The vacuolar membrane sucrose transporter MdSWEET16 plays essential roles in the cold tolerance of apple

  • Guanxian Yang
  • Haifeng Xu
  • Qi Zou
  • Jing Zhang
  • Shenghui Jiang
  • Hongcheng Fang
  • Yicheng Wang
  • Mengyu Su
  • Nan WangEmail author
  • Xuesen ChenEmail author
Original Article


Sugar content and cold tolerance are important apple (Malus × domestica) characteristics targeted by breeding programs. Here, a vacuolar membrane sucrose transporter, MdSWEET16, was cloned and characterized. A phylogenetic tree analysis found that MdSWEET16 was on the same polygenetic branch as Arabidopsis thaliana SWEET16 and SWEET17. MdSWEET16 was located on chromosome 2 and consists of six exons and five introns. The recombinant protein was obtained by prokaryotic induction, and the amino acid sequence and transmembrane domain were analyzed by bioinformatics. The encoded ~ 30-kDa protein has seven transmembrane domains and is localized on the tonoplasts of ‘Orin’ callus protoplasts. The expression of MdSWEET16 changed in response to sucrose and low temperature in ‘Orin’ calli. In addition, we also analyzed the expression level of MdSWEET16 during different fruit developmental stages using qRT-PCR. MdSWEET16 was highly expressed in young fruit, and its expression level during fruit development was significantly negatively correlated with sucrose content as assessed by a quantitative fluorescence analysis. Overexpression of MdSWEET16 in ‘Orin’ calli could reduce their sucrose content but increased their cold tolerance compared with MdSWEET16 RNA interference calli, which indicated that MdSWEET16 is involved in the sucrose transport and cold tolerance of apple.

Key message

MdSWEET16’s expression level was significantly correlated with the sucrose content and low temperature, and was induced by sucrose and low temperatures. Overexpression of MdSWEET16 increased ‘Orin’ calli cold tolerance. The prokaryotic expression of its recombinant protein can be used to further study the function of the MdSWEET16 protein in apple.


Apple Tonoplast Sucrose transporter SWEET16 Cold resistance 



Quantitative real-time PCR


Monosaccharide transporters


Non-monosaccharide transporter




2,4-Dichlorophenoxyacetic acid


Hexadecyl trimethyl ammonium bromide


Isopropyl β-d-thiogalactoside


Sodium dodecyl sulfate polyacrylamide gel electrophoresis


4-Morpholineethanesulfonic acid



This work was supported by Shandong Provincial Agricultural Variety Project (2019LZGC007), National Natural Science Foundation of China (31730080) and National Key Research and Development Project of China (2016YFC0501505). We thank Liwen Bianji, Edanz Group China (, for editing the English text of a draft of this manuscript.

Author contributions

XC and GY: conceived and designed the experiments. GY and HX: performed the experiments. GY: analyzed the data. QZ, JZ, SJ, HF, YW, MS: contributed reagents/materials/analysis tools. GY and XC: wrote the paper.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

11240_2019_1717_MOESM1_ESM.jpg (329 kb)
Figure S1. Primer list (JPG 329 kb).


