Abstract
The grain protein content is an important quality trait in cereals, and the expression level of the OsAAP6 can significantly affect the grain protein content in rice. Through site-directed mutagenesis, we found that the position from −7 to −12 bp upstream of the transcription start site of the OsAAP6 was the functional variation site. By using the yeast single hybrid test, point-to-point in yeast, and the local surface plasmon resonance test, the OsNAC74 was screened and verified to be a regulator upstream of OsAAP6. The OsNAC74 is a constitutively expressed gene whose product is located on the cell membrane. The OsAAP6 and the genes related to the seed storage in the Osnac74 mutants were downregulated, and grain protein content was significantly reduced. In addition, OsNAC74 had a significant impact on quality traits such as grain chalkiness and gel consistency in rice. Although the Osnac74 mutant seeds were relatively small, the individual plant yield was not decreased. Therefore, OsNAC74 is an important regulatory factor with multiple biological functions. This study provides important information for the later use of OsNAC74 gene for molecular design and breeding in rice.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data availability
Not applicable.
References
Alam M, Tan X, Zhang H, Lou G, Yang H, Zhou Y, Hussain A, Bhantana P, Jiang G, He Y (2023) QTL Mining and validation of grain nutritional quality characters in rice (Oryza sativa L.) using two introgression line populations. Agriculture 13(9):1725. https://doi.org/10.3390/agriculture13091725
Chen D, Chai S, McIntyre C, Xue G (2018a) Overexpression of a predominantly root-expressed NAC transcription factor in wheat roots enhances root length, biomass and drought tolerance. Plant Cell Rep 37(2):225–237. https://doi.org/10.1007/s00299-017-2224-y
Chen P, Lou G, Wang Y, Chen J, Chen W, Fan Z, Liu Q, Sun B, Mao X, Yu H, Jiang L, Zhang J, Lv S, Xing J, Pan D, Li C, He Y (2022) The genetic basis of grain protein content in rice by genome-wide association analysis. Mol Breed 43:1. https://doi.org/10.1007/s11032-022-01347-z
Chen P, Shen Z, Ming L, Li Y, Dan W, Lou G, Peng B, Wu B, Li Y, Zhao D, Gao G, Zhang Q, Xiao J, Li X, Wang G, He Y (2018b) Genetic basis of variation in rice seed storage protein (albumin, globulin, prolamin, and glutelin) content revealed by genome-wide association analysis. Front Plant Sci 9:612. https://doi.org/10.3389/fpls.2018.00612
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(1):2. https://doi.org/10.1186/s12284-020-00446-9
Fischer W, Kwart M, Hummel S, Frommer W (1995) Aubstrate specificity and expression profile of amino acid transporters (aaps) in Arabidopsis. J Biol Chem 270(27):16315–16320. https://doi.org/10.1074/jbc.270.27.16315
Frommer W, Hummel S, Riesmeier J (1993) Expression cloning in yeast of a cdna encoding a broad specificity amino acid permease from Arabidopsis thaliana. Proc Natl Acad Sci USA 90(13):5944–5948. https://doi.org/10.1073/pnas.90.13.5944
Fujita M, Fujita Y, Maruyama K, Seki M, Hiratsu K, Ohme-Takagi M, Tran L, Yamaguchi-Shinozaki K, Shinozaki K (2004) A dehydration-induced NAC protein RD26 is involved in a novel ABA-dependent stress-signaling pathway. Plant J 39(6):863–876. https://doi.org/10.1111/j.1365-313x.2004.02171.x
Han T, Yan J, Xiang Y, Zhang A (2021) Phosphorylation of ZmNAC84 at Ser-113 enhances the drought tolerance by directly modulating ZmSOD2 expression in maize. Biochem Biophys Res Commun 567(27):86–91. https://doi.org/10.1016/j.bbrc.2021.06.026
Hao Y, Wei W, Song Q, Chen H, Zhang Y, Wang F, Zou H, Lei G, Tian A, Zhang W, Ma B, Zhang J, Chen S (2011) Soybean NAC transcription factors promote abiotic stress tolerance and lateral root formation in transgenic plants. Plant J 68(2):302–313. https://doi.org/10.1111/j.1365-313x.2011.04687.x
He Y, Wang S, Ding Y (2013) Identification of novel glutelin subunits and a comparison of glutelin composition between japonica and indica rice (Oryza sativa L.). J Cereal Sci 57:362–371. https://doi.org/10.1016/j.jcs.2012.12.009
