Abstract
Main conclusion
OsTST1 affects yield and development and mediates sugar transportation of plants from source to sink in rice, which influences the accumulation of intermediate metabolites from tricarboxylic acid cycle indirectly.
Abstract
Tonoplast sugar transporters (TSTs) are essential for vacuolar sugar accumulation in plants. Carbohydrate transport across tonoplasts maintains the metabolic balance in plant cells, and carbohydrate distribution is crucial to plant growth and productivity. Large plant vacuoles store high concentrations of sugars to meet plant requirements for energy and other biological processes. The abundance of sugar transporter affects crop biomass and reproductive growth. However, it remains unclear whether the rice (Oryza sativa L.) sugar transport protein OsTST1 affects yield and development. In this study, we found that OsTST1 knockout mutants generated via CRISPR/Cas9 exhibited slower development, smaller seeds, and lower yield than wild type (WT) rice plants. Notably, plants overexpressing OsTST1 showed the opposite effects. Changes in rice leaves at 14 days after germination (DAG) and at 10 days after flowering (DAF) suggested that OsTST1 affected the accumulation of intermediate metabolites from the glycolytic pathway and the tricarboxylic acid (TCA) cycle. The modification of the sugar transport between cytosol and vacuole mediated by OsTST1 induces deregulation of several genes including transcription factors (TFs). In summary, no matter the location of sucrose and sink is, these preliminary results revealed that OsTST1 was important for sugar transport from source to sink tissues, thus affecting plant growth and development.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data availability
All data related to this manuscript can be found within this paper and its Supplementary data.
Abbreviations
- DAG:
-
Days after germination
- DAF:
-
Days after flowering
- DEG:
-
Differentially expressed gene
- FC:
-
Fold-change
- OE:
-
Overexpression
- TST:
-
Tonoplast sugar transporters
References
Abelenda JA, Bergonzi S, Oortwijn M, Sonnewald S, Du M, Visser RGF, Sonnewald U, Bachem CWB (2019) Source-sink regulation is mediated by interaction of an FT homolog with a SWEET protein in potato. Curr Biol 29(7):1178–1186. https://doi.org/10.1016/j.cub.2019.02.018
Alexa A, Rahnenfuhrer J, Lengauer T (2006) Improved scoring of functional groups from gene expression data by decorrelating GO graph structure. Bioinformatics 22(13):1600–1607. https://doi.org/10.1093/bioinformatics/btl140
Alseekh S, Aharoni A, Brotman Y, Contrepois K, D’Auria J, Ewald J, Ewald JC, Fraser PD, Giavalisco P, Hall RD, Heinemann M, Link H, Luo J, Neumann S, Nielsen J, Perez de Souza L, Saito K, Sauer U, Schroeder FC, Schuster S, Siuzdak G, Skirycz A, Sumner LW, Snyder MP, Tang H, Tohge T, Wang Y, Wen W, Wu S, Xu G, Zamboni N, Fernie AR (2021) Mass spectrometry-based metabolomics: a guide for annotation, quantification and best reporting practices. Nat Methods 18(7):747–756. https://doi.org/10.1038/s41592-021-01197-1
Anders S, Pyl PT, Huber W (2015) HTSeq–a Python framework to work with high-throughput sequencing data. Bioinformatics 31(2):166–169. https://doi.org/10.1093/bioinformatics/btu638
Bi YM, Zhang Y, Signorelli T, Zhao R, Zhu T, Rothstein S (2005) Genetic analysis of Arabidopsis GATA transcription factor gene family reveals a nitrate-inducible member important for chlorophyll synthesis and glucose sensitivity. Plant J 44(4):680–692. https://doi.org/10.1111/j.1365-313X.2005.02568.x
Chao Q, Gao Z-F, Zhang D, Zhao B-G, Dong F-Q, Fu C-X, Liu L-J, Wang B-C (2019) The developmental dynamics of the Populus stem transcriptome. Plant Biotechnol J 17(1):206–219. https://doi.org/10.1111/pbi.12958
Chen L, Song Y, Li S, Zhang L, Zou C, Yu D (2012) The role of WRKY transcription factors in plant abiotic stresses. Biochim Biophys Acta 1819(2):120–128. https://doi.org/10.1016/j.bbagrm.2011.09.002
