SummarySucculents, valued for their drought tolerance and ornamental appeal, are important in the floriculture market. However, only a handful of succulent species can be genetically transformed, making it difficult to improve these plants through genetic engineering. In this study, we adapted the recently developed cut‐dip‐budding (CDB) gene delivery system to transform three previously recalcitrant succulent varieties – the dicotyledonous Kalanchoe blossfeldiana and Crassula arborescens and the monocotyledonous Sansevieria trifasciata. Capitalizing on the robust ability of cut leaves to regenerate shoots, these plants were successfully transformed by directly infecting cut leaf segments with the Agrobacterium rhizogenes strain K599. The transformation efficiencies were approximately 74%, 5% and 3.9%–7.8%, respectively, for K. blossfeldiana and C. arborescens and S. trifasciata. Using this modified CDB method to deliver the CRISPR/Cas9 construct, gene editing efficiency in K. blossfeldiana at the PDS locus was approximately 70%. Our findings suggest that succulents with shoot regeneration ability from cut leaves can be genetically transformed using the CDB method, thus opening up an avenue for genetic engineering of these plants.

中文翻译：

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一种无需组织培养的多肉植物遗传转化和基因编辑方法

摘要 多肉植物因其耐旱性和观赏性而受到重视，在花卉市场上很重要。然而，只有少数多肉植物品种可以进行基因改造，因此很难通过基因工程来改良这些植物。在这项研究中，我们采用了最近开发的切浸芽接（CDB）基因传递系统来改造三个以前顽固的多肉品种——双子叶植物长寿花和木立景天和单子叶植物三带虎尾兰。利用切叶再生芽的强大能力，这些植物通过直接感染切叶段而成功转化发根农杆菌菌株K599。转化效率分别约为 74%、5% 和 3.9%–7.8%布氏堇菜和木立竹和三带龙舌兰。使用这种改进的 CDB 方法来传递 CRISPR/Cas9 构建体，可提高基因编辑效率布氏堇菜在PDS位点约为70%。我们的研究结果表明，具有切叶再生能力的多肉植物可以使用CDB方法进行遗传转化，从而为这些植物的基因工程开辟了一条途径。