Abstract:
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.
摘要:
对体外培养和再生的依赖性限制了对优质小麦(TriticumaestivumL.)品种使用基因组编辑的能力。我们最近开发了一种用于小麦基因编辑的植物粒子轰击(iPB)技术,该技术利用茎尖分生组织(SAMs)作为靶组织。由于该方法不需要体外培养,因此,它可以用于顽固的品种。在这一章中,我们详细描述了iPB方法中使用的步骤。有了这个协议,从轰击的SAM生长的T0植物的3%至5%通常携带突变等位基因,并且大约1%至2%的T0植物在下一代中继承突变等位基因。