  1. Aleksandra H, Nicolas S, Fernando C, Fernie AR, Bernhard G, Christina K (2006) Sucrose transporter LeSUT1 and LeSUT2 inhibition affects tomato fruit development in different ways. Plant J 45:180–192CrossRefGoogle Scholar
  2. Anne E, Stefan M, Silvia S, Thomas S, Winfriede W, Peters SW, Felix K, Sacha B, Enrico M, Schmidt UG (2006) Identification of a vacuolar sucrose transporter in barley and Arabidopsis mesophyll cells by a tonoplast proteomic approach. Plant Physiol 141:196–207CrossRefGoogle Scholar
  3. Bürkle L, Hibberd JM, Quick WP, Kühn C, Hirner B, Frommer WB (1998) The H+-sucrose cotransporter NtSUT1 is essential for sugar export from tobacco leaves. Plant Physiol 118:59–68PubMedPubMedCentralCrossRefGoogle Scholar
  4. Büttner M, Sauer N (2000) Monosaccharide transporters in plants: structure, function and physiology. Biochim Biophys Acta 1465:263–274PubMedCrossRefGoogle Scholar
  5. Cao H, Guo S, Xu Y, Jiang K, Jones AM, Chong K (2011) Reduced expression of a gene encoding a Golgi localized monosaccharide transporter (OsGMST1) confers hypersensitivity to salt in rice (Oryza sativa). J Exp Bot 62:4595–4604PubMedPubMedCentralCrossRefGoogle Scholar
  6. Chao-Yang F, Jia-Xuan H, Xiao-Xue H, Jing J (2015) Genome-wide identification, phylogeny, and expression analysis of the SWEET gene family in tomato. Gene 573:261–272CrossRefGoogle Scholar
  7. Chen X, Tao F, Zhang Y, He T, Feng J, Zhang C (2007) Genetic diversity of volatile components in Xinjiang wild apple (Malus sieversii). J Genet Genomics 34:171–179PubMedCrossRefGoogle Scholar
  8. Chen XS, Han MY, Gui-Lin SU, Liu FZ, Guo GN, Jiang YM, Mao ZQ, Peng FT, Shu HR (2010) Discussion on today’s world apple industry trends and the suggestions on sustainable and efficient development of apple industry in China. J Fruit Sci 27:598–604Google Scholar
  9. Chong J, Piron MC, Meyer S, Merdinoglu D, Bertsch C, Mestre P (2014) The SWEET family of sugar transporters in grapevine: VvSWEET4 is involved in the interaction with Botrytis cinerea. J Exp Bot 65:6589–6601PubMedPubMedCentralCrossRefGoogle Scholar
  10. Davies C, Wolf T, Robinson SP (1999) Three putative sucrose transporters are differentially expressed in grapevine tissues. Plant Sci 147:93–100CrossRefGoogle Scholar
  11. Divya C (2015) Co-option of developmentally regulated plant SWEET transporters for pathogen nutrition and abiotic stress tolerance. IUBMB Life 67:461–471CrossRefGoogle Scholar
  12. Du F, Shi H, Zhang X, Xu X (2013) Responses of reactive oxygen scavenging enzymes, proline and malondialdehyde to water deficits among six secondary successional seral species in Loess Plateau. PLoS ONE 9:e98872CrossRefGoogle Scholar
  13. Duan N, Bai Y, Sun H, Wang N, Ma Y, Li M, Wang X, Jiao C, Legall N, Mao L (2017) Genome re-sequencing reveals the history of apple and supports a two-stage model for fruit enlargement. Nat Commun 8:249PubMedPubMedCentralCrossRefGoogle Scholar
  14. Eom JS, Cho JI, Reinders A, Lee SW, Yoo Y, Tuan PQ, Choi SB, Bang G, Park YI, Cho MH (2011) Impaired function of the tonoplast-localized sucrose transporter in rice, OsSUT2, limits the transport of vacuolar reserve sucrose and affects plant growth. Plant Physiol 157:109–119PubMedPubMedCentralCrossRefGoogle Scholar
  15. Farajzadeh M, Rahimi M, Kamali GA, Mavrommatis T (2010) Modelling apple tree bud burst time and frost risk in Iran. Meteorol Appl 17:45–52Google Scholar
  16. Feng L, Frommer WB (2015) Structure and function of SemiSWEET and SWEET sugar transporters. Trends Biochem Sci 40:480–486PubMedPubMedCentralCrossRefGoogle Scholar
  17. Guo JM, Shi ZA, Wang C, Zhang YH, Yan WD, Yang C (2013) Capillary electrophoresis determination method of sugar accumulation in deciduous tree fruit. Exp Sci Technol 11:44–46Google Scholar
