School of Life Sciences, Zhejiang Chinese Medical University, Hangzhou 310051, China.
Future Health Laboratory, Innovation Center of Yangtze River Delta, Zhejiang University, Jiaxing 314102, China.
Int J Mol Sci. 2024 Sep 24;25(19):10286. doi: 10.3390/ijms251910286.
is a widespread soil-borne fungal pathogen that can infect various plants, causing wilt and root rot diseases. The root rot disease of caused by is among the most serious diseases associated with continuous cropping, significantly hindering its sustainable development. In this study, we aimed to investigate the effect of exogenous application of double-stranded RNA (dsRNA) on silencing the gene to reduce its virulence and to evaluate its potential application in controlling root rot disease in . The gene was amplified from the genome, and different lengths of -dsRNA were designed and synthesized. The uptake of dsRNA by the fungus was verified using -dsRNA labeled with fluorescein, and in vitro dsRNA treatment experiments were conducted to assess its impact on the growth and virulence of . Additionally, -dsRNA was applied to the roots of to evaluate its effectiveness in controlling root rot disease. The experimental results showed that could effectively uptake exogenously applied -dsRNA, significantly reducing gene expression. All lengths of -dsRNA inhibited fungal growth and caused morphological changes in the fungal hyphae. Further plant experiments and Reverse Transcription Quantitative Polymerase Chain Reaction (RT-qPCR) analysis indicated that -dsRNA treatment significantly reduced the incidence of root rot disease in , which was supported by the reduction in peroxidase (POD) and catalase (CAT) enzyme activities, malondialdehyde (MDA) content, and proline (Pro) levels in treated root tissues. This study demonstrated that exogenous dsRNA could reduce the virulence of by silencing the gene and effectively mitigate the root rot disease it causes in . The successful application of -dsRNA provided strong evidence for the potential of RNA interference (RNAi) technology in plant disease control. Future research could further optimize the design and application of dsRNA to enhance its practical value in agriculture.
是一种广泛存在的土壤传播真菌病原体,可感染各种植物,引起萎蔫和根腐病。由 引起的根腐病是与连作相关的最严重疾病之一,严重阻碍了其可持续发展。在本研究中,我们旨在研究外源双链 RNA(dsRNA)对沉默 基因以降低其毒力的影响,并评估其在控制 根腐病中的潜在应用。从 基因组中扩增了 基因,并设计和合成了不同长度的 -dsRNA。通过用荧光素标记的 -dsRNA 验证了真菌对 dsRNA 的摄取,并进行了体外 dsRNA 处理实验,以评估其对 的生长和毒力的影响。此外,将 -dsRNA 应用于 的根部,以评估其控制根腐病的效果。实验结果表明, 可以有效地摄取外源施加的 -dsRNA,显著降低 基因表达。所有长度的 -dsRNA 均抑制了真菌的生长,并导致真菌菌丝的形态发生变化。进一步的植物实验和反转录定量聚合酶链反应(RT-qPCR)分析表明,-dsRNA 处理显著降低了 的根腐病发病率,这得到了处理根组织中过氧化物酶(POD)和过氧化氢酶(CAT)酶活性、丙二醛(MDA)含量和脯氨酸(Pro)水平降低的支持。本研究表明,外源 dsRNA 通过沉默 基因可以降低 的毒力,并有效地减轻其引起的根腐病。-dsRNA 的成功应用为 RNA 干扰(RNAi)技术在植物病害控制中的应用提供了有力证据。未来的研究可以进一步优化 dsRNA 的设计和应用,以提高其在农业中的实际价值。