, V. P., Ramawat, N., Prasad, R., Chauhan, D. K., Sharma, S., Grillo, R., Shai, S., Peralta-Videa, J., & Tripathi, D. K. (2022). Nano-priming: Impression on the beginner of plant life. Plant Stress, 5, 100091, 1-8. https://doi.org/10.1016/j.stress.2022.100091
Khalifa, A.M., Safhi, F.A., & Elsherif, D.E. (2024). Green synthesis of a dual-functional sulfur nanofertilizer to promote growth and enhance salt stress resilience in faba bean. BMC Plant Biology, 24, 607, 1-16. https://doi.org/10.1186/s12870-024-05270-7
Khan, MN., AlSolami, MA., Basahi, RA., Siddiqui, MH., Al-Huqail, AA., Abbas, ZK., Siddiqui, ZH., Ali, HM., & Khan, F. (2020). Nitric oxide is involved in nano-titanium dioxide-induced activation of antioxidant defense system and accumulation of osmolytes under water-deficit stress in Vicia faba L. Ecotoxicology Environmental Safety, 190,110152. https://doi.org/10.1016/j.ecoenv.2019.110152
Kobayashi, K., Kubota, H., Hojo, R., & Miyagawa, M. (2019). Effective dispersal of titanium dioxide nanoparticles for toxicity testing. Journal of Toxicological Sciences, 44(8), 515-521. https://doi.org/10.2131/jts.44.515
Ma, X., Geiser-Lee, J., Deng, Y., & Kolmakov, A. (2010). Interactions between engineered nanoparticles (ENPs) and plants: phytotoxicity, uptake and accumulation. Science of the Total Environment, 408(16), 3053-3061. https://doi.org/10.1016/j.scitotenv.2010.03.031
Maguire, J. D. (1962). Speed of germination-aid in selection and evaluation for seedling emergence and vigor. Crop Science., 2, 176-177. https://doi.org/10.2135/cropsci1962.0011183X000200020033x
Mahmoodzadeh, H., & Aghili, R. (2018). Effect on germination and early growth characteristics in wheat plants (Triticum aestivum L.) Seeds Exposed to TiO2 Nanoparticles. Journal of Chemical Health Risks, 4(1), 29-36.
Mathew, S.S., Sunny, N.E., & Shanmugam, V. (2021). Green synthesis of anatase titanium dioxide nanoparticles using Cuminum cyminum seed extract; effect on Mung bean (Vigna radiata) seed germination. Inorganic Chemistry Communications, 126, 108485, 1-18. https://doi.org/10.1016/j.inoche.2021.108485
Michel, B.E., & Kaufmann, M.R. (1973). The osmotic potential of polyethylene glycol 6000. Plant Physiology, 51(5), 914-916. https://doi.org/10.1104/pp.51.5.914
Mohammadi, H., Esmailpour, M., & Gheranpaye, A. (2016). Effects of TiO2 nanoparticles and water-deficit stress on morpho-physiological characteristics of dragonhead (Dracocephalum moldavica L.) plants. Acta Agriculturae Slovenica, 107(2), 385-396. https://doi.org/10.14720/aas.2016.107.2.11
Nadeem, M., Li, J., Yahya, M., Sher, A., Ma, C., Wang, X., & Qiu, L. (2019). Research progress and perspective on drought stress in legumes: a review. International Journal of Molecular Science, 23; 20(10): 2541. https://doi.org/10.3390/ijms20102541
Nil, S.H., Thiruvengadam, M., Wang, Y., Samynathan, R., Shariati, M.A., Rebezov, M., Nile, A., Sun, M., Venkidasamy, B., Xiao, J. & Kai, G. (2022). Nano-priming as emerging seed priming technology for sustainable agriculture—recent developments and future perspectives. Journal of Nanobiotechnology, (2022)20, 254, 1-31. https://doi.org/10.1186/s12951-022-01423-8
Noman, M.T., Ashraf, M.A., & Ali, A. (2019). Synthesis and applications of nano-TiO2: A review. Environmental Science and Pollution Research, 26, 3262-3291. https://doi.org/10.1007/s11356-018-3884-z
Olorunwa, O. J., Shi, A., & Barickman, T.C. (2021). Varying drought stress induces morpho-physiological changes in cowpea (Vigna unguiculata (L.) genotypes inoculated with Bradyrhizobium japonicum. Plant stress, 2(2021), 100033, 1-13. https://doi.org/10.1016/j.stress.2021.100033
Orchard, T. (1977). Estimating the parameters of plant seedling emergence. Seed Science and Technology, 5, 61-69.