  18. Guo WJ, Nagy R, Chen HY, Pfrunder S, Yu YC, Santelia D, Frommer WB, Martinoia E (2014) SWEET17, a facilitative transporter, mediates fructose transport across the tonoplast of Arabidopsis roots and leaves. Plant Physiol 164:777PubMedPubMedCentralCrossRefGoogle Scholar
  19. Han L, Zhu Y, Liu M, Zhou Y, Lu G, Lan L, Wang X, Zhao Y, Zhang XC (2017) Molecular mechanism of substrate recognition and transport by the AtSWEET13 sugar transporter. Proc Natl Acad Sci USA 114:10089PubMedCrossRefGoogle Scholar
  20. Jian H, Lu K, Yang B, Wang T, Zhang L, Zhang A, Wang J, Liu L, Qu C, Li J (2016) Genome-wide analysis and expression profiling of the SUC and SWEET gene families of sucrose transporters in oilseed rape (Brassica napus L.). Front Plant Sci 7:1464PubMedPubMedCentralGoogle Scholar
  21. Kanno Y, Oikawa T, Chiba Y, Ishimaru Y, Shimizu T, Sano N, Koshiba T, Kamiya Y, Ueda M, Seo M (2016) AtSWEET13 and AtSWEET14 regulate gibberellin-mediated physiological processes. Nat Commun 7:13245PubMedPubMedCentralCrossRefGoogle Scholar
  22. Kiyosue T, Abe H, Yamaguchishinozaki K, Shinozaki K (1998) ERD6, a cDNA clone for an early dehydration-induced gene of Arabidopsis, encodes a putative sugar transporter. Biochem Biophys Acta 1370:187–191PubMedCrossRefGoogle Scholar
  23. Klappach G (2010) Biosynthesis of the major crop products. Acta Biotechnol 13:305CrossRefGoogle Scholar
  24. Klemens PAW, Patzke K, Deitmer J, Spinner L, Le Hir R, Bellini C, Bedu M, Chardon F, Krapp A, Neuhaus HE (2013) Overexpression of the vacuolar sugar carrier AtSWEET16 modifies germination, growth, and stress tolerance in Arabidopsis. Plant Physiol 163:1338–1352PubMedPubMedCentralCrossRefGoogle Scholar
  25. Liang Y, Wang K, Xue Q, Sui L, Ye J, Chen X (2017) Effects of exogenous growth substances on physiological traits of cold tolerance in Citrus aurantium seedlings. Sci Silvae Sin 53:68–75Google Scholar
  26. Li-Qing C (2014) SWEET sugar transporters for phloem transport and pathogen nutrition. New Phytol 201:1150–1155CrossRefGoogle Scholar
  27. Li-Qing C, Bi-Huei H, Sylvie L, Hitomi T, Hartung ML, Xiao-Qing Q, Woei-Jiun G, Jung-Gun K, William U, Bhavna C (2010) Sugar transporters for intercellular exchange and nutrition of pathogens. Nature 468:527–532CrossRefGoogle Scholar
  28. Li-Qing C, Xiao-Qing Q, Bi-Huei H, Davide S, Sonia O, Fernie AR, Frommer WB (2012) Sucrose efflux mediated by SWEET proteins as a key step for phloem transport. Science 335:207CrossRefGoogle Scholar
  29. Liu X, Zhang Y, Yang C, Tian Z, Li J (2016) AtSWEET4, a hexose facilitator, mediates sugar transport to axial sinks and affects plant development. Sci Rep UK 6:24563CrossRefGoogle Scholar
  30. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 25:402–408Google Scholar
  31. Ma XL, Liu YH, Yuan ZL, Shi YS, Song YC, Wang TY, Yu L (2009) Cloning of cDNAs for a novel sugar transporter gene, ZmERD6, from maize and its expression analysis under abiotic stresses. Acta Agron Sin 35:1410–1417CrossRefGoogle Scholar
  32. Mahajan S, Tuteja N (2005) Cold, salinity and drought stresses: an overview. Arch Biochem Biophys 444:139–158PubMedPubMedCentralCrossRefGoogle Scholar
  33. Martinoia E, Meyer S, Angeli AD, Nagy R (2012) Vacuolar transporters in their physiological context. Annu Rev Plant Biol 63:183PubMedCrossRefGoogle Scholar
  34. Miao H, Sun P, Liu Q, Miao Y, Liu J, Zhang K, Hu W, Zhang J, Wang J, Wang Z (2017) Genome-wide analyses of SWEET family proteins reveal involvement in fruit development and abiotic/biotic stress responses in banana. Sci Rep 7:3536PubMedPubMedCentralCrossRefGoogle Scholar
  35. Michael W, John N, Carole B, Timothy A, Dumitru M (2011) Ectopic expression of a novel peach (Prunus persica) CBF transcription factor in apple (Malus × domestica) results in short-day induced dormancy and increased cold hardiness. Planta 233:971–983CrossRefGoogle Scholar
