biologia plantarum

International journal on Plant Life established by Bohumil Nìmec in 1959

Biologia plantarum 69:12-20, 2025 | DOI: 10.32615/bp.2025.002

Effects of NO3-/NH4+ ratios on growth, enzyme activity and nitrogen assimilation-related gene expression in Toona sinensis seedlings

Xiaopu SHI, Taotao SHAO, Beibei MA, Juan WANG, Mingqin FAN, Hu ZHAO*
Biology and Food Engineering College, Fuyang Normal University, Anhui 236037, People's Republic of China

Nitrogen is an essential nutrient for plants. Different nitrate (NO3-)/ammonium (NH4+) ratios have different effects on plant growth. However, the underlying mechanism in Toona sinensis remains unclear. Thus, we determined the effects of five different NO3-/NH4+ ratios (16/0, 12/4, 8/8, 4/12, and 0/16, denoted T1, T2, T3, T4, and T5, respectively) in nutrient media on T. sinensis seedling growth. When the nitrogen source was NH4+ alone (T5) or NO3- alone (T1), the soluble protein content in the leaves was the lowest. Additionally, the activities of key nitrogen assimilation-related enzymes, such as nitrate reductase (NR), glutamate synthase (GOGAT), and glutamine synthetase (GS), were altered by the NO3-/NH4+ ratio. Principal component analysis (PCA) revealed that the T2 treatment was optimal for T. sinensis seedling growth. The NO3-/NH4+ ratio regulates nitrogen assimilation at the transcription level, as under high NO3- conditions, the expressions of NR, GS, and NADH-GOGAT were high, and nitrate transporter (NRT) family members NRT1, NRT1.1, and NRT1.7 played leading roles in nitrogen transport. However, under low NO3- conditions, the level of NRT2.7 increased to ensure nutrient absorption. Our results provide a theoretical basis for understanding how different NO3-/NH4+ ratios affect T. sinensis growth.

Keywords: ammonium, ammonium transporter, Chinese toon, nitrate, nitrate transporter, nitrogen assimilation-related enzymes.

Received: September 4, 2024; Revised: January 31, 2025; Accepted: March 14, 2025; Published online: April 1, 2025  Show citation

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SHI, X., SHAO, T., Beibei, M., WANG, J., FAN, M., & ZHAO, H. (2025). Effects of NO3-/NH4+ ratios on growth, enzyme activity and nitrogen assimilation-related gene expression in Toona sinensis seedlings. Biologia plantarum, 69, Article 12-20. https://doi.org/10.32615/bp.2025.002
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    References

