Zhang Ting, Hou Jiaqi, Chu Hongtao, Guo Pengyu, Sang Qianzi, Liu Zhongxu, Cao Li
Agricultural College, Yanbian University, Yanji 133002, China.
Laboratory of Molecular Biology of Tomato, Bioengineering College, Chongqing University, Chongqing 400044, China.
Plants (Basel). 2025 Jan 7;14(2):150. doi: 10.3390/plants14020150.
L. (Jinhuakui, JHK) is widely cultivated for its pharmacological properties owing to its high flavonoid content and is commonly used as a garden landscape plant. However, the absence of an efficient genetic transformation system poses significant challenges for functional gene studies in this species. Virus-induced gene silencing (VIGS) is a well-established technique for exploring plant gene functions; however, this technique has not been applied to JHK. Here, a tobacco rattle virus (TRV)-VIGS system was successfully developed for the first time in JHK using the gene encoding phytoene desaturase () as a marker gene. This study investigated the impact of various infection methods on the efficiency of silencing. The results demonstrated that administering two injections-the first on the day of complete cotyledon expansion and the second 14 days later-using pTRV1 and pTRV2- cultures resuspended to an OD of 1.0 and via the backside of the blade-led to significant photobleaching in the cotyledons 2 days after the second injection. Subsequent analyses revealed a marked reduction in both chlorophyll content and expression. These findings suggest that a VIGS system was successfully developed in JHK, thus providing a rapid and effective method for studying gene function in this species and facilitating future research in JHK genetics.
金花葵（Jinhuakui，JHK）因其高黄酮含量的药理特性而被广泛种植，并且通常被用作园林景观植物。然而，缺乏高效的遗传转化系统给该物种的功能基因研究带来了重大挑战。病毒诱导的基因沉默（VIGS）是一种成熟的用于探索植物基因功能的技术；然而，该技术尚未应用于金花葵。在此，首次在金花葵中成功开发了一种烟草脆裂病毒（TRV）-VIGS系统，使用编码八氢番茄红素去饱和酶（）的基因作为标记基因。本研究调查了各种感染方法对沉默效率的影响。结果表明，使用重悬至OD值为1.0的pTRV1和pTRV2培养物，在子叶完全展开当天进行第一次注射，14天后进行第二次注射，并通过叶片背面注射，在第二次注射后2天导致子叶出现明显的光漂白现象。随后的分析显示叶绿素含量和表达均显著降低。这些发现表明在金花葵中成功开发了VIGS系统，从而为研究该物种的基因功能提供了一种快速有效的方法，并促进了金花葵遗传学的未来研究。
