Skip to main content
SpringerLink
Log in

Multiple Transgenic Strategies Positively Regulate Cold-Induced Sweetening in Low Temperature Stored Potato Tubers

  • Published:
Potato Research Aims and scope Submit manuscript

Abstract

Cold-induced sweetening (CIS) is a common phenomenon in many plants including potatoes that help in osmoregulation and cryoprotection. However, CIS is associated with quality deterioration in potato tubers due to accumulation of reducing sugars at low temperatures. We investigated two different strategies to modulate CIS in potato, overexpression of RING finger (SbRFP1) as anti-sweetening gene and by double-stranded RNA-mediated gene silencing of the vacuolar invertase gene. In silico analysis predicted that the ubiquitination activity of the RING finger protein was responsible for regulating the expression of invertase during cold-induced stress. Moreover, the in silico predicted binding stability of siRNA-mRNA duplex suggested efficient gene silencing of the invertase gene. We successfully generated four single and three dual transgenic potato lines that were positive for transgene insertion and integration as revealed in PCR and Southern blot. The amount of reducing sugars found in tubers obtained from single transgenics showed maximum decrease of 1.67 folds while tubers obtained from dual transgenic line depicted 4.86 folds reduced accumulation of reducing sugars compared to non-transgenic control when analyzed through HPLC analysis post 60-day storage at low temperature (4°C). Further, the invertase activity was 1.46 folds reduced in single transgenics while this reduction was 2.13 folds in dual transgenics. The downregulation of the invertase gene was up to 3.36 folds in dual transgenic potato lines, 2.26 folds in single transgenic compared to control, non-transgenic post 60-day cold storage at low temperature. Conclusively, the utilization of multiple strategies to regulate CIS in low-temperature stored potato tubers positively regulate CIS in transgenic potatoes and can be employed to generate CIS resistant potato varieties.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

Data Availability

No data associated in the manuscript.

References

  • Abbas G, Hafiz I, Abbasi N, Hussain A (2012) Determination of processing and nutritional quality attributes of potato genotypes in Pakistan. Pak J Bot 44:201–208

    Google Scholar 

  • Ali A, Iqbal M, Ali Q, Razzaq A, Nasir IA (2016) Gene profiling for invertase activity: assessment of potato varieties for resistance towards cold induced sweetening. Adv Life Sci 3(2):63–70

    CAS  Google Scholar 

  • 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 154:939–948

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Borda D, Alexe P (2011) Acrylamide levels in food. Romanian J Food Sci 1:3–15

    Google Scholar 

  • Bradshaw J, Bryan G, Ramsay G (2006) Genetic resources (including wild and cultivated Solanum species) and progress in their utilisation in potato breeding. Potato Res 49:49–65

    Article  Google Scholar 

  • Chakraborty S, Chakraborty N, Datta A (2000) Increased nutritive value of transgenic potato by expressing a nonallergenic seed albumin gene from Amaranthus hypochondriacus. Proc Natl Acad Sci 97:3724–3729

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Chan CY, Carmack CS, Long DD, Maliyekkel A, Shao Y, Roninson IB, Ding Y (2009) A structural interpretation of the effect of GC-content on efficiency of RNA interference. BMC Bioinform 10:1–7

    Article  Google Scholar 

  • Datir SS, Latimer JM, Thomson SJ, Ridgway HJ, Conner AJ, Jacobs JM (2012) Allele diversity for the apoplastic invertase inhibitor gene from potato. Mol Genet Genomics 287:451–460

    Article  CAS  PubMed  Google Scholar 

  • Dourado C, Pinto C, Barba FJ, Lorenzo JM, Delgadillo I, Saraiva JA (2019) Innovative non-thermal technologies affecting potato tuber and fried potato quality. Trends Food Sci Technol 88:274–289

    Article  CAS  Google Scholar 

  • Dove KK, Klevit RE (2017) RING-between-RING E3 ligases: emerging themes amid the variations. J Mol Biol 429:3363–3375

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Hannon GJ (2002) RNA interference. Nature 418:244–251

    Article  CAS  PubMed  Google Scholar 

  • Hatakeyama S, Nakayama KI (2003) Ubiquitylation as a quality control system for intracellular proteins. J Biochem 134:1–8

    Article  CAS  PubMed  Google Scholar 

  • He J, Wang J, Tao H, Xiao Y, Huang S-Y (2019) HNADOCK: a nucleic acid docking server for modeling RNA/DNA–RNA/DNA 3D complex structures. Nucleic Acids Res 47:W35–W42

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Hofacker IL (2003) Vienna RNA secondary structure server. Nucleic Acids Res 31:3429–3431

