Department of Plant Pathology, Tamil Nadu Agricultural University, Coimbatore, 641003, India.
Centre for Plant Molecular Biology and Biotechnology, Tamil Nadu Agricultural University, Coimbatore, 641003, India.
Planta. 2023 Oct 7;258(5):94. doi: 10.1007/s00425-023-04253-6.
Exogenous application of dsRNA molecules targeting MYMV genes offers a promising approach to effectively mitigate yellow mosaic disease in blackgram, demonstrating potential for sustainable plant viral disease management. The exogenous application of double-stranded RNA (dsRNA) molecules to control plant viral diseases is gaining traction due to its advantages over conventional methods, such as target specificity, non-polluting nature, and absence of residue formation. Furthermore, this approach does not involve genome modification. In this study, dsRNA molecules targeting the coat protein gene (dsCP) and replication initiator protein gene (dsRep) of mungbean yellow mosaic virus (MYMV) were synthesised using an in vitro transcription method. To evaluate the effectiveness of dsRNA treatment, blackgram plants exhibiting MYMV symptoms at the first trifoliate stage were subjected to exogenous application of dsRNA. Second, third, and fourth trifoliate leaves, which emerged at 7, 15, and 21 days after dsRNA application, respectively, were monitored for MYMV symptoms. Remarkably, a significant reduction in yellow mosaic disease (YMD) symptoms was observed in the newly emerged trifoliate leaves of MYMV-infected blackgram plants after treatment with dsRNA targeting both gene regions. This reduction was evident as a decrease in the intensity of yellow mosaic coverage on the leaf lamina compared to control. dsCP effectively reduced the MYMV titre in the treated plants for up to 15 days. However, dsRep demonstrated greater efficiency in conferring resistance to MYMV at 15 days post-application. These findings were supported by quantitative real-time PCR analysis, where the observed Ct values for DNA extracted from dsRep-treated plants were significantly higher compared to the Ct values of DNA from dsCP-treated plants at 15 days post-application. Similarly, higher viral copy numbers were observed in dsCP-treated plants 15 days after dsRNA treatment, in contrast to plants treated with dsRep.
针对 MYMV 基因的 dsRNA 分子的外源应用为有效减轻菜豆黄花叶病提供了一种有前途的方法，展示了可持续植物病毒病管理的潜力。由于其具有针对目标的特异性、非污染性和无残留形成等优点，外源应用双链 RNA（dsRNA）分子来控制植物病毒病正受到越来越多的关注。此外，这种方法不涉及基因组修饰。在这项研究中，使用体外转录方法合成了针对豇豆花叶病毒（MYMV）外壳蛋白基因（dsCP）和复制起始蛋白基因（dsRep）的 dsRNA 分子。为了评估 dsRNA 处理的有效性，在第一复叶期出现 MYMV 症状的菜豆植株上进行了 dsRNA 的外源应用。然后，分别在 dsRNA 处理后第 7、15 和 21 天出现的第二、第三和第四复叶上监测 MYMV 症状。值得注意的是，在用针对两个基因区域的 dsRNA 处理感染 MYMV 的菜豆植物后，新出现的复叶中明显减轻了黄斑驳病（YMD）症状。与对照相比，叶片叶肉上的黄斑驳覆盖率的强度降低了。dsCP 有效地将处理植物中的 MYMV 滴度降低了 15 天。然而，dsRep 在 dsRNA 处理后 15 天对 MYMV 的抗性效率更高。这些发现得到了定量实时 PCR 分析的支持，其中从 dsRep 处理的植物中提取的 DNA 的观察到的 Ct 值在 dsRNA 处理后 15 天明显高于从 dsCP 处理的植物中提取的 DNA 的 Ct 值。同样，在 dsRNA 处理后 15 天，dsCP 处理的植物中观察到更高的病毒拷贝数，而 dsRep 处理的植物则相反。
