Phytophthora capsici, which causes diseases in solanaceous crops, secretes necrosis and ethylene-inducing peptide 1-like proteins (NLPs) that induce plant defense responses and leaf necrosis. In this study, we used RNA interference (RNAi) technique, a proven strategy for crop protection and gene regulation in plants, to suppress P. capsici infection through the inhibition of PcNLPs. In the RNAi mechanism, Dicer processes double-stranded RNA (dsRNA) into smaller entities known as small interfering RNAs (siRNAs). These siRNAs subsequently integrate into the RNA-induced silencing complex to form sequence-specific base pairing with complementary regions of the target mRNA. This interaction effectively initiates the degradation process of the target mRNA. We designed and synthesized dsRNAs targeting the “AIMY” and “GHRHDWE” conserved motifs of PcNLP gene family, which are predicted to be key elements for the expression of NLPs and pathogen infection. After infiltration of dsRNAs targeting the motifs and inoculation with P. capsici, we confirmed a significant suppression of P. capsici infection and downregulation of the PcNLP gene family. These findings imply that the dsRNA-mediated RNAi technique holds potential for mitigating a wide range of pathogens, while simultaneously suppressing the expression of a particular gene family using dsRNA targeting functional conserved motifs in the gene family.

中文翻译：

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开发针对 PcNLP 基因家族的多靶点 dsRNA 以抑制烟草疫霉感染

辣椒疫霉会引起茄科作物的疾病，它会分泌坏死蛋白和乙烯诱导肽 1 样蛋白 (NLP)，从而诱导植物防御反应和叶片坏死。在这项研究中，我们使用 RNA 干扰 (RNAi) 技术，这是一种经过验证的作物保护和植物基因调控策略，通过抑制 PcNLP 来抑制辣椒疫病菌感染。在 RNAi 机制中，Dicer 将双链 RNA (dsRNA) 加工成更小的实体，称为小干扰 RNA (siRNA)。这些 siRNA 随后整合到 RNA 诱导的沉默复合物中，与靶 mRNA 的互补区域形成序列特异性碱基配对。这种相互作用有效地启动了目标 mRNA 的降解过程。我们设计并合成了针对 PcNLP 基因家族“AIMY”和“GHRHDWE”保守基序的 dsRNA，预计这些基序将成为 NLP 表达和病原体感染的关键元件。在针对该基序的 dsRNA 渗透并接种辣椒疫霉后，我们证实了辣椒疫霉感染的显着抑制和 PcNLP 基因家族的下调。这些发现表明，dsRNA 介导的 RNAi 技术具有减轻多种病原体的潜力，同时使用 dsRNA 靶向基因家族中的功能性保守基序来抑制特定基因家族的表达。