Osakabe, Y., Osakabe, K., Shinozaki, K., Tran, L.S.P. (2014). Response of plants to water stress. Frontiers Plant Science. 5, 86. https://doi.org/10.3389/fpls.2014.00086
Paiva, E. P. D., Sá, F. V. D. S., Torres, S. B., Brito, M. E., Moreira, R. C., & Silva, L. D. A. (2018). Germination and tolerance of cowpea (Vigna unguiculata) cultivars to water stress. Revista Brasileira de Engenharia Agrícola e Ambiental, 22(6), 407-411. https://doi.org/10.1590/1807-929/agriambi.v22n6p407-411
Paiva, E. P. D., Sá, F. V. D. S., Torres, S. B., Brito, M. E., Moreira, R. C., & Silva, L. D. A. (2018). Germination and tolerance of cowpea (Vigna unguiculata) cultivars to water stress. Revista Brasileira de Engenharia Agrícola e Ambiental, 22(6), 407-411. https://doi.org/10.1590/1807-1929/agriambi.v22n6p407-411
Paramo, LA., Feregrino-Pe´rez, AA., Guevara, R., Mendoza, S., & Esquivel, K. (2020). Nanoparticles in agroindustry: applications, toxicity, challenges, and trends. Nanomaterials, 10, 1654. https://doi.org/10.3390/nano10091654
Pazhouhan, I., Jalali, S.Gh.A., Atabati, H., Zarafshar, M., & Sattarian, A. (2016). Comparison of carbon nanotubes with chemical and physical treatments to break seed dormancy of Myrtus communis L. Journal of Botany Research, 29(2), 300-308. [In Persian]
Perry, D.A. (1991). Methodology and application of vigor tests. International Seed Testing Association. Zurich. Switzerland.
Pham, X.N., Pham, D.T., Ngo, H.S., Nguyen, M.B., & Doan, H.V. (2021). Characterization and application of C–TiO2 doped cellulose acetate nanocomposite film for removal of Reactive Red-195. Chemical Engineering Communications, 208(3), 304-317. https://doi.org/10.1080/00986445.2020.1712375
Raliya, R., Nair, R., Chavalmane, S., Wang, W.N., & Biswas, P. (2015). Mechanistic evaluation of translocation and physiological impact of titanium dioxide and zinc oxide nanoparticles on the tomato (Solanum lycopersicum L.) plant. Metallomics, 7(12), 1584-1594. https://doi.org/10.1039/c5mt00168d
Ramadan, T., Sayed, S.A., Abd-Elaal, A. K.A., & Amro, A. (2022). The combined effect of water deficit stress and TiO2 nanoparticles on cell membrane and antioxidant enzymes in Helianthus annuus L. Physiology and Molecular Biology of Plants, 28(2), 391-409. https://doi.org/10.1007/s12298-022-01153-z
Rastogi, A., Zivcak, M., Sytar, O., Kalaji, HM., He, X., Mbarki, S., & Brestic, M. (2017). Impact of metal and metal oxide nanoparticles on plant: a critical review. Frontiers in Chemistry, 78:1-16. https://doi.org/10.3389/ fchem.2017.00078
Ravelombola, W., Shi, A., Chen, S., Xiong, H., Yang, Y., Cui, Q., Olaoye, D., & Mou, B. (2020). Evaluation of cowpea for drought tolerance at seedling stage. Euphytica, 216, 123. https://doi.org/10.1007/s10681-02 0-02660-4
Ravelombola, W., Shi, A., Qin, J., Weng, Y., Bhattarai, G., Zia, B., Zhou, W., & Mou, B. (2018). Investigation on various aboveground traits to identify drought tolerance in cowpea seedlings. HortScience, 53, 1757–1765
Rico, CM., Peralta-Videa, JR., & Gardea-Torresdey, JL. (2015). Chemistry, biochemistry of nanoparticles, and their role in antioxidant defense system in plants. Nanotechnology and Plant Science, 2:1–17. https://doi.org/10.1007/978-3-319-14502-0_1
Saeb, H., Khayyat, M., Zarezadeh, A., Moradinezhad, F., Samadzadeh, A., & Safaee, M. (2013). Effects of NaCl stress on seed germination attributes of periwinkle (Catharanthus roseus L.) and corn poppy (Papaver rhoeas L.) plants. Plant Breeding and Seed Science, 67,115-123
