Agrobacterium tumefaciens-Mediated Transformation of Tomato.
Agrobacterium tumefaciens-Mediated Transformation of Tomato.
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DOI：
10.1007/978-1-4939-8778-8_16
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发表时间：
2018-11
影响因子：
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通讯作者：
J. Van Eck;Patricia Keen;Michelle Tjahjadi
中科院分区：
文献类型：
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作者：
J. Van Eck;Patricia Keen;Michelle Tjahjadi
番茄既是一种重要的粮食作物，也是一种模式植物，用于各种研究调查，包括了解基因功能。转化通常用于促进这些调查与所有广泛的遗传和基因组资源的番茄。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，并且依赖于嫩子叶切片的根癌农杆菌感染。我们已经使用载体系统进行过表达，RNA干扰用于基因沉默，CRISPR/Cas9用于基因组编辑。用于设计基因构建体的载体含有可选择标记基因，其赋予对卡那霉素、潮霉素和除草剂组分双丙氨膦的抗性。本章详细介绍了农杆菌介导的番茄栽培种和野生种的转化方法。
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 onAgrobacterium tumefaciensinfection 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 forAgrobacterium-mediated transformation of both cultivated and wild species of tomato is detailed in this chapter.