Genetic transformation has always been an important method for studying medical plant secondary metabolic regulation, among which stable transformation has a good reproducibility. However, it was time-consuming to obtain a stable transformed hairy root or transgenic plants, which was difficult to satisfy the great demand of researches on medical plant secondary metabolism–related genes. Moreover, Agrobacterium tumefaciens–mediated transient transformation has been extensively applied in studies of functional genes because of its simpleness, low cost, and short period. However, presently, researches on medical plant functional genes commonly used stable genetic transformation and some high-cost and high-difficulty transient transformation methods, such as gene gun and protoplast transformation. Thus, in this study, we selected the seedlings of Nicotiana benthamiana, Salvia miltiorrhiza, and Prunella vulgaris as the experimental material, with the methods of Agrobacterium tumefaciens injection, fast Agrobacterium-mediated seedling transformation (FAST), and FAST and mechanical damage. The results demonstrated that the injection transient transformation system of pCAMBIA1301 vector mediated by A. tumefaciens and the transient transformation of seedling system were not established in S. miltiorrhiza. In addition, the instantaneous transformation system of N. benthamiana and P. vulgaris seedlings was basically set up by FAST method. Besides, using the method of FAST and mechanical damage, the transient genetic transformation system of P. vulgaris seedlings was established for the first time. A. tumefaciens–mediated transient transformation of seedlings with pEAQ vectors provided an effective way and reference for the further study of functional genes of the medicinal plants N. benthamiana and P. vulgaris.

中文翻译：

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建立农杆菌介导的药用植物瞬时表达系统的尝试

遗传转化一直是研究药用植物次生代谢调控的重要方法，其中稳定转化具有良好的重现性。然而，获得稳定的转化毛状根或转基因植物非常耗时，难以满足药用植物次生代谢相关基因研究的巨大需求。此外，根癌农杆菌介导的瞬时转化因其简单、成本低、周期短等优点而被广泛应用于功能基因的研究。但目前药用植物功能基因的研究多采用稳定遗传转化和一些高成本、高难度的瞬时转化方法，如基因枪、原生质体转化等。因此，在本研究中，选取本氏烟草、丹参、夏枯草幼苗为实验材料，采用根癌农杆菌注射法、农杆菌介导的快速幼苗转化法（FAST）和机械损伤法。结果表明，根癌农杆菌介导的pCAMBIA1301载体注射瞬时转化系统和苗木瞬时转化系统均未在丹参中建立。此外，本氏烟草和普通白杨幼苗的瞬时转化体系基本采用FAST方法建立。此外，利用FAST和机械损伤的方法，首次建立了P. vulgaris幼苗瞬时遗传转化系统。一种。