Chen Q, Xu X, Xu D, Zhang H, Zhang C, Li G (2019) WRKY18 and WRKY53 coordinate with histone acetyltransferase1 to regulate rapid responses to sugar. Plant Physiol 180(4):2212–2226. https://doi.org/10.1104/pp.19.00511
Chen Q, Zhang J, Li G (2022a) Dynamic epigenetic modifications in plant sugar signal transduction. Trends Plant Sci 27(4):379–390. https://doi.org/10.1016/j.tplants.2021.10.009
Chen Y, Luo L, Xu F, Xu X, Bao J (2022b) Carbohydrate repartitioning in the rice starch branching enzyme IIb mutant stimulates higher resistant starch content and lower seed weight revealed by multiomics analysis. J Agric Food Chem 70(31):9802–9816. https://doi.org/10.1021/acs.jafc.2c03737
Cheng J, Wen S, **ao S, Lu B, Ma M, Bie Z (2018a) Overexpression of the tonoplast sugar transporter CmTST2 in melon fruit increases sugar accumulation. J Exp Bot 69(3):511–523. https://doi.org/10.1093/jxb/erx440
Cheng R, Cheng Y, Lu J, Chen J, Wang Y, Zhang S, Zhang H (2018b) The gene PbTMT4 from pear (Pyrus bretschneideri) mediates vacuolar sugar transport and strongly affects sugar accumulation in fruit. Physiol Plant 164(3):307–319. https://doi.org/10.1111/ppl.12742
Cho JI, Burla B, Lee DW, Ryoo N, Hong SK, Kim HB, Eom JS, Choi SB, Cho MH, Bhoo SH, Hahn TR, Neuhaus HE, Martinoia E, Jeon JS (2010) Expression analysis and functional characterization of the monosaccharide transporters, OsTMTs, involving vacuolar sugar transport in rice (Oryza sativa). New Phytol 186(3):657–668. https://doi.org/10.1111/j.1469-8137.2010.03194.x
Dhungana SR, Braun DM (2022) Genomic analyses of SUT and TST sugar transporter families in low and high sugar accumulating sugarcane species (Saccharum spontaneum and Saccharum officinarum). Tropical Plant Biol 15(3):181–196. https://doi.org/10.1007/s12042-022-09315-9
Feng Y, Yuan X, Wang Y, Yang Y, Zhang M, Yu H, Xu Q, Wang S, Niu X, Wei X (2021) Validation of a QTL for grain size and weight using an introgression line from a cross between Oryza sativa and Oryza minuta. Rice (n y) 14(1):43. https://doi.org/10.1186/s12284-021-00472-1
Ge L, Chen H, Jiang JF, Zhao Y, Xu ML, Xu YY, Tan KH, Xu ZH, Chong K (2004) Overexpression of OsRAA1 causes pleiotropic phenotypes in transgenic rice plants, including altered leaf, flower, and root development and root response to gravity. Plant Physiol 135(3):1502–1513. https://doi.org/10.1104/pp.104.041996
Hedrich R, Sauer N, Neuhaus HE (2015) Sugar transport across the plant vacuolar membrane: nature and regulation of carrier proteins. Curr Opin Plant Biol 25:63–70. https://doi.org/10.1016/j.pbi.2015.04.008
Huang T, Yu D, Wang X (2021a) VvWRKY22 transcription factor interacts with VvSnRK1.1/VvSnRK1.2 and regulates sugar accumulation in grape. Biochem Biophys Res Commun 554:193–198. https://doi.org/10.1016/j.bbrc.2021.03.092
Huang X, Zhang Y, Wang L, Dong X, Hu W, Jiang M, Chen G, An G, **ong F, Wu YJFiPS, (2021b) OsDOF11 affects nitrogen metabolism by sucrose transport signaling in rice (Oryza sativa L). Front Plant Sci 12:703034. https://doi.org/10.3389/fpls.2021.703034
Jeong JS, Kim YS, Baek KH, Jung H, Ha SH, Do Choi Y, Kim M, Reuzeau C, Kim JK (2010) Root-specific expression of OsNAC10 improves drought tolerance and grain yield in rice under field drought conditions. Plant Physiol 153(1):185–197. https://doi.org/10.1104/pp.110.154773
Kirimura K, Kobayashi K, Yoshioka I (2019) Decrease of citric acid produced by Aspergillus niger through disruption of the gene encoding a putative mitochondrial citrate-oxoglutarate shuttle protein. Biosci Biotechnol Biochem 83(8):1538–1546. https://doi.org/10.1080/09168451.2019.1574205
Lalonde S, Wipf D, Frommer WB (2004) Transport mechanisms for organic forms of carbon and nitrogen between source and sink. Annu Rev Plant Biol 55:341–372. https://doi.org/10.1146/annurev.arplant.55.031903.141758
