Plant Biotechnology Reports

, Volume 12, Issue 3, pp 175–185 | Cite as

RNAi-mediated silencing of endogenous Vlnv gene confers stable reduction of cold-induced sweetening in potato (Solanum tuberosum L. cv. Désirée)

  • Amir Hameed
  • Rakhshanda Bilal
  • Farooq Latif
  • Joyce Van Eck
  • Georg Jander
  • Shahid Mansoor
Original Article


Potato tubers must be cold-stored to extend their shelf life and maintain an uninterrupted supply chain for food processors. However, a side-effect of low-temperature storage is manifested in terms of cold-induced sweetening (CIS) of potato tubers, which reduces the processing quality and the commercial value of the end-products. RNA interference (RNAi) technology, whereby transgene-derived small interfering RNAs can trigger the homology-based knockdown of cognate host genes and can initiate gene silencing, has been successfully applied in crop improvement through targeted gene knockout in host plants. In the current study, transgenic potato plants (Solanum tuberosum cv. Désirée) were generated, expressing a 300 bp hairpin loop nucleotide sequence targeting the potato vacuolar invertase gene (VInv), under the constitutive Cauliflower mosaic virus 35S promoter. Tubers collected from transgenic lines showed a significant reduction in reducing sugar content after 180 days of cold storage, without showing any measurable off-target effects on plant morphology and tuberization compared to non-transformed control plants. The cold-stored tubers were further assayed for chip color, which showed a fairly light colored quality in the samples originating from RNAi lines. Together with similar effects seen in previously published experiments involving other potato varieties, the Désirée results described here establish the efficacy of using RNAi for the successful reduction of CIS in potato tubers.


Cold storage RNAi Cold induced sweetening Vacuolar invertase Potato 



This work was funded by Higher Education Commission (HEC), Government of Pakistan, under the indigenous Ph.D. fellowship and research initiative program (IRSIP), and United States Department of Agriculture award 2014-67013-21659 to GJ. The authors would like to thank Ms. Kerry Swartwood for the assistance in tissue culture work that was conducted at the Boyce Thomason Institute for Plant Research.

Author contributions

AH conducted the research work and prepared the first draft of the manuscript. RB conceived the idea and proposed a layout for research work. FL provided the assistance for sugar content profiling using HPLC. JVE and GJ provided the assistance for the tissue culture work and molecular confirmation of transgenics. SM supervised the whole study and provided workspace and assistance during the entire study. The final draft of the manuscript was edited and approved by all co-authors.

Compliance with ethical standards

Conflict of interest

The authors declare no financial or commercial conflict of interest.


