Creating New Germplasm of Yellow Fruit Tomato (Solanum lycopersicum) Using CRISPR/Cas9 Technology
LONG Hai-Cheng1,4, MA Yan-Qin1,2, ZHOU Yu-Jie1, CHANG Wei2,3, LI Ju1,2, LI Zhi1,2, MIAO Ming-Jun1,2, YANG Liang1,2,*
1 Vegetable Germplasm Innovation and Variety Improvement Key Laboratory of Sichuan Province/MOA of Key Laboratory of Horticultural Crops Biology and Germplasm Enhancement in Southwest Regions, Horticulture Research Institute, Sichuan Academy of Agricultural Sciences,Chengdu 610066, China; 2 Sichuan Province Engineering Technology Research Center of Vegetables, Pengzhou, Sichuan 611934, China; 3 Sichuan Institute of Edible Fungi, Chengdu 610066, China; 4 College of Resources, Sichuan Agricultural University, Chengdu 611130, China
Abstract At present, excellent yellow fruit tomato (Solanum lycopersicum) varieties are relatively scarce in the domestic market. In order to meet the market demand and improve the competitiveness of the seed industry, the tomato variety 'MoneyMaker' ('MM') was used to create yellow fruit by targeted gene editing of the tomato phytoene synthase 1 (SlPSY1) using CRISPR/Cas9 technology. Two sequences specific sgRNAs in the exon region of the SlPSY1 were selected as target sites to construct the CRISPR/Cas9 binary expression vector pKSE401. The 'MM' tomato material was genetically transformed using Agrobacterium tumefaciens mediated leaf disc transformation method. The results showed that 12 transgenic-positive plants were detected in 16 regenerated seedlings, with a positive transformation rate of 75%; Sequencing analysis of the target sites of the positive transgenic plants showed that 8 plants had a total of 21 editing events at 2 target sites, with a gene editing efficiency of 66.7%, and none of them was off-target; Five of the edited plants showed yellow fruit phenotype at maturity. In this study, CRISPR/Cas9 technology was used to successfully created SlPSY1 gene editing plants in 'MM' tomato, which will provide a technical and material basis for the rapid creation of tomato yellow fruit resources in the future.
LONG Hai-Cheng,MA Yan-Qin,ZHOU Yu-Jie, et al. Creating New Germplasm of Yellow Fruit Tomato (Solanum lycopersicum) Using CRISPR/Cas9 Technology[J]. 农业生物技术学报, 2024, 32(7): 1693-1702.
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