农业生物技术学报2024，Vol.32Issue(7)：1693-1702,10.DOI:10.3969/j.issn.1674-7968.2024.07.020
利用CRISPR/Cas9技术创制黄果番茄新种质
Creating New Germplasm of Yellow Fruit Tomato(Solanum lycopersicum)Using CRISPR/Cas9 Technology
摘要
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.关键词
番茄/CRISPR/Cas9/八氢番茄红素合成酶1(PSY1)/基因编辑/黄果番茄Key words
Tomato/CRISPR/Cas9/Phytoene synthase1(PSY1)/Genome editing/Yellow fruit tomato分类
农业科技引用本文复制引用
龙海成,马燕勤,周玉洁,常伟,李菊,李志,苗明军,杨亮..利用CRISPR/Cas9技术创制黄果番茄新种质[J].农业生物技术学报,2024,32(7):1693-1702,10.基金项目
国家现代农业产业技术体系四川省蔬菜创新团队(蔬菜分子辅助育种岗位)项目[川农函(2019)472号] （蔬菜分子辅助育种岗位）
四川省科技计划项目(2021YFYZ0022 （）
2023NSFSC0162) （）
四川省财政自主创新专项项目(2022ZZCX050) （2022ZZCX050）
