Van Eck Joyce, Keen Patricia, Tjahjadi Michelle
The Boyce Thompson Institute, Ithaca, NY, USA.
Plant Breeding and Genetics Section, School of Integrative Plant Science, Cornell University, Ithaca, NY, USA.
Methods Mol Biol. 2019;1864:225-234. doi: 10.1007/978-1-4939-8778-8_16.
Tomato is both an important food crop and serves as a model plant species that is used for various research investigations including understanding gene function. Transformation is commonly utilized to facilitate these investigations in combination with all the extensive genetic and genomic resources available for tomato. The transformation protocol routinely used in our laboratory has been applied to many different tomato genotypes and relies on Agrobacterium tumefaciens infection of young cotyledon sections. We have used vector systems for overexpression, RNA interference for gene silencing, and CRISPR/Cas9 for genome editing. Vectors used to design gene constructs contained selectable marker genes that conferred resistance to kanamycin, hygromycin, and the herbicide component, bialaphos. The protocol we follow for Agrobacterium-mediated transformation of both cultivated and wild species of tomato is detailed in this chapter.
番茄既是一种重要的粮食作物，也是一种模式植物物种，可用于包括了解基因功能在内的各种研究调查。结合番茄现有的所有广泛的遗传和基因组资源，转化技术通常被用于促进这些研究。我们实验室常规使用的转化方案已应用于许多不同的番茄基因型，该方案依赖于根癌农杆菌对幼嫩子叶切片的感染。我们使用了用于过表达的载体系统、用于基因沉默的RNA干扰以及用于基因组编辑的CRISPR/Cas9。用于设计基因构建体的载体包含赋予对卡那霉素、潮霉素和除草剂草铵膦抗性的选择标记基因。本章详细介绍了我们用于番茄栽培种和野生种的农杆菌介导转化的方案。