Hong Y, Zhang H, Huang L, Li D, Song F (2016) Overexpression of a stress-responsive NAC transcription factor gene ONAC022 improves drought and salt tolerance in rice. Front Plant Sci 7:4. https://doi.org/10.3389/fpls.2016.00004
Hu H, Dai M, Yao J, Xiao B, Li X, Zhang Q, Xiong L (2006) Overexpressing a NAM ATAF and CUC (NAC) transcription factor enhances drought resistance and salt tolerance in rice. Proc Natl Acad Sci USA 103(35):12987–12992. https://doi.org/10.1073/pnas.0604882103
Hunt E, Gattolin S, Newbury H, Bale J, Tseng H, Barrett D, Pritchard J (2010) A mutation in amino acid permease AAP6 reduces the amino acid content of the Arabidopsis sieve elements but leaves aphid herbivores unaffected. J Exp Bot 61(1):55–64. https://doi.org/10.1093/jxb/erp274
Ji Y, Huang W, Wu B, Fang Z, Wang X (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(16):4763–4777. https://doi.org/10.1093/jxb/eraa256
Kato H, Motomura T, Komeda Y, Saito T, Kato A (2010) Overexpression of the NAC transcription factor family gene ANAC036 results in a dwarf phenotype in Arabidopsis thaliana. J Plant Physiol 167(7):571–577. https://doi.org/10.1016/j.jplph.2009.11.004
Kim Y, Kim S, Park J, Park H, Lim M, Chua N, Park C (2006) A membrane-bound NAC transcription factor regulates cell division in Arabidopsis. Plant Cell 18(11):3132–3144. https://doi.org/10.1105/tpc.106.043018
Lee Y, Foster J, Chen J, Voll L, Weber A, Tegeder M (2007) AAP1 transports uncharged amino acids into roots of Arabidopsis. Plant J 50(2):305–319. https://doi.org/10.1111/j.1365-313x.2007.03045.x
Liu Z, Jiang S, Jiang L, Li W, Tang Y, He W, Wang M, Xing J, Cui Y, Lin Q, Yu F, Wang L (2022) Transcription factor OsSGL is a regulator of starch synthesis and grain quality in rice. J Exp Bot 73:3417–3430. https://doi.org/10.1093/jxb/erac068
Lu K, Wu B, Wang J, Zhu W, Nie H, Qian J, Huang W, Fang Z (2018) Blocking amino acid transporter OsAAP3 improves grain yield by promoting outgrowth buds and increasing tiller number in rice. Plant Biotechnol J 16(10):1710–1722. https://doi.org/10.1111/pbi.12907
Luke L, Thirruvithamkottil M, Thomas M, Kuruvilla L (2017) Expression of NAC transcription factor is altered under intermittent drought stress and re-watered conditions in hevea brasiliensis. J Plant Biotech 44:142–148. https://doi.org/10.5010/JPB.2017.44.2.142
Nakashima K, Tran L, Van Nguyen D, Fujita M, Maruyama K, Todaka D, Ito Y, Hayashi N, Shinozaki K, Yamaguchi-Shinozaki K (2007) Functional analysis of a NAC-type transcription factor OsNAC6 involved in abiotic and biotic stress-responsive gene expression in rice. Plant J 51(4):617–630. https://doi.org/10.1111/j.1365-313x.2007.03168.x
Okumoto S, Koch W, Tegeder M, Fischer W, Biehl A, Leister D, Stierhof Y, Frommer W (2004) Root phloem-specific expression of the plasma membrane amino acid proton co-transporter AAP3. J Exp Bot 55(406):2155–2168. https://doi.org/10.1093/jxb/erh233
Okumoto S, Schmidt R, Tegeder M, Fischer W, Rentsch D, Frommer W, Koch W (2002) High affinity amino acid transporters specifically expressed in xylem parenchyma and developing seeds of Arabidopsis. J Biol Chem 277(47):45338–45346. https://doi.org/10.1074/jbc.m207730200
Peng B, Kong D, Huang Y, Jia Y, Ma C, Song H, Sun Y, He L, Pang R, Song X, Li H, Liu L, Huang Y, Zhou Q, Li J, Song S (2018) Bioinformatics analysis of the structure and function of OsAAP6 gene in rice. J Southwest Agri 31(3):429–436. https://doi.org/10.16213/j.cnki.scjas.2018.3.001
Peng B, Kong H, Li Y, Wang L, Zhong M, Sun L, Gao G, Zhang Q, Luo L, Wang G, Xie W, Chen J, Yao W, Peng Y, Lei L, Lian X, Xiao J, Xu C, Li X, He Y (2014b) OsAAP6 functions as an important regulator of grain protein content and nutritional quality in rice. Nat Commun 5:4847. https://doi.org/10.1038/ncomms5847
Peng B, Wang L, Fan C, Jiang G, Luo L, Li Y, He Y (2014a) Comparative mapping of chalkiness components in rice using five populations across two environments. BMC Genet 15(1):49. https://doi.org/10.1186/1471-2156-15-49