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Hou J, Zhang H, Liu J, Reid S, Liu T, Xu S, Tian Z, Sonnewald U, Song B, Xie C (2017) Amylases StAmy23, StBAM1 and StBAM9 regulate cold-induced sweetening of potato tubers in distinct ways. J Exp Bot 68:2317–2331

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Hsu Y, Goodman S (2005) Spectrin and ubiquitination: a review. Cell Mol Biol (Noisy-le-grand):OL801–807

  • Jaiswal S, Paul K, Raman KV, Tyagi S, Saakre M, Tilgam J, Bhattacharjee S, Vijayan J, Mondal KK, Sreevathsa R (2023) Amelioration of cold-induced sweetening in potato by RNAi mediated silencing of StUGPase encoding UDP-glucose pyrophosphorylase. Front Plant Sci 14:1133029

    Article  PubMed  PubMed Central  Google Scholar 

  • Joazeiro CA, Weissman AM (2000) RING finger proteins: mediators of ubiquitin ligase activity. Cell 102:549–552

    Article  CAS  PubMed  Google Scholar 

  • Kamrani M, BaghbanKohnehrouz B, Gholizadeh A (2016) Effect of RNAi-mediated gene silencing of starch phosphorylase L and starch-associated R1 on cold-induced sweetening in potato. J Hortic Sci Biotechnol 91:625–633

    Article  CAS  Google Scholar 

  • Kanehisa M, Goto S (2000) KEGG: Kyoto encyclopedia of genes and genomes. Nucleic Acids Res 28:27–30

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kumar D, Paul V, Ezekiel R (2005) Chipping quality of potatoes stored in heaps and pits in subtropical plains of India. Hort Sci (prague) 32:23–30

    Article  Google Scholar 

  • 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 67:41–47

    Article  CAS  PubMed  Google Scholar 

  • Li M, Xie C, Song B, Ou Y, Lin Y, Liu X, Zhang H, Liu J (2015) Construction of efficient, tuber-specific, and cold-inducible promoters in potato. Plant Sci 235:14–24

    Article  CAS  PubMed  Google Scholar 

  • Liu K, Wang L, Xu Y, Chen N, Ma Q, Li F, Chong K (2007) Overexpression of OsCOIN, a putative cold inducible zinc finger protein, increased tolerance to chilling, salt and drought, and enhanced proline level in rice. Planta 226:1007–1016

    Article  CAS  PubMed  Google Scholar 

  • Liu X, Zhang C, Ou Y, Lin Y, Song B, Xie C, Liu J, Li X-Q (2011) Systematic analysis of potato acid invertase genes reveals that a cold-responsive member, StvacINV1, regulates cold-induced sweetening of tubers. Mol Genet Genomics 286:109–118

    Article  CAS  PubMed  Google Scholar 

  • Liu X, Shi W, Yin W, Wang J (2017) Distinct cold responsiveness of a StInvInh2 gene promoter in transgenic potato tubers with contrasting resistance to cold-induced sweetening. Plant Physiol Biochem 111:77–84

    Article  CAS  PubMed  Google Scholar 

  • Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2− ΔΔCT method. Methods 25:402–408

    Article  CAS  PubMed  Google Scholar 

  • Matsuura-Endo C, Kobayashi A, Noda T, Takigawa S, Yamauchi H, Mori M (2004) Changes in sugar content and activity of vacuolar acid invertase during low-temperature storage of potato tubers from six Japanese cultivars. J Plant Res 117:131–137

    Article  CAS  PubMed  Google Scholar 

  • 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 82:231–239

    Article  CAS  Google Scholar 

  • 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 36:176–185

    Article  CAS  PubMed  Google Scholar 

  • Miki D, Shimamoto K (2004) Simple RNAi vectors for stable and transient suppression of gene function in rice. PCP 45:490–495

    CAS  Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Plant Physiol 15:473–497

    Article  CAS  Google Scholar 

  • Murtaza S, Tabassum B, Tariq M, Riaz S, Yousaf I, Jabbar B, Khan A, Samuel AO, Zameer M, Nasir IA (2022) Silencing a myzus persicae macrophage inhibitory factor by plant-mediated RNAi induces enhanced aphid mortality coupled with boosted RNAi efficacy in transgenic potato lines. Mol Biotechnol 64:1152–1163

    Article  CAS  PubMed  Google Scholar 

  • Ranjan R, Patro S, Pradhan B, Kumar A, Maiti IB, Dey N (2012) Development and functional analysis of novel genetic promoters using DNA shuffling, hybridization and a combination thereof. PLoS ONE 7:e31931

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Santner A, Estelle M (2010) The ubiquitin-proteasome system regulates plant hormone signaling. Plant J 61:1029–1040

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Sievers F, Higgins DG (2018) Clustal Omega for making accurate alignments of many protein sequences. Protein Sci 27:135–145