    1. Andrews, M., Raven, J.A. & Lea, P.J. (2013) Do plants need nitrate? The mechanisms by which nitrogen form affects plants. Annals of Applied Biology, 163, 174-199. Go to original source...
    2. Bernard, S.M. & Habash, D.Z. (2009) The importance of cytosolic glutamine synthetase in nitrogen assimilation and recycling. New Phytologist, 182, 608-620. Go to original source...
    3. Bradford, M.M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72, 248-254. Go to original source...
    4. Chen, H., Zhang, Q., Cai, H., Zhou, W. & Xu, F. (2018) H2O2 mediates nitrate-induced iron chlorosis by regulating iron homeostasis in rice. Plant, Cell and Environment, 41, 767-781. Go to original source...
    5. Cruz, C., Domínguez-Valdivia, M.D., Aparicio-Tejo, P.M. et al. (2011) Intra-specific variation in pea responses to ammonium nutrition leads to different degrees of tolerance. Environmental and Experimental Botany, 70, 233-243. Go to original source...
    6. Ferreira, S., Moreira, E., Amorim, I., Santos, C. & Melo, P. (2019) Arabidopsis thaliana mutants devoid of chloroplast glutamine synthetase (GS2) have non-lethal phenotype under photorespiratory conditions. Plant Physiology and Biochemistry, 144, 365-374. Go to original source...
    7. Fortunato, S., Nigro, D., Lasorella, C., Marcotuli, I., Gadaleta, A. & de Pinto, M.C. (2023) The role of glutamine synthetase (GS) and glutamate synthase (GOGAT) in the improvement of nitrogen use efficiency in cereals. Biomolecules, 13, 1771. Go to original source...
    8. Foyer, C.H., Noctor, G. & Hodges, M. (2011) Respiration and nitrogen assimilation: targeting mitochondria-associated metabolism as a means to enhance nitrogen use efficiency. Journal of Experimental Botany, 62, 1467-1482. Go to original source...
    9. Gao, Z, Wang, Y, Chen, G. et al. (2019) The indica nitrate reductase gene OsNR2 allele enhances rice yield potential and nitrogen use efficiency. Nature Communications, 10, 5207. Go to original source...
    10. Guo, S., Chen, G., Zhou, Y. & Shen, Q. (2007) Ammonium nutrition increases photosynthesis rate under water stress at early development stage of rice (Oryza sativa L.). Plant and Soil, 296, 115-124. Go to original source...
    11. Hachiya, T. & Sakakibara, H. (2017) Interactions between nitrate and ammonium in their uptake, allocation, assimilation, and signaling in plants. Journal of Experimental Botany, 68, 2501-2512. Go to original source...
    12. Hu, L., Yu, J., Liao, W. et al. (2015) Moderate ammonium:nitrate alleviates low light intensity stress in mini Chinese cabbage seedling by regulating root architecture and photosynthesis. Scientia Horticulturae, 186, 143-153. Go to original source...
    13. Huang, N.-C., Liu, K.-H., Lo, H.-J. & Tsay, Y.-F. (1999) Cloning and functional characterization of an Arabidopsis nitrate transporter gene that encodes a constitutive component of low-affinity uptake. The Plant Cell, 11, 1381-1392. Go to original source...
    14. Krapp, A., David, L.C., Chardin, C. et al. (2014) Nitrate transport and signalling in Arabidopsis. Journal of Experimental Botany, 65, 789-798. Go to original source...
    15. Kuppe, C.W., Johannes, A. & Postma, J.A. (2024) Benefits and limits of biological nitrification inhibitors for plant nitrogen uptake and the environment. Scientific Reports, 14, 15027. Go to original source...
    16. Lin, Y.M., Tay, J.H., Liu, Y. & Hung, Y.T. (2009) Biological nitrification and denitrification processes. In: Wang, L.K., Pereira, N.C. & Hung, Y.T. (Eds.) Biological Treatment Processes. Handbook of Environmental Engineering. Vol. 8. Totowa: Humana Press, pp. 539-588. Go to original source...
    17. Liu, W., Li, Y., Tomasetto, F. et al. (2022) Non-destructive measurements of Toona sinensis chlorophyll and nitrogen content under drought stress using near infrared spectroscopy. Frontiers in Plant Science, 12, 809828. Go to original source...
    18. Livak, K.J. & Schmittgen, T.D. (2001). Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method. Methods, 25, 402-408. Go to original source...
    19. Loqué, D. & von Wirén, N. (2004) Regulatory levels for the transport of ammonium in plant roots. Journal of Experimental Botany, 55, 1293-1305. Go to original source...
    20. Maathuis, F.J.M. (2009) Physiological functions of mineral macronutrients. Current Opinion in Plant Biology, 12, 250-258. Go to original source...
    21. Masclaux-Daubresse, C., Daniel-Vedele, F., Dechorgnat, J., Chardon, F., Gaufichon, L. & Suzuki, A. (2010) Nitrogen uptake, assimilation and remobilization in plants: challenges for sustainable and productive agriculture. Annals of Botany, 105, 1141-1157. Go to original source...
    22. Meyer, C. & Stitt, M. (2001) Nitrate reduction and signalling. In: Lea, P.J., Morot-Gaudry, J.-F. (Eds.) Plant Nitrogen. Berlin-Heidelberg: Springer, pp. 37-59. Go to original source...
    23. Miflin, B.J. & Habash, D.Z. (2002) The role of glutamine synthetase and glutamate dehydrogenase in nitrogen assimilation and possibilities for improvement in the nitrogen utilization of crops. Journal of Experimental Botany, 53, 979-987. Go to original source...
    24. Nacry, P., Bouguyon, E. & Gojon, A. (2013) Nitrogen acquisition by roots: physiological and developmental mechanisms ensuring plant adaptation to a fluctuating resource. Plant and Soil, 370, 1-29. Go to original source...
    25. Oldroyd, G.E.D. & Leyser, O. (2020) A plant's diet, surviving in a variable nutrient environment. Science, 368, eaba0196. Go to original source...