Lemoine R, Camera SL, Atanassova R, Dédaldéchamp F, Allario T, Pourtau N, Bonnemain J-L, Laloi M, Coutos-Thévenot P, Maurousset L, Faucher M, Girousse C, Lemonnier P, Parrilla J, Durand M (2013) Source-to-sink transport of sugar and regulation by environmental factors. Front Plant Sci 4:272. https://doi.org/10.3389/fpls.2013.00272
Li P, Wang L, Liu H, Yuan M (2022) Impaired SWEET-mediated sugar transportation impacts starch metabolism in develo** rice seeds. Crop J 10(1):98–108. https://doi.org/10.1016/j.cj.2021.04.012
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods 25(4):402–408. https://doi.org/10.1006/meth.2001.1262
Love MI, Huber W, Anders S (2014) Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2. Genome Biol 15(12):550. https://doi.org/10.1186/s13059-014-0550-8
Lu B, Yuan Y, Zhang C, Ou J, Zhou W, Lin QJPS (2005) Modulation of key enzymes involved in ammonium assimilation and carbon metabolism by low temperature in rice (Oryza sativa L.) roots. Plant Sci 169(2):295–302
Meng X, Liu H, Peng L, He W, Li S (2022) Potential clinical applications of alpha-ketoglutaric acid in diseases (review). Mol Med Rep 25(5):151. https://doi.org/10.3892/mmr.2022.12667
Nino-Gonzalez M, Novo-Uzal E, Richardson DN, Barros PM, Duque P (2019) More transporters, more substrates: The Arabidopsis major facilitator superfamily revisited. Mol Plant 12(9):1182–1202. https://doi.org/10.1016/j.molp.2019.07.003
Popova TN, Pinheiro de Carvalho MA (1998) Citrate and isocitrate in plant metabolism. Biochim Biophys Acta 1364(3):307–325. https://doi.org/10.1016/s0005-2728(98)00008-5
Price J, Laxmi A, St Martin SK, Jang JC (2004) Global transcription profiling reveals multiple sugar signal transduction mechanisms in Arabidopsis. Plant Cell 16(8):2128–2150. https://doi.org/10.1105/tpc.104.022616
Scofield GN, Hirose T, Gaudron JA, Furbank RT, Upadhyaya NM, Ohsugi R (2002) Antisense suppression of the rice sucrose transporter gene, OsSUT1, leads to impaired grain filling and germination but does not affect photosynthesis. Funct Plant Biol 29(7):815–826. https://doi.org/10.1071/PP01204
Shi JH, Lu JY, Chen HY, Wei CC, Xu X, Li H, Bai Q, **a FZ, Lam SM, Zhang H, Shi YN, Cao D, Chen L, Shui G, Yang X, Lu Y, Chen YX, Zhang WJ (2020) Liver ChREBP protects against fructose-induced glycogenic hepatotoxicity by regulating L-type pyruvate kinase. Diabetes 69(4):591–602. https://doi.org/10.2337/db19-0388
Singh A, Roychoudhury A, Samanta S, Banerjee A (2020) Fluoride stress-mediated regulation of tricarboxylic acid cycle and sugar metabolism in rice seedlings in absence and presence of exogenous calcium. J Plant Growth Regul 40(4):1579–1593. https://doi.org/10.1007/s00344-020-10202-4
Smith KM, Alkhateeb FL, Brennan K, Rainville PD, Walter. TH (2021) Separation of pentose phosphate pathway, glycolysis, and energy metabolites using an ACQUITY premier system with an atlantis premier BEH Z-HILIC column. Waters Application Note 720007411
Sun C, Palmqvist S, Olsson H, Boren M, Ahlandsberg S, Jansson C (2003) A novel WRKY transcription factor, SUSIBA2, participates in sugar signaling in barley by binding to the sugar-responsive elements of the iso1 promoter. Plant Cell 15(9):2076–2092. https://doi.org/10.1105/tpc.014597
Taiz L, Zeiger E, Møller IM, Murphy A (2015) Plant physiology and development, 6th edn. Sinauer Associates Incorporated, Sunderland, CT
Thalor SK, Berberich T, Lee SS, Yang SH, Zhu X, Imai R, Takahashi Y, Kusano T (2012) Deregulation of sucrose-controlled translation of a bZIP-type transcription factor results in sucrose accumulation in leaves. PLoS ONE 7(3):e33111. https://doi.org/10.1371/journal.pone.0033111
Umer MJ, Bin Safdar L, Gebremeskel H, Zhao S, Yuan P, Zhu H, Kaseb MO, Anees M, Lu X, He N, Gong C, Liu W (2020) Identification of key gene networks controlling organic acid and sugar metabolism during watermelon fruit development by integrating metabolic phenotypes and gene expression profiles. Hortic Res 7(1):193. https://doi.org/10.1038/s41438-020-00416-8
