Illumination of the transmission mechanism of Soil-borne wheat mosaic virus by its vector, Polymyxa graminis and identification of novel resistance factors in barley

小麦多粘菌载体阐明土传小麦花叶病毒的传播机制及大麦中新型抗性因子的鉴定

• 批准号: 403764713
• 负责人: Dr. Annette Niehl
• 金额: --
• 依托单位: Institut für Epidemiologie und Pathogendiagnostik
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2018
• 资助国家: 德国
• 起止时间: 2017-12-31 至 2021-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/403764713?language=en
• 关键词: Illumination; transmission; mechanism; Soil; borne

Polymyxa graminis-transmitted soil-borne viruses cause severe losses in cereals worldwide. As the viruses remain infectious inside polymyxa resting spores for decades, cereal cultivation on contaminated soil is only economic by growing resistant cultivars. Unfortunately, resistances to these viruses are readily overcome by the viruses, thus necessitating innovative resistance strategies. One way to identify novel resistance strategies is the illumination of the molecular mechanisms underlying the interaction between polymyxa-transmitted soil-borne viruses, thereby identifying candidates for virus control by the targeted modulation of host processes involved in infection. Therefore, the objective of the proposed work is the identification of key factors determining the P. graminis - Soil-borne wheat mosaic virus (SBWMV) – barley interaction. As virus transmission and movement are key processes influencing infection, the work specifically elucidates the function and interaction network of the SBWMV transmission factor CP-RT and the SBWMV movement protein (MP). The previewed work includes a detailed analysis of the virus transmission process and virus movement between root cells. In particular, we will investigate interaction between P. graminis and barley root cells by high-resolution in vivo fluorescence microscopy with specific attention on membrane and cytoskeletal rearrangements occurring during the infection process and during virus transmission. Moreover, we will uncover the intra- and intercellular transport pathways the viral MP and CP-RT proteins use to transport viral infectious RNA across the plasmodium membrane and between root cells. Finally, we will use life cell microscopy and immunolocalization in combination with methods for virus quantification to investigate whether virions or viral components are present inside the vector and whether the virus is able to replicate inside polymyxa zoospores. In parallel to these cell biological approaches, we will identify and functionally characterize MP- and CP-RT-interacting host factors in regard to a role in antiviral resistance. Candidate host factors interacting with SBWMV MP and CP-RT proteins will be identified by co-immunoprecipitation followed by mass spectroscopy and a role of the candidate host factors in virus infection will be determined by quantification of vector and virus levels in plants with reduced candidate gene expression. The proposed work will significantly advance our understanding of the molecular mechanisms underlying the interaction between P. graminis, SBWMV and barley in particular and the transmission and infection mechanisms of polymyxa-transmitted soil borne viruses in general. By identifying novel resistance factors in the natural SBWMV host barley, the project will directly contribute to the development of sustainable solutions for agriculture by allowing direct inclusion of the identified resistance factors into resistance breeding programs.

禾谷多粘虫（Polymyxa graminis）通过土传病毒传播，在全球范围内造成严重的谷物损失。由于病毒在多粘菌休眠孢子内保持感染性数十年，在受污染的土壤上种植谷物只有通过种植抗性品种才是经济的。不幸的是，对这些病毒的抗性很容易被病毒克服，因此需要创新的抗性策略。确定新的抗性策略的一种方法是阐明多粘菌传播的土传病毒之间相互作用的分子机制，从而通过有针对性地调节感染中涉及的宿主过程来确定病毒控制的候选者。因此，本研究的目的是确定决定禾谷缢管蚜-土传小麦花叶病毒（SBWMV）-大麦互作的关键因素。由于病毒的传播和运动是影响感染的关键过程，本工作具体阐明了SBWMV传播因子CP-RT和SBWMV运动蛋白（MP）的功能和相互作用网络。预览的工作包括病毒传播过程和根细胞之间的病毒运动的详细分析。特别是，我们将调查P. graminis和大麦根细胞之间的相互作用，通过高分辨率在体内荧光显微镜，特别注意在感染过程中和病毒传播过程中发生的膜和细胞骨架重排。此外，我们将揭示病毒MP和CP-RT蛋白用于跨原质团膜和根细胞之间运输病毒感染性RNA的细胞内和细胞间运输途径。最后，我们将使用生命细胞显微镜和免疫定位与病毒定量方法相结合，以调查是否病毒粒子或病毒成分存在于载体内，以及病毒是否能够在多粘菌游动孢子内复制。在平行于这些细胞生物学方法，我们将确定和功能特点MP和CP-RT相互作用的主机因素方面的作用，抗病毒耐药。与SBWMV MP和CP-RT蛋白相互作用的候选宿主因子将通过免疫共沉淀随后质谱法鉴定，并且候选宿主因子在病毒感染中的作用将通过定量具有降低的候选基因表达的植物中的载体和病毒水平来确定。本研究将有助于我们进一步了解禾谷缢管蚜、小麦花叶病毒和大麦之间相互作用的分子机制，以及多粘菌传播的土传病毒的传播和侵染机制。通过鉴定天然SBWMV宿主大麦中的新抗性因子，该项目将通过允许将鉴定的抗性因子直接纳入抗性育种计划，直接有助于农业可持续解决方案的开发。