Vietnam Academy of Science and Technology

Project's information

Project's title	Enhancing the levels of sugar and amino acid by CRISPR/Cas9-mediated mutagenesis in tomatoes (Solanum lycopersicum)
Project’s code	ĐLTE00.10/20-21
Research hosting institution	Institute of Biotechnology
Project leader’s name	Dr. Do Tien Phat
Project duration	01/01/2020 - 31/12/2021
Project’s budget	500 million VND
Classify	Grade A
Goal and objectives of the project	The main goal of the project is to develop and utilize CRISPR/Cas9 system for generating potential mutant tomato lines with the enhanced accumulation of sugar and amino acid in fruits. Specific objectives: - Establish a sufficient method to create targeted mutations in Vietnamese tomato cultivar using the CRISPR/Cas9 system. - Generate potential tomtato lines with homozygous mutations in uORF regions of the SlbZIP1 gene.
Main results	- Scientific significance: This is the first success in utilizing the CRISPR/Cas9 system to induce targeted mutations in uORF regions of the SlbZIP, a transcription factor family, for improving tomato fruit quality. This result provides potential strategies for regulating bZIP expression via alteration of uORF sequences to enhance fruit quality of other plants. This is also the first study in genome editing of Vietnamese tomato cultivar using the CRISPR/Cas9 system and show potential for tomato breeding. - Applications: Tomato is one of the world's most consumed vegetable due to its high nutritional values. Fresh ripe fruits could be directly used as raw vegetables or materials for food processing. Therefore, the enhanced accumulation of sugar and amino acid contents will increase tomato fruit flavor and quality, consequently improve consumer’s health. These potential mutant lines also will be used for further tomato breeding and cultivation programs.
Novelty and actuality and scientific meaningfulness of the results	This is the first success in utilizing the CRISPR/Cas9 system to induce targeted mutations on the SlbZIP-uORF regions in tomato. This study demonstrated the correlation of the SlbZIP1 expression with the accumulation of sugar and amino acids in tomato mutant fruits. This result indicates that controlling SlbZIP1 expression via uORFs-targeted mutagenesis should be potential for enhancing tomato quality without impairing on plant growth and development.
Products of the project	- Publications supported by the project: Nguyen Doai Van, Hoang Trang Thi Huyen, Le Ngoc Thu, Tran Huyen Thi, Nguyen Cuong Xuan, Moon Yong-Hwan, Chu Ha Hoang, Do Phat Tien, 2022, “An Efficient Hairy Root System for Validation of Plant Transformation Vector and CRISPR/Cas Construct Activities in Cucumber (Cucumis sativus L.)”, Frontiers in Plant Science, Vol 12, DOI=10.3389/fpls.2021.770062 Nhung Hong Nguyen, Thao Phuong Bui, Ngoc Thu Le, Cuong Xuan Nguyen, My Tra Thi Le, Nhan Trong Dao, Quyen Phan, Trong Van Le, Huong Mai Thi To, Ngoc Bich Pham, Ha Hoang Chu, Phat Tien Do (2023), Disrupting Sc-uORFs of a transcription factor bZIP1 using CRISPR/Cas9 enhances sugar and amino acid contents in tomato (Solanum lycopersicum). Planta 257(3):57. doi: 10.1007/s00425-023-04089-0. PMID: 36795295. - Technological products: + Three mutant lines harboring the mutation in uORF region of SlbZIP1 which enhancing sugar and amino acid contents in tomatoes more than its wild-type; + A method in utilizing the CRISPR/Cas9 system for sufficiently induce targeted mutations in tomato.
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