  36. Ming-Hui LU, Song H, Xiao-Ming LI, Chen JF (2005) Changes of SOD, CAT and POD activities in cucumber leaves during cold damage. Acta Bot Boreal Occident Sin 25:1570–1573Google Scholar
  37. Rae AL, Perroux JM, Grof CPL (2005) Sucrose partitioning between vascular bundles and storage parenchyma in the sugarcane stem: a potential role for the ShSUT1 sucrose transporter. Planta 220:817–825PubMedCrossRefGoogle Scholar
  38. Ruan YL (2014) Sucrose metabolism: gateway to diverse carbon use and sugar signaling. Annu Rev Plant Biol 65:33–67PubMedCrossRefGoogle Scholar
  39. Sauer N, Ludwig A, Knoblauch A, Rothe P, Gahrtz M, Klebl F (2004) AtSUC8 and AtSUC9 encode functional sucrose transporters, but the closely related AtSUC6 and AtSUC7 genes encode aberrant proteins in different Arabidopsis ecotypes. Plant J 40:120–130PubMedCrossRefGoogle Scholar
  40. Schneider S, Hulpke S, Schulz A, Yaron I, Höll J, Imlau A, Schmitt B, Batz S, Wolf S, Hedrich R (2012) Vacuoles release sucrose via tonoplast-localised SUC4-type transporters. Plant Biol 14:325–336PubMedCrossRefGoogle Scholar
  41. Schulz A, Beyhl D, Marten I, Wormit A, Neuhaus E, Poschet G, Büttner M, Schneider S, Sauer N, Hedrich R (2011) Proton-driven sucrose symport and antiport are provided by the vacuolar transporters SUC4 and TMT1/2. Plant J 68:129–136PubMedCrossRefGoogle Scholar
  42. Seo PJ, Park JM, Kang SK, Kim SG, Park CM (2011) An Arabidopsis senescence-associated protein SAG29 regulates cell viability under high salinity. Planta 233:189–200CrossRefGoogle Scholar
  43. Stadler RE, Gahrtz M, Sauer N (2010) The AtSUC1 sucrose carrier may represent the osmotic driving force for anther dehiscence and pollen tube growth in Arabidopsis. PLANT J 19:269–278CrossRefGoogle Scholar
  44. Viswanathan C, Jianhua Z, Jian-Kang Z (2007) Cold stress regulation of gene expression in plants. Trends Plant Sci 12:444–451CrossRefGoogle Scholar
  45. Wang L, Yao L, Hao X, Li N, Qian W, Yue C, Ding C, Zeng J, Yang Y, Wang X (2018) Tea plant SWEET transporters: expression profiling, sugar transport, and the involvement of CsSWEET16 in modifying cold tolerance in Arabidopsis. Plant Mol Biol 96:577–592PubMedCrossRefGoogle Scholar
  46. Weigl K, Flachowsky H, Peil A, Hanke MV (2015) Heat mediated silencing of MdTFL1 genes in apple (Malus × domestica). Plant Cell Tissue Organ Cult 123:511–521CrossRefGoogle Scholar
  47. Wickett NJ, Siavash M, Nam N, Tandy W, Eric C, Naim M, Saravanaraj A, Barker MS, Burleigh JG, Gitzendanner MA (2014) Phylotranscriptomic analysis of the origin and early diversification of land plants. Proc Natl Acad Sci USA 111:4859–4868CrossRefGoogle Scholar
  48. Wippel K, Sauer N (2012) Arabidopsis SUC1 loads the phloem in suc2 mutants when expressed from the SUC2 promoter. J Exp Bot 63:669–679PubMedCrossRefGoogle Scholar
  49. Xu H, Wang N, Wang Y, Jiang S, Fang H, Zhang J, Su M, Zuo W, Xu L, Zhang Z (2018) Overexpression of the transcription factor MdbHLH33 increases cold tolerance of transgenic apple callus. Plant Cell Tissue Organ Cult 134:1–10CrossRefGoogle Scholar
  50. Zhao L, Yao J, Wei C, Yan LI, Youjun LÜ, Yan G, Wang J, Li Y, Liu Z, Zhang Y (2018) A genome-wide analysis of SWEET gene family in cotton and their expressions under different stresses. J Cotton Res 1:7CrossRefGoogle Scholar

Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  • Guanxian Yang
    • 1
  • Haifeng Xu
    • 1
  • Qi Zou
    • 1
  • Jing Zhang
    • 1
  • Shenghui Jiang
    • 1
  • Hongcheng Fang
    • 1
  • Yicheng Wang
    • 1
  • Mengyu Su
    • 1
  • Nan Wang
    • 1
    Email author
  • Xuesen Chen
    • 1
    Email author
  1. 1.National Key Laboratory of Crop Biology, College of Horticulture ScienceShandong Agricultural UniversityTai-AnChina

Personalised recommendations