Abstract
Aims
Amino acids are the main forms of organic nitrogen in the metabolism of higher plants. The translocation and allocation of these nutrients are performed by a series of amino acid transporters (AATs), which are essential for the efficient use of nitrogen and grain yield. The aims of this study were to analyze the presence of the OsAAP1 transporter in vegetative and reproductive tissues and to determine its role in nitrogen distribution and rice grain yield.
Methods
The promoter of OsAAP1 gene was fused to a GUS reporter and its activity was accessed in vegetative and reproductive organs/tissues. Using the CRISPR/Cas9 system we obtained aap1 mutant lines for study of agronomic traits, N use parameters, grain quality and nitrogen metabolism genes expression.
Results
OsAAP1 was located in the leaves, rachis, internode, seed coat, stamens, and embryos. Compared to the wild-type plants, the osaap1 mutant lines showed a reduction in growth and grain yield, as well as in the absorption, use, and remobilization of nitrogen. The distribution of amino acids was affected in the mutant lines, which resulted in the accumulation of amino acids in the vegetative organs. The fertility of the spikelets was severely reduced in the mutant lines. It was not possible to identify any compensatory effect on the loss of OsAAP1 function.
Conclusions
Together, these results demonstrate that the OsAAP1 transporter plays a critical role in the translocation of amino acids to reproductive organs and the maintenance of spikelet fertility and, consequently, grain yield.
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
Abiko T, Obara M, Ushioda A et al (2005) Localization of NAD-isocitrate dehydrogenase and glutamate dehydrogenase in rice roots: Candidates for providing carbon skeletons to NADH-glutamate synthase. Plant Cell Physiol 46:1724–1734. https://doi.org/10.1093/pcp/pci188
Bandumula N (2018) Rice production in Asia: key to global food security. Proc Natl Acad Sci India Sect B - Biol Sci 88:1323–1328. https://doi.org/10.1007/s40011-017-0867-7
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
Carcea M (2021) Value of wholegrain rice in a healthy human nutrition. Agric 11. https://doi.org/10.3390/agriculture11080720
Coelho CP, Santos LA, Rangel RP et al (2016) Rice varieties exhibit different mechanisms for Nitrogen Use Efficiency (NUE). Aust J Crop Sci 10:342–352. https://doi.org/10.21475/ajcs.2016.10.03.p7085
D’Andréa AF, Naves Silva ML, Curi N, Guimarães Guilherme LR (2004) Carbon and nitrogen storage, and inorganic nitrogen forms in a soil under different management systems. Pesqui Agropecu Bras 39:179–186. https://doi.org/10.1590/s0100-204x2004000200012
Demmig-adams B (2006) Tansley review. New Phytol 11–21. https://doi.org/10.1111/j.1469-8137.2006.01835.x
Dinkeloo K, Boyd S, Pilot G (2018) Update on amino acid transporter functions and on possible amino acid sensing mechanisms in plants. Semin Cell Dev Biol 74:105–113. https://doi.org/10.1016/j.semcdb.2017.07.010
Doll H, Andersen B (1981) Preparation of barley storage protein, hordein, for analytical sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Anal Biochem 115:61–66. https://doi.org/10.1016/0003-2697(81)90523-6
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 14. https://doi.org/10.1186/s12284-020-00446-9
Fernandes MS (1984) N-carriers, light and temp influences on free amino acid pool composition of rice plants. Turrialba 34:297–301
Fernandes MS (2014) Integracão de conhecimentos para o uso eficiente de nitrogênio na agricultura. In: Moreira FMS, Kasuya (eds) Fertilidade e Biologia do Solo, vol MCM. SBC, Viçosa-MG, pp 173–196
Frommer WB, Hummel S, Riesmeier JW (1993) Expression cloning in yeast of a cDNA encoding a broad specificity amino acid permease from Arabidopsis thaliana. Proc Natl Acad Sci U S A 90:5944–5948. https://doi.org/10.1073/pnas.90.13.5944
Galili G, Amir R, Fernie AR (2016) The regulation of essential amino acid synthesis and accumulation in plants. Annu Rev Plant Biol 67:153–178. https://doi.org/10.1146/annurev-arplant-043015-112213
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
Guo N, Hu J, Yan M et al (2020) Oryza sativa Lysine-Histidine-type Transporter 1 functions in root uptake and root-to-shoot allocation of amino acids in rice. Plant J 103:395–411. https://doi.org/10.1111/tpj.14742
Guo N, Zhang S, Gu M, Xu G (2021) Function, transport, and regulation of amino acids: What is missing in rice? Crop J 9:530–542. https://doi.org/10.1016/j.cj.2021.04.002
Itoh JI, Nonomura KI, Ikeda K et al (2005) Rice plant development: From zygote to spikelet. Plant Cell Physiol 46:23–47. https://doi.org/10.1093/pcp/pci501
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
Juliano BO (1985) Polysacharides, proteins and lipids of rice. In: Juliano BO (ed) Rice: chemistry and technology. American association of cereal chemists. Saint Paul, Minnesota
Karimi M, Inzé D, Depicker A (2002) GATEWAY vectors for Agrobacterium-mediated plant.pdf. Trends Plant Sci 7:193–195
Lee S (2021) Recent advances on nitrogen use efficiency in rice. Agronomy 11. https://doi.org/10.3390/agronomy11040753
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2-∆∆CT 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