  1. Ausubel FM et al. (2002) Short protocols in molecular biology: a compendium of methods from current protocols in molecular biology. Wiley, New YorkGoogle Scholar
  2. Baghban-Kohnehrouz B, Nayeri S (2016) Design, simplified cloning, and in-silico analysis of multisite small interfering RNA-targeting cassettes. Mol Biol Res Commun 5:31PubMedPubMedCentralGoogle Scholar
  3. Barrell PJ, Meiyalaghan S, Jacobs JM, Conner AJ (2013) Applications of biotechnology and genomics in potato improvement. Plant Biotechnol J 11:907–920CrossRefPubMedGoogle Scholar
  4. Bethke PC, Sabba R, Bussan AJ (2009) Tuber water and pressure potentials decrease and sucrose contents increase in response to moderate drought and heat stress. Am J Potato Res 86:519–532CrossRefGoogle Scholar
  5. Bhaskar PB, Wu L, Busse JS, Whitty BR, Hamernik AJ, Jansky SH, Buell CR, Bethke PC, Jiang J (2010) Suppression of the vacuolar invertase gene prevents cold-induced sweetening in potato. Plant Physiol 154:939–948CrossRefPubMedPubMedCentralGoogle Scholar
  6. Blenkinsop R, Yada R, Marangoni A (2004) Metabolic control of low-temperature sweetening in potato tubers during postharvest storage. Hortic Rev 30:317–354Google Scholar
  7. Chang CI, Yoo JW, Hong SW, Lee SE, Kang HS, Sun X, Rogoff HA, Ban C, Kim S, Li CJ (2009) Asymmetric shorter-duplex siRNA structures trigger efficient gene silencing with reduced nonspecific effects. Mol Ther 17:725–732CrossRefPubMedPubMedCentralGoogle Scholar
  8. Cheng S, Liu J, Xie C, Song B, Liu X, Li J (2006) Role of tobacco vacuolar invertase regulated by patatin promoter in resistance of potato tubers to cold-sweetening. J Agric Biotech 14:716–720Google Scholar
  9. Chi Z, Cong-Hua X, Bo-Tao S, Xun L, Jun L (2008) RNAi effects on regulation of endogenous acid invertase activity in potato (Solanum tuberosum L.) tubers. Chin J Agric Biotech 5:107–112CrossRefGoogle Scholar
  10. Dale MFB, Bradshaw JE (2003) Progress in improving processing attributes in potato. Trends in Plant Sci 8:310–312CrossRefGoogle Scholar
  11. Friedman M (2003) Chemistry, biochemistry, and safety of acrylamide, a review. J Agric Food Chem 51:4504–4526CrossRefPubMedGoogle Scholar
  12. Greiner S, Rausch T, Sonnewald U, Herbers K (1999) Ectopic expression of a tobacco invertase inhibitor homolog prevents cold-induced sweetening of potato tubers. Nat Biotech 17:708–711CrossRefGoogle Scholar
  13. Hameed A, Tahir MN, Asad S, Bilal R, Van Eck J, Jander G, Mansoor S (2017) RNAi-mediated simultaneous resistance against three RNA viruses in potato. Mol Biotech 59:73–83CrossRefGoogle Scholar
  14. Hamernik A, Hanneman R, Jansky S (2009) Introgression of wild species germplasm with extreme resistance to cold sweetening into the cultivated potato. Crop Sci 49:529–542CrossRefGoogle Scholar
  15. Jacobs JM, Takla MF, Docherty LC, Frater CM, Markwick NP, Meiyalaghan S, Conner AJ (2009) Potato transformation with modified nucleotide sequences of the cry9Aa2 gene improves resistance to potato tuber moth. Potato Res 52:367–378CrossRefGoogle Scholar
  16. Keijbets M (2008) Potato processing for the consumer: developments and future challenges. Potato Res 51:271–281CrossRefGoogle Scholar
  17. Klann EM, Hall B, Bennett AB (1996) Antisense acid invertase (TIV1) gene alters soluble sugar composition and size in transgenic tomato fruit. Plant Physiol 112:1321–1330CrossRefPubMedPubMedCentralGoogle Scholar
  18. Li XQ (2008) Molecular characterization and biotechnological improvement of the processing quality of potatoes. Can J Plant Sci 88:639–648CrossRefGoogle Scholar
  19. Li M, Song B, Zhang Q, Liu X, Lin Y, Ou Y, Zhang H, Liu J (2013) A synthetic tuber-specific and cold-induced promoter is applicable in controlling potato cold-induced sweetening. Plant Physiol Biochem 67:41–47CrossRefPubMedGoogle Scholar
  20. Lindsay H (1973) A colorimetric estimation of reducing sugars in potatoes with 3, 5-dinitrosalicylic acid. Potato Res 16:176–179CrossRefGoogle Scholar
