CRISPR/cas9 based genetically edited potatoes with enhanced attributes | [Abstract Book on Second International Conference on Recent Approaches in Plant Sciences (RAPS-23) 4-5 May 2023 • 2023]

Author(s):

1. Aneela Yasmeen: Centre of Excellence in Molecular Biology, University of the Punjab, 87-West Canal Bank Road Lahore-53700, Pakistan

2. Abdul Qayyum Rao: Centre of Excellence in Molecular Biology, University of the Punjab, 87-West Canal Bank Road Lahore-53700, Pakistan

3. Saira Azam: Centre of Excellence in Molecular Biology, University of the Punjab, 87-West Canal Bank Road Lahore-53700, Pakistan

4. Ayesha Latif: Centre of Excellence in Molecular Biology, University of the Punjab, 87-West Canal Bank Road Lahore-53700, Pakistan

5. Naila Shahid: Centre of Excellence in Molecular Biology, University of the Punjab, 87-West Canal Bank Road Lahore-53700, Pakistan

6. Tahir Rehman Samiullah: Centre of Excellence in Molecular Biology, University of the Punjab, 87-West Canal Bank Road Lahore-53700, Pakistan

7. Allah Bakhsh: Centre of Excellence in Molecular Biology, University of the Punjab, 87-West Canal Bank Road Lahore-53700, Pakistan

8. Sana Shakoor: Centre of Excellence in Molecular Biology, University of the Punjab, 87-West Canal Bank Road Lahore-53700, Pakistan

9. Sara Ajmal: Centre of Excellence in Molecular Biology, University of the Punjab, 87-West Canal Bank Road Lahore-53700, Pakistan

10. Sahar Sadaqat: Centre of Excellence in Molecular Biology, University of the Punjab, 87-West Canal Bank Road Lahore-53700, Pakistan

11. Ayesha Imran: Centre of Excellence in Molecular Biology, University of the Punjab, 87-West Canal Bank Road Lahore-53700, Pakistan

12. Sania Naeem: Centre of Excellence in Molecular Biology, University of the Punjab, 87-West Canal Bank Road Lahore-53700, Pakistan

13. Muhammad Awais: Centre of Excellence in Molecular Biology, University of the Punjab, 87-West Canal Bank Road Lahore-53700, Pakistan

14. Muhammad Saad Bhutta: Centre of Excellence in Molecular Biology, University of the Punjab, 87-West Canal Bank Road Lahore-53700, Pakistan

15. Tayyab Husnain: Centre of Excellence in Molecular Biology, University of the Punjab, 87-West Canal Bank Road Lahore-53700, Pakistan

16. Kausar Malik: Centre of Excellence in Molecular Biology, University of the Punjab, 87-West Canal Bank Road Lahore-53700, Pakistan

Abstract:

Potato is the 4th largest food being known as complete food with enhanced nutritional value. Potato harvesting is usually done once in a year. To extend post harvesting and reduce sprouting cold storage of potatoes is commonly used. Unfortunately, when these potatoes are turned into potato chips or French fries at higher temperatures it results in production of a carcinogen i.e. acrylamide. Serious global food safety and concerns have been caused by prevalence of acrylamide in potato chips and French fries. CRISPR/ Cas9, the state of art technology was applied to counter this issue. Two sgRNAs were designed using CRISPR/direct online tool and plasmid was synthesized by using gateway cloning which was then transformed into a local cultivar of potato plants via Agrobacterium mediated transformation. The transformation efficiency was found to be 11.7%. Molecular analysis including PCR, sanger sequencing digital PCR ELISA were performed. The overall editing efficacy was determined to be 25.6% as per TIDE analysis. The amplicon sequencing data showed maximum indel frequency for potato plant T12 (14.3%) resulting in 6.2% gene knockout and 6% frame shift. While for plant B4, the maximum indel frequency of 2.0% was found which resulted in 4.4% knockout and 4% frameshift as analysed by Geneious. A reduction of 90-99% in mRNA expression of VInv gene was observed through qRT-PCR data analysis as compared to the non-edited control potato plant. Following cold storage, chips analysis of potatoes for reducing sugars’ analysis by titration method determined five-fold reduction in percentage of reducing sugars in tubers of B4 transgenic lines as compared to the control.

Page(s): 164-164

DOI: DOI not available

Published: Journal: Abstract Book on Second International Conference on Recent Approaches in Plant Sciences (RAPS-23) 4-5 May 2023 , Volume: 0, Issue: 0, Year: 2023

Keywords:

Reducing sugars , genome editing , Cold induced , CRISPRCas , Potatoes

References:

References are not available for this document.

Citations

Citations are not available for this document.

Citations

Downloads

Views