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Abstract：
The cloning of resistance-related genes CsROP5/CsROP10 and the analysis of their mechanism of action provide a theoretical basis for the development of molecular breeding of disease-resistant cucumbers. The structure domains of two Rho-related guanosine triphosphatases from plant (ROP) genes were systematically analyzed using the bioinformatics method in cucumber plants, and the genes CsROP5 (Cucsa.322750) and CsROP10 (Cucsa.197080) were cloned. The functions of the two genes were analyzed using reverse-transcription quantitative PCR (RT-qPCR), virus-induced gene silencing (VIGS), transient overexpression, cucumber genetic transformation, and histochemical staining technology. The conserved elements of the CsROP5/CsROP10 proteins include five sequence motifs (G1-G5), a recognition site for serine/threonine kinases, and a hypervariable region (HVR). The knockdown of CsROP10 through VIGS affected the transcript levels of ABA-signaling-pathway-related genes (CsPYL, CsPP2Cs, CsSnRK2s, and CsABI5), ROS-signaling-pathway-related genes (CsRBOHD and CsRBOHF), and defense-related genes (CsPR2 and CsPR3), thereby improving cucumber resistance to Corynespora cassiicola. Meanwhile, inhibiting the expression of CsROP5 regulated the expression levels of ROS-signaling-pathway-related genes (CsRBOHD and CsRBOHF) and defense-related genes (CsPR2 and CsPR3), thereby enhancing the resistance of cucumber to C. cassiicola. Overall, CsROP5 and CsROP10 may participate in cucumber resistance to C. cassiicola through the ROS and ABA signaling pathways.
摘要：
抗病相关基因CsROP5/CsROP10的克隆及其作用机理分析为开展抗病黄瓜的分子育种提供了理论依据。利用生物信息学方法对黄瓜植物中两个与Rho相关的三磷酸鸟苷(ROP)基因的结构域进行了系统分析,克隆了CsROP5(Cucsa.322750)和CsROP10(Cucsa.197080)基因。使用逆转录定量PCR(RT-qPCR)分析这两个基因的功能,病毒诱导的基因沉默（VIGS），瞬时过表达,黄瓜遗传转化,和组织化学染色技术。CsROP5/CsROP10蛋白的保守元件包括五个序列基序(G1-G5),丝氨酸/苏氨酸激酶的识别位点，和高变区（HVR）。通过VIGS敲低CsROP10会影响ABA信号通路相关基因的转录水平（CsPYL，CsPP2C,CsSnRK2s,和CsABI5），ROS信号通路相关基因（CsRBOHD和CsRBOHF），和防御相关基因（CsPR2和CsPR3），从而提高黄瓜对Corynessporacassiicola的抗性。同时,抑制CsROP5的表达调节了ROS信号通路相关基因（CsRBOHD和CsRBOHF）和防御相关基因（CsPR2和CsPR3）的表达水平，从而增强黄瓜对C.cassiicola的抗性。总的来说,CsROP5和CsROP10可能通过ROS和ABA信号通路参与了黄瓜对Cassiicola的抗性。