Walter TH, Alden BA, Berthelette K, Field JA, Lawrence NL, McLaughlin J, Patel AV (2022) Characterization of a highly stable zwitterionic hydrophilic interaction chromatography stationary phase based on hybrid organic-inorganic particles. J Sep Sci 45(8):1389–1399. https://doi.org/10.1002/jssc.202100859
Wang J, Liu H, Zhao C, Tang H, Mu Y, Xu Q, Deng M, Jiang Q, Chen G, Qi P, Wang J, Jiang Y, Chen S, Wei Y, Zheng Y, Lan X, Ma J (2022) Map** and validation of major and stable QTL for flag leaf size from tetraploid wheat. Plant Genome 15(4):e20252. https://doi.org/10.1002/tpg2.20252
Wiese A, Elzinga N, Wobbes B, Smeekens S (2004) A conserved upstream open reading frame mediates sucrose-induced repression of translation. Plant Cell 16(7):1717–1729. https://doi.org/10.1105/tpc.019349
Wingenter K, Schulz A, Wormit A, Wic S, Trentmann O, Hoermiller II, Heyer AG, Marten I, Hedrich R, Neuhaus HE (2010) Increased activity of the vacuolar monosaccharide transporter TMT1 alters cellular sugar partitioning, sugar signaling, and seed yield in Arabidopsis. Plant Physiol 154(2):665–677. https://doi.org/10.1104/pp.110.162040
Wormit A, Trentmann O, Feifer I, Lohr C, Tjaden J, Meyer S, Schmidt U, Martinoia E, Neuhaus HE (2006) Molecular identification and physiological characterization of a novel monosaccharide transporter from Arabidopsis involved in vacuolar sugar transport. Plant Cell 18(12):3476–3490. https://doi.org/10.1105/tpc.106.047290
Yang J, Luo D, Yang B, Frommer WB, Eom JS (2018) SWEET11 and 15 as key players in seed filling in rice. New Phytol 218(2):604–615. https://doi.org/10.1111/nph.15004
Yoon J, Cho LH, Tun W, Jeon JS, An G (2021) Sucrose signaling in higher plants. Plant Sci 302:110703. https://doi.org/10.1016/j.plantsci.2020.110703
Yu SM, Lo SF, Ho TD (2015) Source-sink communication: regulated by hormone, nutrient, and stress cross-signaling. Trends Plant Sci 20(12):844–857. https://doi.org/10.1016/j.tplants.2015.10.009
Yu J, **ong H, Zhu X, Zhang H, Li H, Miao J, Wang W, Tang Z, Zhang Z, Yao G, Zhang Q, Pan Y, Wang X, Rashid MAR, Li J, Gao Y, Li Z, Yang W, Fu X, Li Z (2017) OsLG3 contributing to rice grain length and yield was mined by Ho-LAMap. BMC Biol. https://doi.org/10.1186/s12915-017-0365-7
Zhang J, Li J, Wang X, Chen J (2011) OVP1, a vacuolar H+-translocating inorganic pyrophosphatase (V-PPase), overexpression improved rice cold tolerance. Plant Physiol Biochem 49(1):33–38. https://doi.org/10.1016/j.plaphy.2010.09.014
Zhang J-S, Cai Y, Chen X, **e K, **ng Q, Wu Y, Li J, Du C, Sun Z, Guo Z (2014) Dlf1, a WRKY transcription factor, is involved in the control of flowering time and plant height in Rice. PLoS ONE 9(7):e102529. https://doi.org/10.1371/journal.pone.0102529
Zhang S, Zhang Y, Li K, Yan M, Zhang J, Yu M, Tang S, Wang L, Qu H, Luo L, Xuan W, Xu G (2021) Nitrogen mediates flowering time and nitrogen use efficiency via floral regulators in rice. Curr Biol 31(4):671-683e675. https://doi.org/10.1016/j.cub.2020.10.095
Zhao H, Sun S, Zhang L, Yang J, Wang Z, Ma F, Li M (2020) Carbohydrate metabolism and transport in apple roots under nitrogen deficiency. Plant Physiol Biochem 155:455–463. https://doi.org/10.1016/j.plaphy.2020.07.037
Zhao D, Zhang C, Li Q, Liu Q (2022) Genetic control of grain appearance quality in rice. Biotechnol Adv 60:108014. https://doi.org/10.1016/j.biotechadv.2022.108014
Zhu L, Li B, Wu L, Li H, Wang Z, Wei X, Ma B, Zhang Y, Ma F, Ruan Y-L, Li M (2020) MdERDL6 -mediated glucose efflux to the cytosol promotes sugar accumulation in the vacuole through up-regulating TSTs in apple and tomato. Proc Natl Acad Sci USA 118(1):e2022788118. https://doi.org/10.1073/pnas.2022788118
Acknowledgements
This study was supported by grants from the Ministry of Agriculture of China for Transgenic Research (2016ZX08009002-006).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
There are no moral and ethical issues or conflicts to declare in this paper.
