El-Tanbouly Rania, Hassan Hend, Awd Lojina M, Makhlouf Azza Ali, Shalabi Hani G, El-Messeiry Sarah
Department of Floriculture, Ornamental Horticulture and Landscape Design, Faculty of Agriculture, Alexandria University, 163 Aflaton Street, El Shatby, Alexandria, 21545, Egypt.
Department of Genetics, Faculty of Agriculture, Alexandria University, 163 Aflaton Street, El Shatby, Alexandria, 21545, Egypt.
Heliyon. 2024 Nov 21;10(23):e40589. doi: 10.1016/j.heliyon.2024.e40589. eCollection 2024 Dec 15.
Medical plants play a crucial role in the pharmaceutical industry due to their natural synthesis of active compounds. Synthetic methods exist, which provide fewer effective molecules compared to those naturally occurring. , a significant medicinal plant, synthesizes vital vinca alkaloids and various secondary metabolites widely used in cancer and hypertension treatment. However, genetic modification using -mediated transformation, a common method for altering periwinkle plants, suffers from low efficiency and reproducibility. Factors like the Agrobacterium strain can influence transformation efficiency and post-transformation regeneration. This study compares the transformation and regeneration efficiency of three A. tumefaciens strains (LBA4404, EHA105, and GV3101) carrying the GUS gene and kanamycin selection via syringe infiltration in . Molecular variations between mutants were examined using Inter-Simple Sequence Repeat (ISSR) and GUS expression via quantitative Real-Time PCR (qRT-PCR). Results revealed that GV3101 had the highest transformation efficiency (61.1 %) and LBA4404 the lowest (38 %). However, GV3101-infected explants had the lowest regeneration rate (10 %), the obtained mutants from it exhibited the highest GUS expression. ISSR analysis indicated 37 % polymorphism among mutants, highlighting the impact of Agrobacterium strains on plant genetics and potentially on phytochemical compositions. Overall, this study recommends using GV3101 for high transformation efficiency and LBA4404 for superior regeneration in , suggesting a promising method for efficient gene transformation in this plant species.
药用植物因其能天然合成活性化合物而在制药行业中发挥着关键作用。虽然存在合成方法,但与天然存在的分子相比,合成方法产生的有效分子较少。长春花是一种重要的药用植物,能合成重要的长春花生物碱和各种次生代谢产物,广泛用于癌症和高血压治疗。然而,利用农杆菌介导的转化进行基因改造是改变长春花植物的常用方法,但该方法效率低且重复性差。诸如农杆菌菌株等因素会影响转化效率和转化后的再生。本研究比较了携带GUS基因并通过注射器浸润法在长春花中进行卡那霉素筛选的三种根癌农杆菌菌株(LBA4404、EHA105和GV3101)的转化和再生效率。使用简单序列重复区间(ISSR)检测突变体之间的分子变异,并通过定量实时聚合酶链反应(qRT-PCR)检测GUS表达。结果显示,GV3101的转化效率最高(61.1%),LBA4404最低(38%)。然而,感染GV3101的外植体再生率最低(10%),从中获得的突变体GUS表达最高。ISSR分析表明突变体间存在37%的多态性,突出了农杆菌菌株对植物遗传学以及可能对植物化学成分的影响。总体而言,本研究建议在长春花中使用GV3101以获得高转化效率,使用LBA4404以获得更好的再生效果,这为该植物物种的高效基因转化提供了一种有前景的方法。