Abstract
Nitrogen (N) is an essential nutrient for plants. Nitrate is often the main inorganic N form absorbed, and amino acids are the predominant organic N form transported in most plant species. Amino acid transporters (AATs) mediate the distribution of amino acids between organs, and any disturbance in this process can cause changes in crop development. To date, the OsAAP1 amino acid transporter has been characterized in the most detail during the reproductive and ripening periods in rice plants. In this study, we identified the sites of OsAAP1 expression during the early stages of rice development and determined the implications of OsAAP1 knockout on seedling establishment, nitrate–N uptake and assimilation, and the expression of genes related to N and carbon (C) metabolism. OsAAP1 is strongly expressed in the coleorhiza, radicle, and epicotyl of seedlings and in the regions of the vascular bundles of roots and leaves. OsAAP1 knockout hindered rice seedling establishment and impaired nitrate absorption. Under low N, OsAAP1 knockout results in damage to NO3− absorption, with negative effects on N assimilation and C metabolism. Under sufficient N, nitrate absorption was more strongly impaired, and downregulation of nitrate transporters and N assimilation enzymes was observed. Taken together, the data suggest that the OsAAP1 gene is essential for the early development of rice when nitrate is the predominant N source.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data Availability
The datasets generated during current study are available from the corresponding author on reasonable request.
References
Amir R, Galili G, Cohen H (2018) The metabolic roles of free amino acids during seed development. Plant Sci 275:11–18
Angelovici R, Fait A, Fernie AR, Galili G (2011) A seed high-lysine trait is negatively associated with the TCA cycle and slows down Arabidopsis seed germination. New Phytol 189:148–159. https://doi.org/10.1111/j.1469-8137.2010.03478.x
Arruda LN, Bucher CA, Rangel RP et al (2018) Overexpression of nitrate transporter OsNPF4.11 (OsNRT1.2) Affects growth in rice plants. Revista Brasileira De Ciencias Agrarias 13:1–8. https://doi.org/10.5039/agraria.v13i1a5493
Baslam M, Mitsui T, Sueyoshi K, Ohyama T (2021) Recent advances in carbon and nitrogen metabolism in C3 plants. Int J Mol Sci 22:1–39. https://doi.org/10.3390/ijms22010318
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)90527-3
Dechorgnat J, Nguyen CT, Armengaud P et al (2011) From the soil to the seeds: The long journey of nitrate in plants. J Exp Bot 62:1349–1359
Fagodiya RK, Pathak H, Bhatia A et al (2020) Global warming impacts of nitrogen use in agriculture: an assessment for India since 1960. Carbon Manag 11:291–301. https://doi.org/10.1080/17583004.2020.1752061
Fang Z, Wu B, Ji Y (2021) The amino acid transporter OsAAP4 contributes to rice tillering and grain yield by regulating neutral amino acid allocation through two splicing variants. Rice. https://doi.org/10.1186/s12284-020-00446-9
Farnden KJS, Robertson JG (1980) Methods for studying enzyme involved in metabolism related to nitrogen. In: Bergsen FJ (Ed.), Methods for Evaluating Biological Nitrogen Fixation. J W S 265–314
Felker P (1977) Microdetermination of nitrogen in seed protein extracts with the salicylate-dichloroisocyanurate color reaction. Anal Chem 49(7):1080–1080
Fernandes MS (1984) N-carriers, light and temp influences on free amino pool composition of rice plants. Turrialba. 297–301
Ferreira LM, da Rocha JG, Santos LA et al (2015) Cinética de absorção e frações nitrogenadas em arroz expressando o fator de transcrição OsDof26. Revista Ciencia Agronomica 46:379–387. https://doi.org/10.5935/1806-6690.20150017
Forde BG (2014) Glutamate signalling in roots. J Exp Bot 65:779–787
Gao J, Liu J, Li B, Li Z (2001) Isolation and purification of functional total RNA from blue-grained wheat endosperm tissues containing high levels of starches and flavonoids. Plant Mol Biol Report 19:185–186. https://doi.org/10.1007/BF02772163
Gent L, Forde BG (2017) How do plants sense their nitrogen status? J Exp Bot 68:2531–2540
