Division of Crop Genome Editing, Institute of Agrobiological Sciences, National Agriculture and Food Research Organization (NARO), Tsukuba, Japan.
Institute of Plant and Food Science, Department of Biology, Southern University of Science and Technology, Shenzhen, China.
Methods Mol Biol. 2024;2830:163-171. doi: 10.1007/978-1-0716-3965-8_15.
Dependency on in vitro culture and regeneration limits the ability to use genome editing on elite wheat (Triticum aestivum L.) varieties. We recently developed an in planta particle bombardment (iPB) technique for gene editing in wheat that utilizes shoot apical meristems (SAMs) as a target tissue. Since the method does not require in vitro culture, it can therefore be used on recalcitrant varieties. In this chapter, we describe in detail the steps used in the iPB method. With this protocol, 3% to 5% of T0 plants grown from bombarded SAMs typically carry mutant alleles and approximately 1% to 2% of the T0 plants inherit mutant alleles in the next generation.
对体外培养和再生的依赖限制了利用基因组编辑技术对优质小麦（Triticum aestivum L.）品种进行改良的能力。我们最近开发了一种在小麦中进行基因编辑的体内粒子轰击（iPB）技术，该技术利用茎尖分生组织（SAM）作为靶组织。由于该方法不需要体外培养，因此可以用于顽固品种。在本章中，我们详细描述了 iPB 方法中使用的步骤。使用该方案，从轰击的 SAM 中生长的 T0 植物中约有 3%至 5%通常携带突变等位基因，而约 1%至 2%的 T0 植物在下一世代中继承突变等位基因。
