ABSTRACT In plant biology, transient expression analysis plays a vital role to provide a fast method to study the gene of interest. In this study, we report a rapid and efficient method for transient expression in Cannabis sativa seedlings using Agrobacterium tumefaciens-mediated transformation. A. tumefaciens strain EHA105 carrying the pCAMBIA1301 construct with uidA gene was used to transform cannabis seedlings and the GUS assay (a measurement of β-glucuronidase activity) was used to detect the uidA expression. In the current study, we have also established a rapid germination protocol for cannabis seeds. The all three steps seed sterilization, germination and seedlings development were carried out in a 1% H2O2 solution. Transient transformation revealed that both cotyledons and young true leaves are amenable to transformation. Compared with tobacco (Nicotiana benthamiana), cannabis seedlings were less susceptible to transformation with A. tumefaciens. Susceptibility to Agrobacterium transformation also varied with the different cannabis varieties. The method established in this study has the potential to be an important tool for gene function studies and genetic improvement in cannabis.

中文翻译：

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β-葡萄糖醛酸酶基因在大麻品种中的瞬时表达

摘要 在植物生物学中，瞬时表达分析在提供一种快速方法来研究感兴趣的基因方面起着至关重要的作用。在这项研究中，我们报告了一种使用根癌农杆菌介导的转化在大麻幼苗中进行瞬时表达的快速有效方法。携带带有 uidA 基因的 pCAMBIA1301 构建体的根癌农杆菌菌株 EHA105 用于转化大麻幼苗，并使用 GUS 测定（β-葡萄糖醛酸酶活性的测量）检测 uidA 表达。在目前的研究中，我们还建立了大麻种子的快速发芽方案。种子灭菌、发芽和幼苗发育这三个步骤均在 1% H2O2 溶液中进行。瞬时转化表明子叶和幼叶都适合转化。与烟草（本氏烟草）相比，大麻幼苗不易被根癌农杆菌转化。对农杆菌转化的敏感性也因不同的大麻品种而异。本研究中建立的方法有可能成为大麻基因功能研究和遗传改良的重要工具。