    Article  CAS  PubMed  Google Scholar 

  • Sowokinos JR (2001) Biochemical and molecular control of cold-induced sweetening in potatoes. Am J Potato Res 78:221–236

    Article  CAS  Google Scholar 

  • Sui G, Soohoo C, Affar EB, Gay F, Shi Y, Forrester WC, Shi Y (2002) A DNA vector-based RNAi technology to suppress gene expression in mammalian cells. Proc Natl Acad Sci 99:5515–5520

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Tabassum B, Nasir IA, Khan A, Aslam U, Tariq M, Shahid N, Husnain T (2016) Short hairpin RNA engineering: in planta gene silencing of potato virus Y. Crop Prot 86:1–8

    Article  CAS  Google Scholar 

  • Tareke E, Rydberg P, Karlsson P, Eriksson S, Törnqvist M (2002) Analysis of acrylamide, a carcinogen formed in heated foodstuffs. J Agric Food Chem 50:4998–5006

    Article  CAS  PubMed  Google Scholar 

  • Uchida C, Kitagawa M (2016) RING-, HECT-, and RBR-type E3 ubiquitin ligases: involvement in human cancer. Curr Cancer Drug Targets 16:157–174

    Article  CAS  PubMed  Google Scholar 

  • Verma P, Rao A, Singh PJ (2018) In silico prediction and designing of potential siRNA to control cotton whitefly Bemisia tabaci Gennadius vectored cotton leaf curl virus (CLCuV)

  • 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 7:1–10

    Article  Google Scholar 

  • Wu W, Cheng Z, Liu M, Yang X, Qiu D (2014) C3HC4-type RING finger protein Nb ZFP1 is involved in growth and fruit development in Nicotiana benthamiana. PLoS ONE 9:e99352

    Article  PubMed  PubMed Central  Google Scholar 

  • Zhang H, Liu X, Liu J, Ou Y, Lin Y, Li M, Song B, Xie C (2013) A novel RING finger gene, SbRFP1, increases resistance to cold-induced sweetening of potato tubers. FEBS Lett 587:749–755

    Article  CAS  PubMed  Google Scholar 

  • Zhang H, Yao Y, Chen S, Hou J, Yu Y, Liu T, Du J, Song B, Xie C (2019) SbRFP1 regulates cold-induced sweetening of potato tubers by inactivation of StBAM1. Plant Physiol Biochem 136:215–221

    Article  CAS  PubMed  Google Scholar 

  • Zrenner R, Schüler K, Sonnewald U (1996) Soluble acid invertase determines the hexose-to-sucrose ratio in cold-stored potato tubers. Planta 198:246–252

    Article  CAS  PubMed  Google Scholar 

Download references

Funding

This work was supported by the Higher Education Commission (HEC) Pakistan through NRPU grant # 4072.

Author information

Authors and Affiliations

  1. Centre of Excellence in Molecular Biology, University of the Punjab Lahore, Lahore, Pakistan

    Nida Toufiq & Bushra Tabassum

  2. Department of Chemistry, Physics and Atmospheric Sciences Jackson State University, Jackson, USA

    Olawale Samuel Adeyinka

  3. Department of Microbiology, BUITEMS, Quetta, Pakistan

    Anwar Khan

  4. Faculty of Agricultural Sciences, University of the Punjab, Lahore, Pakistan

    Shazia Shafique & Sobiya Shafique

  5. Department of Biotechnology, BUITEMS, Quetta, Pakistan

    Nusrat Jahan

  6. Forman Christian College, Lahore, Pakistan

    Muhammad Umar Bhatti

  7. School of Biological Sciences, University of the Punjab, Quaid-I-Azam Campus, Lahore, 54590, Pakistan

    Rida Khalid, Ayman Naeem, Qamar Abbas & Bushra Tabassum

Authors
  1. Nida Toufiq
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Olawale Samuel Adeyinka
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Anwar Khan
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Shazia Shafique
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Nusrat Jahan
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Muhammad Umar Bhatti
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Rida Khalid
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Ayman Naeem
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Qamar Abbas
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Sobiya Shafique
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Bushra Tabassum
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

NT: investigation, AOS: data curation, AK: methodology, SS: data curation, NJ: writing, MUB: validation, RK: statistical analysis, AN: in silico studies, QA: in silico studies, SS: writing, BT: conceptualization and supervision.

Corresponding author

Correspondence to Bushra Tabassum.

Ethics declarations

Ethical Approval

This article does not contain any studies with human participants or animals performed by any of the authors.

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.

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.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Toufiq, N., Adeyinka, O.S., Khan, A. et al. Multiple Transgenic Strategies Positively Regulate Cold-Induced Sweetening in Low Temperature Stored Potato Tubers. Potato Res. (2024). https://doi.org/10.1007/s11540-024-09733-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1007/s11540-024-09733-0

Keywords

Search

Navigation