【摘要】 目前国内市场上优异黄果番茄(Solanum lycopersicum)品种相对稀缺，为了满足市场需求，提高种业竞争力，本研究以’MoneyMaker’(’MM’)番茄为材料，利用CRISPR/Cas9技术，对番茄八氢番茄红素合成酶1 (phytoene synthase 1, SlPSY1)基因(Solyc03g031860)进行定向基因编辑来创制黄色果实番茄。选择SlPSY1基因外显子区域2个序列特异的sgRNA作为靶位点，构建CRISPR/Cas9双元表达载体p KSE401，通过根癌农杆菌(Agrobacterium tumefaciens)介导的叶盘转化法对’MM’番茄材料进行遗传转化。结果显示，16株再生苗中鉴定出了12株阳性转基因植株，阳性转化率为75%；对阳性转基因植株的靶位点测序分析发现，有8株在2个靶位点上共发生21次编辑事件，基因编辑效率为66.7%，且均未发生脱靶现象；获得的突变植株中有5株在成熟期果实表现为黄色。本研究利用CRISPR/Cas9技术在’MM’番茄上成功创制了SlPSY1基因编辑材料，为今后快速创制番茄黄果资源提供技术及材料基础。更多还原

【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 SlPSY1gene 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.更多还原