Additional information
Communicated by Dorothea Bartels.
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.
425_2023_4160_MOESM4_ESM.docx
Supplementary file4 Evolutionary analysis of tonoplast sugar transporters (TSTs) in 9 species (a), expression levels in different rice tissues at 10 d after flowering (DAF) (b) and ostst1 mutants (c-f). a Phylogenetic tree showing relationships between TSTs in representative monocotyledons, dicotyledons, and algae, namely Arabidopsis thaliana, Brassica napus, Oryza sativa, Panicum virgatum, Populus trichocarpa, Sorghum bicolor, Setaria italica, Triticum aestivum, and Zea mays. b At 10 d after flowering (DAF), OsTST1 was expressed at different levels between tissues (namely the flag leaves, leaves, glumes, stems, panicle axis, and roots). OsTST1 was primarily expressed in the flag leaves. c-e Expression levels of four OsTST genes were measured in transgenic homozygous rice via quantitative reverse transcription (qRT)-PCR. Levels of OsTST1 decreased to less than 40% of wild type (WT) levels in the three mutants. Expression levels of the other three OsTST isoforms were comparable to those in the WT (DOCX 983 KB)
425_2023_4160_MOESM5_ESM.docx
Supplementary file5 OsTST1 overexpression (OE) increased rice grain size and plant height. a-d Comparison of 100-grain weight, seed length, seed thickness, and seed width between wild type (WT), OE#8, OE#14, and OE#27 plants. Seeds were selected at random from each transgenic line. *P ≤ 0.05, **P ≤ 0.01 (Student’s t-test). n = 30 for seed length, thickness, and width. e Representative images of plants at 14 d after germination (DAG). OE plant lines were clearly taller than WT plants. Scale bar = 5 cm. f Quantification of plant height at 14 DAG. OE plants were significantly taller than WT plants, and OE14 plants were the tallest among the OE lines. *P ≤ 0.05, **P ≤ 0.01 (Student’s t-test). n = 10 (DOCX 605 KB)
425_2023_4160_MOESM6_ESM.docx
Supplementary file6 Heat maps showing levels of 43 metabolites in wild type (WT) and ostst1#3 plants. Metabolites were extracted from young leaves at 14 d after germination (DAG). They were listed in descending order of content in the ostst1#3 mutants. There were five replicates for each genotype (DOCX 544 KB)
425_2023_4160_MOESM7_ESM.docx
Supplementary file7 Heat maps showing levels of 32 metabolites in wild type (WT) and ostst1#3 plants. Metabolites were extracted from flag leaves at 10 d after flowering (DAF). They were listed in descending order of content in the ostst1#3 mutants. There were five replicates for each genotype (DOCX 396 KB)
425_2023_4160_MOESM8_ESM.docx
Supplementary file8 Heat maps showing differentially expressed genes (DEGs) between wild type (WT) and ostst1#3 plants at 14 d after germination (DAG). DEGs were identified in young leaves with a false discovery rate (FDR) threshold of < 0.01. There were 100 DEGs between the two genotypes, with 21 down-regulated and 79 up-regulated in the ostst1 mutants (DOCX 139 KB)
425_2023_4160_MOESM9_ESM.docx
Supplementary file9 Heat maps showing differentially expressed genes (DEGs) between wild type (WT) and ostst1#3 plants at 10 d after flowering (DAF). DEGs were identified in flag leaves with a false discovery rate (FDR) threshold of < 0.01. There were 570 DEGs between the two genotypes, with 359 down-regulated and 211 up-regulated in the ostst1 mutants (DOCX 248 KB)
425_2023_4160_MOESM10_ESM.docx
Supplementary file10 Gene ontology (GO) enrichment analysis of differentially expressed genes (DEGs). DEGs were analyzed between wild-type (WT) and ostst1#3 leaves at 14 DAG (a) and at 10 d DAF (b) (DOCX 314 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
Yang, MY., Yang, X., Yan, Z. et al. OsTST1, a key tonoplast sugar transporter from source to sink, plays essential roles in affecting yields and height of rice (Oryza sativa L.). Planta 258, 4 (2023). https://doi.org/10.1007/s00425-023-04160-w
Received:
Accepted:
Published:
DOI: https://doi.org/10.1007/s00425-023-04160-w