Pereira E, Bucher C, Bucher C, Santos L, Lerin J, Catarina C, Fernandes M (2022) The amino acid transporter OsAAP1 regulates the fertility of spikelets and the efficient use of N in rice. Plant Soil 480:507–521. https://doi.org/10.1007/s11104-022-05598-9
Ray S, Basnet A, Bhattacharya S, Banerjee A, Biswas K (2023) A Comprehensive analysis of NAC gene family in Oryza Sativa Japonica: a structural and functional genomics approach. J Biomol Struct Dyn 41(3):856–870. https://doi.org/10.1080/07391102.2021.2014968
Ren D, Ding C, Qian Q (2023) Molecular bases of rice grain size and quality for optimized productivity. Sci Bull 68(3):314–350. https://doi.org/10.1016/j.scib.2023.01.026
Ren Y, Huang Z, Jiang H, Wang Z, Wu F, Xiong Y, Yao J (2021) A heat stress responsive NAC transcription factor heterodimer plays key roles in rice grain filling. J Exp Bot 72(8):2947–2964. https://doi.org/10.1093/jxb/erab027
Riechmann J, Heard J, Martin G, Reuber L, Jiang C, Keddie J, Adam L, Pineda O, Ratcliffe O, Samaha R, Creelman R, Pilgrim M, Broun P, Zhang J, Ghandehari D, Sherman B, Yu G (2000) Arabidopsis transcription factors: genome-wide comparative analysis among eukaryotes. Science 290(5499):2105–2110. https://doi.org/10.1126/science.290.5499.2105
Rogers J, Bulyk M (2018) Diversification of transcription factor-DNA interactions and the evolution of gene regulatory networks. Wiley Interdiscip Rev Syst Biol Med 10(5):e1423. https://doi.org/10.1002/wsbm.1423
Schmidt R, Stransky H, Koch W (2007) The amino acid permease AAP8 is important for early seed development in Arabidopsis thaliana. Planta 226(4):805–813. https://doi.org/10.1007/s00425-007-0527-x
Sperotto R, Ricachenevsky F, Duarte G, Boff T, Lopes K, Sperb E, Grusak M, Fett J (2009) Identification of up-regulated genes in flag leaves during rice grain filling and characterization of OsNAC5 a new ABA-dependent transcription factor. Planta 230(5):985–1002. https://doi.org/10.1007/s00425-009-1000-9
Svennerstam H, Ganeteg U, Näsholm T (2008) Root uptake of cationic amino acids by arabidopsis depends on functional expression of amino acid permease 5. New Phytol 180(3):620–630. https://doi.org/10.1111/j.1469-8137.2008.02589.x
Taylor M, Reinders A, Ward J (2015) Transport function of rice amino acid permeases (AAPs). Plant Cell Physiol 56(7):1355–1363. https://doi.org/10.1093/pcp/pcv053
Tegeder M (2014) Transporters involved in source to sink partitioning of amino acids and ureides: opportunities for crop improvement. J Exp Bot 65(7):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(1):35–53. https://doi.org/10.1111/nph.14876
Tran L, Nakashima K, Sakuma Y, Simpson S, Fujita Y, Maruyama K, Fujita M, Seki M, Shinozaki K, Yamaguchi-Shinozaki K (2004) Isolation and functional analysis of Arabidopsis stress-inducible NAC transcription factors that bind to a drought-responsive cis-element in the early responsive to dehydration stress 1 promoter. Plant Cell 16(9):2481–2498. https://doi.org/10.1105/tpc.104.022699
Wang G, Yuan Z, Zhang P, Liu Z, Wang T, Wei L (2020c) Genome-wide analysis of NAC transcription factor family in maize under drought stress and rewatering. Physiol Mol Biol Plants 26(4):705–717. https://doi.org/10.1007/s12298-020-00770-w
Wang J, Chen Z, Zhang Q, Meng S, Wei C (2020a) The NAC transcription factors OsNAC20 and OsNAC26 regulate starch and storage protein synthesis. Plant Physiol 184(4):1775–1791. https://doi.org/10.1104/pp.20.00984
Wang J, Wu B, Lu K, Wei Q, Qian J, Chen Y, Fang Z (2019) The amino acid permease 5 (OsAAP5) regulates tiller number and grain yield in rice. Plant Physiol 180(2):1031–1045. https://doi.org/10.1104/pp.19.00034
Wang S, Yang Y, Guo M, Zhong C, Yan C, Sun S (2020b) Targeted mutagenesis of amino acid transporter genes for rice quality improvement using the CRISPR/Cas9 system. Crop J 3:457–464. https://doi.org/10.1016/j.cj.2020.02.005
Xia D, Wang Y, Shi Q, Wu B, Yu X, Zhang C, Li Y, Fu P, Li M, Zhang Q, Liu Q, Gao G, Zhou H, He Y (2022) Effects of Wx genotype, nitrogen fertilization, and temperature on rice grain quality. Front Plant Sci 13(3):901541. https://doi.org/10.3389/fpls.2022.901541