Sant´as-Miguela, V., Arias-Estevez, M., Rodriuez-Seijo, A., & Arenas-Lago, D. (2023). Use of metal nanoparticles in agriculture. A review on the effects on plant germination. Environmental Pollution, 334(2023), 122222, 1-17. https://doi.org/10.1016/j.envpol.2023.122222
Sauret-Güeto, S., Calder, G., & Harberd, N.P. (2012). Transient gibberellin application promotes Arabidopsis thaliana hypocotyl cell elongation without maintaining transverse orientation of microtubules on the outer tangential wall of epidermal cells. Plant Journal, 69(4), 628-639. https://doi.org/10.1111/j.1365-313X.2011.04817.x
Scott, S., Jones, R., & Williams, W. (1984). Review of data analysis methods for seed germination 1. Crop Science, 24(6), 1192-1199. https://doi.org/10.2135/cropsci1984.0011183X002400060043x
Servin, A., Elmer, W., Mukherjee, A., De la Torre-Roche, R., Hamdi, H., White, J.C., Bindraban, P., & Dimkpa, C.A. (2015). Review of the use of engineered nanomaterials to suppress plant disease and enhance crop yield. Journal of Nanoparticle Research, 17(2): 1-21. https://doi.org/10.1007/s11051-015-2907-7
Sharif, H., Mehmood, A., Ulfat, A., Ahmad, K.S., Hussain, I., & Khan, R.T. (2021). Environmentally sustainable production of silver nanoparticles and their effect on Glycine max L. Seedlings. Gesunde Pflanzen, 73(2021), 95-103. https://doi.org/10.1007/ s10343-020-00532-4
Singh, B. (2020). Cowpea: The Food Legume of the 21st Century. John Wiley & Sons.
Stewart, R.C., & Beweley, J.D. (1980). Lipid peroxidation associated with accelerated aging of soybean axes. Plant Physiology, 65(2), 245248. https://doi.org/10.1104/pp.65.2.245
Sudhakar, C., Lakshmi, A., & Giridara Kumar, S. (2001). Changes in the antioxidant enzyme efficacy in two high yielding genotypes of mulberry (Morus alba L.) under NaCl salinity. Plant Science, 167(3), 613-619. https://doi.org/10.1016/S0168-9452(01)00450-2
Tan, W., Peralta-Videa, JR., & Gardea-Torresdey, JL. (2018). Interaction of titanium dioxide nanoparticles with soil components and plants: current knowledge and future research needs–a critical review. Environmental Science: Nano Journal 5(8), 257–278. https://doi.org/10.1039/ C7EN00985B
Tekrony, D.M., & Egli, D.B. (1991). Relationship of seed vigor to crop yield: a review. Crop science, 31(3), 816-822. https://doi.org/10.2135/cropsci1991.0011183X003100030054x
Torre-Roche, RDL., Cantu, J., Tamez, C., Zuverza-Mena, N., Hamdi, H., Adisa, IO., Elmer, W., Gardea-Torresdey, J., & White, JC. (2020). Seed biofortification by engineered nanomaterials: A pathway to alleviate malnutrition? Journal of Agricultural and Food Chemistry, 68(44), 12189-202. https://doi.org/10.1021/acs.jafc.0c048 81
Vashisth, A., & Nagarajan, S. (2010). Effect on germination and early growth characteristics in sunflower (Helianthus annuus) seeds exposed to static magnetic field. Journal of Plant Physiology, 167(2), 149-156.
Wahid, I., Kumari, S., Ahmad, R., Hussain, SJ., Alamri, S., Siddiqui, MH., & Khan, MIR. (2020). Silver nanoparticle regulates salt tolerance in wheat through changes in ABA concentration, ion homeostasis, and defense systems. Biomolecules, 10(11), 1506, 1-19. https://doi.org/10. 3390/biom10111506
Waqas, MA., Kaya, C., Riaz, A., Farooq, M., Nawaz, I., Wilkes, A., & Li, Y. (2019). Potential mechanisms of abiotic stress tolerance in crop plants induced by Thiourea. Frontiers in Plant Science, 10, 1336. https://doi.org/10.3389/fpls.2019. 01336
Yuan, X., Wang, X., Zhang, Sh., Dong, X., Zhang, Sh., Zhou, W., Zhang, Ch., & Luo, Y. (2021). Synergistic enhancement of photocatalytic performance of mesoporous TiO2 enabled by tunable crystal phase and hybridization with graphene oxide. Chemistry Select, 6(23), 5791-5800. https://doi.org/10.1002/slct.202100784
)