Faion-Molina Mayra, Molina-Risco Marco David, Bellinatti-Della Gracia Marcela Domingues, Ibarra Oneida, Kim Backki, Septiningsih Endang M, Thomson Michael J
Department of Soil and Crop Sciences, Texas A&M University, College Station, TX, USA.
Avance Biosciences Inc., Houston, TX, USA.
Methods Mol Biol. 2025;2911:45-59. doi: 10.1007/978-1-0716-4450-8_6.
Bottlenecks in plant transformation and regeneration have slowed progress in applying CRISPR/Cas9-based genome editing for crop improvement. Rice (Oryza sativa L.) has highly efficient temperate japonica transformation protocols, along with reasonably efficient indica protocols using immature embryos. However, rapid and efficient protocols are not available for transformation and regeneration in tropical japonica varieties, even though they represent most of the rice production in the USA and South America, along with some regions in Asia. This chapter describes a protocol for CRISPR/Cas9 gene editing using Agrobacterium-mediated transformation for the tropical japonica rice cultivar Presidio leading to knock-out mutations in the phytoene desaturase (PDS) gene.
植物转化和再生过程中的瓶颈减缓了基于CRISPR/Cas9的基因组编辑技术在作物改良中的应用进展。水稻（Oryza sativa L.）拥有高效的温带粳稻转化方案，以及使用未成熟胚的较为高效的籼稻方案。然而，热带粳稻品种的转化和再生尚无快速高效的方案，尽管它们在美国、南美洲以及亚洲的一些地区占据了大部分水稻产量。本章描述了一种利用农杆菌介导的转化对热带粳稻品种Presidio进行CRISPR/Cas9基因编辑的方案，该方案可导致八氢番茄红素去饱和酶（PDS）基因产生敲除突变。