Yang G, Wei X, Fang Z (2022b) Melatonin mediates axillary bud outgrowth by improving nitrogen assimilation and transport in rice. Front Plant Sci 13:900262. https://doi.org/10.3389/fpls.2022.900262
Yang X, Kim M, Ha J, Lee S (2019) Overexpression of the soybean NAC gene GmNAC109 increases lateral root formation and abiotic stress tolerance in transgenic Arabidopsis plants. Front Plant Sci 10:1036. https://doi.org/10.3389/fpls.2019.01036
Yang X, Yang G, Wei X, Huang W, Fang Z (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
Yang Y, Shen Z, Li Y, Xu C, Xia H, Zhuang H, Sun S, Guo M, Yan C (2022a) Rapid improvement of rice eating and cooking quality through gene editing toward glutelin as target. J Integr Plant Biol 64(10):1860–1865. https://doi.org/10.1111/jipb.13334
Yu X, Liu Y, Wang S, Tao Y, Wang Z, Shu Y, Peng H, Mijiti A, Wang Z, Zhang H, Ma H (2016) CarNAC4 a NAC-type chickpea transcription factor conferring enhanced drought and salt stress tolerances in Arabidopsis. Plant Cell Rep 35(3):613–627. https://doi.org/10.1007/s00299-015-1907-5
Zhang L, Tan Q, Lee R, Trethewy A, Lee Y, Tegede M (2010) Altered xylem-phloem transfer of amino acids affects metabolism and leads to increased seed yield and oil content in Arabidopsis. Plant Cell 22(11):3603–3620. https://doi.org/10.1105/tpc.110.073833
Zhang X, Long Y, Chen X, Zhang B, Xin Y, Li L, Cao S, Liu F, Wang Z, Huang H, Zhou D, Xia J (2021) A NAC transcription factor OsNAC3 positively regulates ABA response and salt tolerance in rice. BMC Plant Biol 21(1):546. https://doi.org/10.1186/s12870-021-03333-7
Zhang Y, Su J, Duan S, Ao Y, Dai J, Liu J, Wang P, Li Y, Liu B, Feng D, Wang J, Wang H (2011) A highly efficient rice green tissue protoplast system for transient gene expression and studying light/chloroplast-related processes. Plant Methods 7(1):30. https://doi.org/10.1186/1746-4811-7-30
Zhao M, Lin Y, Chen H (2020) Improving nutritional quality of rice for human health. Theor Appl Genet 133(5):1397–1413. https://doi.org/10.1007/s00122-019-03530-x
Zhong M, Wang L, Yuan D, Luo L, Xu C, He Y (2011) Identification of QTL affecting protein and amino acid contents in rice. Rice Sci 18(3):187–195. https://doi.org/10.1016/s1672-6308(11)60026-7
Zhong R, Lee C, Ye Z (2009) Functional characterization of poplar wood-associated NAC domain transcription factors. Plant Physiol 152(2):1044–1055. https://doi.org/10.1104/pp.109.148270
Funding
This work was financially supported by the National Natural Science Foundation of China (U2004141, 31801332), Key Project of Science and Technology in Henan Province (222102110141; 222102110115; 212102110249), Xinyang Innovation Project (20210007), Graduate Research Innovation Fund Project of XYNU (2022KYJJ066, 2022KYJJ068), and Scientific Research Fund Project of XYNU (2022-DXS-131).
Author information
Authors and Affiliations
Contributions
Bo Peng and Quanxiu Wang designed the experiments. Bo Peng, Xiaoyu Sun, and Xiayu Tian carried out most of the experiments and analyzed the data. Dongyan Kong and Lulu He contributed to the experiments and formal analysis. Juan Peng and Yan Liu contributed to the experiments. Gui-Ying Guo, Yanfang Sun, and Ruihua Pang contributed to the experiments. Wei Zhou and Jinhui Zhao contributed to the experiments and analyzed the data. Bo Peng and Quanxiu Wang wrote the manuscript.
Corresponding authors
Ethics declarations
Ethics approval and consent to participate
Not applicable.
Consent for publication
Not applicable.
Conflict of interest
The authors declare no competing interests.
Additional information
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary information
ESM 1
(DOCX 2060 kb)
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
Peng, B., Sun, X., Tian, X. et al. OsNAC74 affects grain protein content and various biological traits by regulating OsAAP6 expression in rice. Mol Breeding 43, 87 (2023). https://doi.org/10.1007/s11032-023-01433-w
Received:
Accepted:
Published:
DOI: https://doi.org/10.1007/s11032-023-01433-w