  21. Liu X, Lin Y, Liu J, Song B, Ou Y, Zhang H, Li M, Xie C (2013) StInvInh2 as an inhibitor of StvacINV1 regulates the cold-induced sweetening of potato tubers by specifically capping vacuolar invertase activity. Plant Biotechnol J 11:640–647CrossRefPubMedGoogle Scholar
  22. McCann LC, Bethke PC, Simon PW (2010) Extensive variation in fried chip color and tuber composition in cold-stored tubers of wild potato (Solanum) germplasm. J Agric Food Chem 58:2368–2376CrossRefPubMedGoogle Scholar
  23. McKenzie MJ, Sowokinos JR, Shea IM, Gupta SK, Lindlauf RR, Anderson JA (2005) Investigations on the role of acid invertase and UDP-glucose pyrophosphorylase in potato clones with varying resistance to cold-induced sweetening. Am J Potato Res 82:231–239CrossRefGoogle Scholar
  24. Mckenzie MJ, Chen RK, Harris JC, Ashworth MJ, Brummell DA (2013) Post-translational regulation of acid invertase activity by vacuolar invertase inhibitor affects resistance to cold-induced sweetening of potato tubers. Plant Cell Environ 36:176–185CrossRefPubMedGoogle Scholar
  25. Nägele T, Henkel S, Hörmiller I, Sauter T, Sawodny O, Ederer M, Heyer AG (2010) Mathematical modeling of the central carbohydrate metabolism in Arabidopsis reveals a substantial regulatory influence of vacuolar invertase on whole plant carbon metabolism. Plant Physiol 153:260–272CrossRefPubMedPubMedCentralGoogle Scholar
  26. Parzefall W (2008) Minireview on the toxicity of dietary acrylamide. Food Chem Toxi 46:1360–1364CrossRefGoogle Scholar
  27. Sambrook J, Frisch EF, Maniatis T (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, New YorkGoogle Scholar
  28. Stadler RH, Blank I, Varga N, Robert F, Hau J, Guy PA, Robert MC, Riediker S (2002) Food chemistry: acrylamide from Maillard reaction products. Nature 419:449–450CrossRefPubMedPubMedCentralGoogle Scholar
  29. Webb KJ, Osifo EO, Henshaw GG (1983) Shoot regeneration from leaflet discs of six cultivars of potato (Solanum tuberosum subsp. tuberosum). Plant Sci Lett 30:1–8CrossRefGoogle Scholar
  30. Wiberley-Bradford AE, Busse JS, Jiang J, Bethke PC (2014) Sugar metabolism, chip color, invertase activity, and gene expression during long-term cold storage of potato (Solanum tuberosum) tubers from wild-type and vacuolar invertase silencing lines of Katahdin. BMC Res Notes 7:801CrossRefPubMedPubMedCentralGoogle Scholar
  31. Xiong X, Tai G, Seabrook J, Wehling P (2002) Effectiveness of selection for quality traits during the early stage in the potato breeding population. Plant Breed 121:441–444CrossRefGoogle Scholar
  32. Yu X, Wang X, Zhang W, Qian T, Tang G, Guo Y, Zheng C (2008) Antisense suppression of an acid invertase gene (MAI1) in muskmelon alters plant growth and fruit development. J Exp Bot 59:2969–2977CrossRefPubMedGoogle Scholar
  33. Zhu X, Gong H, He Q, Zeng Z, Busse JS, Jin W, Bethke PC, Jiang J (2016) Silencing of vacuolar invertase and asparagine synthetase genes and its impact on acrylamide formation of fried potato products. Plant Biotech J 14:709–718CrossRefGoogle Scholar
  34. Zrenner R, Schüler K, Sonnewald U (1996) Soluble acid invertase determines the hexose-to-sucrose ratio in cold-stored potato tubers. Planta 198:246–252CrossRefPubMedGoogle Scholar

Copyright information

© Korean Society for Plant Biotechnology and Springer Japan KK, part of Springer Nature 2018

Authors and Affiliations

  • Amir Hameed
    • 1
    • 2
  • Rakhshanda Bilal
    • 1
  • Farooq Latif
    • 1
  • Joyce Van Eck
    • 3
  • Georg Jander
    • 3
  • Shahid Mansoor
    • 1
  1. 1.Molecular Virology and Gene Silencing GroupNational Institute for Biotechnology and Genetic EngineeringFaisalabadPakistan
  2. 2.Department of Bioinformatics and BiotechnologyGovernment College UniversityFaisalabadPakistan
  3. 3.Boyce Thompson Institute for Plant ResearchIthacaUSA

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