    26. Pratiwi, E.P.A., Hillary, A.K., Fukuda, T. & Shinogi, Y. (2016) The effects of rice husk char on ammonium, nitrate and phosphate retention and leaching in loamy soil. Geoderma, 277, 61-68. Go to original source...
    27. Praxedes, S.C., DaMatta, F.M., Loureiro, M.E., Ferrão, M.A.G. & Cordeiro, A.T. (2006) Effects of long-term soil drought on photosynthesis and carbohydrate metabolism in mature robusta coffee (Coffea canephora Pierre var. kouillou) leaves. Environmental and Experimental Botany, 56, 263-273. Go to original source...
    28. Sun, X., Chen, F., Yuan, L. & Mi, G. (2020) The physiological mechanism underlying root elongation in response to nitrogen deficiency in crop plants. Planta, 251, 84. Go to original source...
    29. Suzuki, A. & Knaff, D.B. (2005) Glutamate synthase: structural, mechanistic and regulatory properties, and role in the amino acid metabolism. Photosynthesis Research, 83, 191-217. Go to original source...
    30. Tabatabaei, S.J., Yusefi, M. & Hajiloo, J. (2008) Effects of shading and NO3:NH4 ratio on the yield, quality and N metabolism in strawberry. Scientia Horticulturae, 116, 264-272. Go to original source...
    31. Tabuchi, M., Abiko, T. & Yamaya, T. (2007) Assimilation of ammonium ions and reutilization of nitrogen in rice (Oryza sativa L.). Journal of Experimental Botany, 58, 2319-2327. Go to original source...
    32. Unno, H., Uchida, T., Sugawara, H. et al. (2006) Atomic structure of plant glutamine synthetase: a key enzyme for plant productivity. Journal of Biological Chemistry, 281, 29287-29296. Go to original source...
    33. Vanoni, M.A., Dossena, L., van den Heuvel, R.H.H. & Curti, B. (2005) Structure-function studies on the complex iron-sulfur flavoprotein glutamate synthase: the key enzyme of ammonia assimilation. Photosynthesis Research, 83, 219-238. Go to original source...
    34. Wang, R., Tischner, R., Gutiérrez, R.A. et al. (2004) Genomic analysis of the nitrate response using a nitrate reductase-null mutant of Arabidopsis. Plant Physiology, 136, 2512-2522. Go to original source...
    35. Wang, Y., Zhang, X., Liu, H., Sun, G., Song, S. & Chen, R. (2022) High NH4+/NO3- ratio inhibits the growth and nitrogen uptake of Chinese kale at the late growth stage by ammonia toxicity. Horticulturae 8, 8. Go to original source...
    36. Wang, Y.-Y., Cheng, Y.-H., Chen, K.-E. & Tsay, Y.-F. (2018) Nitrate transport, signaling, and use efficiency. Annual Review of Plant Biology, 69, 85-122. Go to original source...
    37. Williams, L.E. & Miller, A.J. (2001) Transporters responsible for the uptake and partitioning of nitrogenous solutes. Annual Review of Plant Biology, 52, 659-688. Go to original source...
    38. Witham, F.H., Blaydes, D.F. & Devlin, R.M. (1971) Experiments in Plant Physiology. New York: Van Nostrand Reinhold Co., pp. 167-200.
    39. Xing, J., Cao, X., Zhang, M., Wei, X., Zhang, J. & Wan, X. (2023) Plant nitrogen availability and crosstalk with phytohormones signallings and their biotechnology breeding application in crops. Plant Biotechnology Journal, 21, 1320-1342. Go to original source...
    40. Xuan, W., Beeckman, T. & Xu, G. (2017) Plant nitrogen nutrition: sensing and signaling. Current Opinion in Plant Biology, 39, 57-65. Go to original source...
    41. Yan, Y., Zhang, Z., Sun, H. et al. (2023) Nitrate confers rice adaptation to high ammonium by suppressing its uptake but promoting its assimilation. Molecular Plant, 16, 1871-1874. Go to original source...
    42. Yuan, L., Loqué, D., Kojima, S. et al. (2007) The organization of high-affinity ammonium uptake in Arabidopsis roots depends on the spatial arrangement and biochemical properties of AMT1-type transporters. The Plant Cell, 19, 2636-2652. Go to original source...
    43. Zhang, J., Lv, J., Dawuda, M.M. et al. (2019) Appropriate ammonium-nitrate ratio improves nutrient accumulation and fruit quality in pepper (Capsicum annuum L.). Agronomy, 9, 683. Go to original source...
    44. Zhang, Y.-Y., Tian, J.-P., Cui, J., Hong, Y.-H. & Luan, Y.-S. (2021) Effects of different NH4+/NO3- ratios on the photosynthetic and physiology responses of blueberry (Vaccinium spp.) seedlings growth. Journal of Plant Nutrition, 44, 854-864. Go to original source...
    45. Zhao, H., Ge, M.M., Zhang, F.Z. et al. (2024) Integrated morphological, physiological and transcriptomic analyses reveal the responses of Toona sinensis seedlings to low-nitrogen stress. Genomics, 116, 110899. Go to original source...
    46. Zhu, Y., Huang, X., Hao, Y. et al. (2020) Ammonium transporter (BcAMT1.2) mediates the interaction of ammonium and nitrate in Brassica campestris. Frontiers in Plant Science, 10, 1776. Go to original source...
    47. Zhu, Y., Li, G., Liu, H., Sun, G., Chen, R. & Song, S. (2018) Effects of partial replacement of nitrate with different nitrogen forms on the yield, quality and nitrate content of Chinese kale. Communications in Soil Science and Plant Analysis, 49, 1384-1393. Go to original source...
    48. Zhu, Y., Lian, J., Zeng, H. et al. (2011) Involvement of plasma membrane H+-ATPase in adaption of rice to ammonium nutrient. Rice Science, 18, 335-342. Go to original source...
    49. Zhu, Y., Qi, B., Hao, Y. et al. (2021) Appropriate NH4+/NO3- ratio triggers plant growth and nutrient uptake of flowering Chinese cabbage by optimizing the pH value of nutrient solution. Frontiers in Plant Science, 12, 656144. Go to original source...

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