Guo N, Gu M, Hu J et al (2020) Rice OsLHT1 functions in leaf-to-panicle nitrogen allocation for grain yield and quality. Front Plant Sci 11:1–13. https://doi.org/10.3389/fpls.2020.01150
Guo N, Zhang S, Gu M, Xu G (2021) Function, transport, and regulation of amino acids: What is missing in rice? Crop Journal 9:530–542. https://doi.org/10.1016/j.cj.2021.04.002
Han C, Yang P (2015) Studies on the molecular mechanisms of seed germination. Proteomics 15:1671–1679
Heinemann B, Hildebrandt TM (2021) The role of amino acid metabolism in signaling and metabolic adaptation to stress-induced energy deficiency in plants. J Exp Bot 72:4634–4645. https://doi.org/10.1093/jxb/erab182
Jefferson RA, Kavanagh TA, Bevan MW (1987) GUS fusions: beta-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 6:3901–3907. https://doi.org/10.1002/j.1460-2075.1987.tb02730.x
Ji Y, Huang W, Wu B et al (2020) The amino acid transporter AAP1 mediates growth and grain yield by regulating neutral amino acid uptake and reallocation in Oryza sativa. J Exp Bot 71:4763–4777. https://doi.org/10.1093/jxb/eraa256
Khaliq A, Aslam F, Matloob A et al (2015) Seed priming with selenium: Consequences for emergence, seedling growth, and biochemical attributes of rice. Biol Trace Elem Res 166:236–244. https://doi.org/10.1007/s12011-015-0260-4
Kihara T, Ohno T, Koyama H, et al (2003) Characterization of NADP-isocitrate dehydrogenase expression in a carrot mutant cell line with enhanced citrate excretion
Léran S, Varala K, Boyer JC et al (2014) A unified nomenclature of nitrate transporter 1/peptide transporter family members in plants. Trends Plant Sci 19:5–9
Li SX, Wang ZH, Stewart BA (2013) Responses of Crop Plants to Ammonium and Nitrate N. In: Advances in Agronomy. Academic Press Inc., pp 205–397
Liao HS, Chung YH, Hsieh MH (2022) Glutamate: a multifunctional amino acid in plants. Plant Sci. https://doi.org/10.1016/j.plantsci.2022.111238
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method. Methods 25:402–408. https://doi.org/10.1006/meth.2001.1262
Lu K, Wu B, Wang J et al (2018) Blocking amino acid transporter OsAAP3 improves grain yield by promoting outgrowth buds and increasing tiller number in rice. Plant Biotechnol J 16:1710–1722. https://doi.org/10.1111/pbi.12907
Miller AJ, Fan X, Shen Q, Smith SJ (2008) Amino acids and nitrate as signals for the regulation of nitrogen acquisition. In: Journal of Experimental Botany. pp 111–119
Miranda KM, Espey MG, Wink DA (2001) A rapid, simple spectrophotometric method for simultaneous detection of nitrate and nitrite. Nitric Oxide 5:62–71. https://doi.org/10.1006/niox.2000.0319
Muller B, Touraine B (1992) Inhibition of NO3" uptake by various phloem-translocated amino acids in soybean seedlings. J Exp Bot. https://doi.org/10.1093/jxb/43.5.617
Nan N, Wang J, Shi Y et al (2020) Rice plastidial NAD-dependent malate dehydrogenase 1 negatively regulates salt stress response by reducing the vitamin B6 content. Plant Biotechnol J 18:172–184. https://doi.org/10.1111/pbi.13184
Nazish T, Arshad M, Jan SU et al (2022) Transporters and transcription factors gene families involved in improving nitrogen use efficiency (NUE) and assimilation in rice (Oryza sativa L.). Transgenic Res 31:23–42
Ogasawara S, Ezaki M, Ishida R et al (2021) Rice amino acid transporter-like 6 (OsATL6) is involved in amino acid homeostasis by modulating the vacuolar storage of glutamine in roots. Plant J 107:1616–1630. https://doi.org/10.1111/tpj.15403
Peng B, Kong H, Li Y et al (2014) OsAAP6 functions as an important regulator of grain protein content and nutritional quality in rice. Nat Commun 5:1–12. https://doi.org/10.1038/ncomms5847
Pereira EG, Bucher CPC, Bucher CA et al (2022) The amino acid transporter OsAAP1 regulates the fertility of spikelets and the efficient use of N in rice. Plant Soil. https://doi.org/10.1007/s11104-022-05598-9
Qiu XM, Sun YY, Ye XY, Li ZG (2020) Signaling role of glutamate in plants. Front Plant Sci. https://doi.org/10.3389/fpls.2019.01743
Santos KFD, Eça N, Silveira RDD, Martin-Didonet CCG, Brondani C (2013) Storage protein profile and amino acid content in wild rice oryza glumaepatula. Pesqui Agropecu Bras 48:66–72. https://doi.org/10.1590/S0100-204X2013000100009