Lu Y, Song Z, Lü K et al (2012) Molecular characterization, expression and functional analysis of the amino acid transporter gene family (OsAATs) in rice. Acta Physiol Plant 34:1943–1962. https://doi.org/10.1007/s11738-012-0995-x
Luo L, Qin R, Liu T et al (2019) OsASN1 plays a critical role in asparagine-dependent rice development. Int J Mol Sci 20:1–15. https://doi.org/10.3390/ijms20010130
Masclaux-Daubresse C, Daniel-Vedele F, Dechorgnat J et al (2010) Nitrogen uptake, assimilation and remobilization in plants: Challenges for sustainable and productive agriculture. Ann Bot 105:1141–1157. https://doi.org/10.1093/aob/mcq028
McCready RM, Guggolz J, Silviera V, Owens HS (1950) Determination of starch and amylose in vegetables. Anal Chem 22:1156–1158. https://doi.org/10.1021/ac60045a016
Muhammad S, Kumazawa K (1974) The absorption, distribution, and redistribution of 15n-labelled ammonium and nitrate nitrogen administered at different growth stages of rice. Soil Sci Plant Nutr 20:47–55. https://doi.org/10.1080/00380768.1974.10433227
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
Perchlik M, Tegeder M (2017) Improving plant nitrogen use efficiency through alteration of amino acid transport processes. Plant Physiol 175:235–247. https://doi.org/10.1104/pp.17.00608
Perchlik M, Tegeder M (2018) Leaf amino acid supply affects photosynthetic and plant nitrogen use efficiency under nitrogen stress. Plant Physiol 178:174–188. https://doi.org/10.1104/pp.18.00597
Pereira EG, Ferreira LM, Fernandes E da et al (2021) Root morphology and ammonium uptake kinetics in two traditional rice varieties submitted to different doses of ammonium nutrition. J Plant Nutr 1–14. https://doi.org/10.1080/01904167.2021.1927088
Qiu X, Xie W, Lian X, Zhang Q (2009) Molecular analyses of the rice glutamate dehydrogenase gene family and their response to nitrogen and phosphorous deprivation. Plant Cell Rep 28:1115–1126. https://doi.org/10.1007/s00299-009-0709-z
R Core Team (2019) R: a language and environment for statistical computing. RFoundation for Statistical Computing, Vienna (Austria). https://www.R-project.org/
Sanders A, Collier R, Trethewy A et al (2009) AAP1 regulates import of amino acids into developing Arabidopsis embryos. Plant J 59:540–552. https://doi.org/10.1111/j.1365-313X.2009.03890.x
Sperandio MVL, Santos LA, Bucher CA et al (2011) Isoforms of plasma membrane H +-ATPase in rice root and shoot are differentially induced by starvation and resupply of NO 3- or NH 4+. Plant Sci 180:251–258. https://doi.org/10.1016/j.plantsci.2010.08.018
Souza SR, Fernandes MS (2018) Nitrogênio. In: Souza SR, Fernandes MS, Santos LA (eds) Nutrição Mineral de Plantas. SBCS, Viçosa-MG, pp 309–375
Tabuchi M, Abiko T, Yamaya T (2007) Assimilation of ammonium ions and reutilization of nitrogen in rice (Oryza sativa L.). J Exp Bot 58:2319–2327. https://doi.org/10.1093/jxb/erm016
Taylor MR, Reinders A, Ward JM (2015) Transport function of rice amino acid permeases (AAPs). Plant Cell Physiol 56:1355–1363. https://doi.org/10.1093/pcp/pcv053
Tedesco MJ, Gianello C, Bissani CA et al (1995) Análises de solo, plantas e outros materiais, vol 5. Ufr, Porto Alegre, p 174
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
Turley RH, Ching TM (1986) Storage protein accumulation in ‘Scio’Barley seed as affected by late foliar applications of nitrogen 1. Crop Sci 26(4):778–782. https://doi.org/10.2135/cropsci1986.0011183X002600040032x
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 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
Xie K, Minkenberg B, Yang Y (2014) Vol 4, Iss 17, Sep 05, 2014. bio-protocol 4:1–9. http://www.bio-protocol.org/e1225
Xu G, Fan X, Miller AJ (2012) Plant nitrogen assimilation and use efficiency. Annu Rev Plant Biol 63:153–182. https://doi.org/10.1146/annurev-arplant-042811-105532
Yemm EW, Cocking EC, Ricketts RE (1955) The determination of amino-acids with ninhydrin. Analyst 80(948):209–213. https://doi.org/10.1039/AN9558000209
Zeng DD, Qin R, Li M et al (2017) The ferredoxin-dependent glutamate synthase (OsFd-GOGAT) participates in leaf senescence and the nitrogen remobilization in rice. Mol Genet Genomics 292:385–395. https://doi.org/10.1007/s00438-016-1275-z
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 7. 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: Conceptualization, Experimental design, Investigation, Formal analysis, Data analysis, Writing – original draft.
JL, CSC: Histochemical analyses, supervision, methodology.
CPCB, CAB, LAS, MSF: Conceptualization, Supervision, Writing – Reviewing and Editing.
Corresponding author
Ethics declarations
Competing interest
The authors declare none.
Additional information
Responsible Editor: Stefano Cesco.
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.
ESM 1
(DOCX 33.0 KB)
Rights and permissions
About this article
Cite this article
Pereira, E.G., Bucher, C.P.C., Bucher, C.A. et al. The amino acid transporter OsAAP1 regulates the fertility of spikelets and the efficient use of N in rice. Plant Soil 480, 507–521 (2022). https://doi.org/10.1007/s11104-022-05598-9
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11104-022-05598-9