Nida Toufiq, Olawale Samuel Adeyinka, Anwar Khan, Shazia Shafique, Nusrat Jahan, Muhammad Umar Bhatti, Rida Khalid, Ayman Naeem, Qamar Abbas, Sobiya Shafique, Bushra Tabassum
{"title":"Multiple Transgenic Strategies Positively Regulate Cold-Induced Sweetening in Low Temperature Stored Potato Tubers","authors":"Nida Toufiq, Olawale Samuel Adeyinka, Anwar Khan, Shazia Shafique, Nusrat Jahan, Muhammad Umar Bhatti, Rida Khalid, Ayman Naeem, Qamar Abbas, Sobiya Shafique, Bushra Tabassum","doi":"10.1007/s11540-024-09733-0","DOIUrl":null,"url":null,"abstract":"<p>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 (<i>SbRFP1</i>) 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.</p>","PeriodicalId":20378,"journal":{"name":"Potato Research","volume":"26 1","pages":""},"PeriodicalIF":2.3000,"publicationDate":"2024-05-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Potato Research","FirstCategoryId":"97","ListUrlMain":"https://doi.org/10.1007/s11540-024-09733-0","RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"AGRONOMY","Score":null,"Total":0}
引用次数: 0
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.
期刊介绍:
Potato Research, the journal of the European Association for Potato Research (EAPR), promotes the exchange of information on all aspects of this fast-evolving global industry. It offers the latest developments in innovative research to scientists active in potato research. The journal includes authoritative coverage of new scientific developments, publishing original research and review papers on such topics as:
Molecular sciences;
Breeding;
Physiology;
Pathology;
Nematology;
Virology;
Agronomy;
Engineering and Utilization.