Santos LA, Santos WA, Sperandio MVL et al (2011) Nitrate uptake kinetics and metabolic parameters in two rice varieties grown in high and low nitrate. J Plant Nutr 34(7):988–1002. https://doi.org/10.1080/01904167.2011.555581
Shen T, Xiong Q, Zhong L et al (2019) Analysis of main metabolisms during nitrogen deficiency and compensation in rice. Acta Physiol Plant 41:1–14. https://doi.org/10.1007/s11738-019-2860-7
Tegeder M (2014) Transporters involved in source to sink partitioning of amino acids and ureides: opportunities for crop improvement. J Exp Bot 65:1865–1878. https://doi.org/10.1093/jxb/eru012
Tegeder M, Masclaux-Daubresse C (2018) Source and sink mechanisms of nitrogen transport and use. New Phytol 217:35–53. https://doi.org/10.1111/nph.14876
Wang B, Zhou G, Guo S et al (2022) Improving nitrogen use efficiency in rice for sustainable agriculture: strategies and future perspectives. Life. https://doi.org/10.3390/life12101653
Wang J, Wu B, Lu K et al (2019a) The amino acid permease 5 (Osaap5) regulates tiller number and grain yield in rice. Plant Physiol 180:1031–1045. https://doi.org/10.1104/pp.19.00034
Wang W, Hu B, Li A, Chu C (2020) NRT1.1s in plants: Functions beyond nitrate transport. J Exp Bot 71:4373–4379. https://doi.org/10.1093/jxb/erz554
Wang X, Yang G, Shi M et al (2019b) Disruption of an amino acid transporter LHT1 leads to growth inhibition and low yields in rice. BMC Plant Biol 19:1–11. https://doi.org/10.1186/s12870-019-1885-9
Xiong Q, Tang G, Zhong L et al (2018) Response to nitrogen deficiency and compensation on physiological characteristics, yield formation, and nitrogen utilization of rice. Front Plant Sci 9:1–14. https://doi.org/10.3389/fpls.2018.01075
Yang M, Yang J, Su L et al (2019) Metabolic profile analysis and identification of key metabolites during rice seed germination under low-temperature stress. Plant Sci. https://doi.org/10.1016/j.plantsci.2019.110282
Yang X, Yang G, Wei X et al (2023) OsAAP15, an amino acid transporter in response to nitrogen concentration, mediates panicle branching and grain yield in rice. Plant Sci 330:111640. https://doi.org/10.1016/j.plantsci.2023.111640
Yemm EI, Cocking ECC (1955) The determination of amino-acids with ninhydrin. Analyst. https://doi.org/10.1039/an9558000209
Yemm EW, Willis AJ (1954) The Estimation of Carbohydrates in Plant Extracts by Anthrone. Cambridge University Press
Zhang Y, Xiao W, Luo L et al (2012) Downregulation of OsPK1, a cytosolic pyruvate kinase, by T-DNA insertion causes dwarfism and panicle enclosure in rice. Planta 235:25–38. https://doi.org/10.1007/s00425-011-1471-3
Zhao H, Ma H, Yu L et al (2012) Genome-wide survey and expression analysis of amino acid transporter gene family in rice (Oryza sativa L). PLoS One. https://doi.org/10.1371/journal.pone.0049210
Funding
We thank the National Council for Scientific and Technological Development (CNPq), Rio de Janeiro Research Foundation (FAPERJ) and the Coordination for the Improvement of Higher-Brazil (CAPES) for granting fellowships for the authors. This work was financially supported by the Coordination for the Improvement of Higher-Brazil (CAPES)-Finance Code 001., Rio de Janeiro Research Foundation (FAPERJ), and the Plant Mineral Nutrition Laboratory (LNMP).
Author information
Authors and Affiliations
Contributions
EGP contributed toward conceptualization, experimental design, investigation, formal analysis, data analysis, and writing – original draft. LAS and MSF contributed toward conceptualization, writing – reviewing and editing, and supervision. ACOC, YRSR, CSC, CPCB, CAB, and ACG contributed toward investigation and formal analysis.
Corresponding author
Ethics declarations
Competing interest
The authors declare none.
Additional information
Handling Author: Rupesh Deshmukh.
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
Pereira, E.G., Santos, L.A., Chapeta, A.C.O. et al. Disruption of Amino Acid Transporter OsAAP1 Impairs Rice Seedling Establishment and Nitrate Uptake and Assimilation. J Plant Growth Regul (2024). https://doi.org/10.1007/s00344-024-11312-z
Received:
Accepted:
Published:
DOI: https://doi.org/10.1007/s00344-024-11312-z