重磅突破！利用叶片就可以获得转基因及基因编辑植株的新方法，涵盖玉米、大麦、高粱等9种禾本科植物

近日，Nature plants在线发表了题为“Leaf transformation for efficient random integration and targeted genome modification in maize and sorghum”的研究论文，该研究报道了一种利用叶片既可以获得转基因及基因编辑植株的方法，并且在大麦，玉米，水稻，黑麦草等9种禾本科植物中得到验证。

传统上，禾本科物种的转化依赖于不成熟的胚胎，因此只限于几种主要的禾本科作物。其他的转化的外植体，包括叶片组织，也曾被探索过，但成功率很低，这是阻碍基因编辑在作物遗传改良中的广泛应用的主要因素之一。最近，利用Wuschel2（Wus2）和Babyboom（Bbm）两个基因的的叶片转化已成功用于Cas9介导的突变，但仍然需要大量再生植物筛选。在该研究中，作者证明了增强的Wus2/Bbm表达显著提高了玉米和高粱的叶片转化，并且获得了Cas9介导基因编辑植株。此外，使用针对玉米优化的Wus2/Bbm构建，在涵盖四个禾本科亚科的八个物种中成功地生成了胚性愈伤组织和再生植株。

Fig. 3: The leaf transformation method used for maize.

a, Surface sterilization of maize seeds. b, Maize seeds on germination medium. c, Fourteen-day-old maize seedlings used as the explant source. d, Preparation of 3 cm segments of the leaf whorl directly above the mesocotyl followed by slicing the tissue longitudinally (not shown). e, The tissue was placed in 100 ml of Agrobacterium suspension in the mini-Cuisinart blender and subjected to ten brief pulses to further slice up the leaf tissue. After a 20-minute incubation in the Agrobacterium suspension, the liquid was poured through a metal sieve to collect the leaf fragments, which were briefly blotted onto dry filter paper to wick away excess Agrobacterium (not shown). f, Leaf fragments distributed onto filter paper on the solid RM. g, Fluorescent somatic embryo growth observed 7–10 days after Agrobacterium infection (the image is representative of over 100 independent experiments; scale bar, 1 mm). h, After culturing on the RM, leaf tissue was transferred to selection medium. i, Following three weeks on filter papers on selection medium, the tissue was subjected to 45 °C/70% RH for two hours (not shown), transferred to maturation medium and cultured for an additional three weeks. j, Formation of T0 regenerants after two weeks on rooting medium.

原文链接：

https://www.nature.com/articles/s41477-022-01338-0

https://www.nature.com/articles/s41477-023-01349-5

转自：“植物生物技术Pbj”微信公众号

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