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Three-dimensional reconstruction and comparison of vacuolar membranes in response to viral infection

查看全文 作  者:Xueting [1]Wang;Juncai [2]Ma;Xuejiao [1]Jin;Ning [1]Yue;Peng [2]Gao;Keith Ka Ki [2]Mai;Xian-Bing [1]Wang;Dawei [1]Li;Byung-Ho [2]Kang;Yongliang [1]Zhang 高影响力作者 机构地区:[1]State Key Laboratory of Agro-Biotechnology and Ministry of Agriculture Key Laboratory of Soil Microbiology,College of Biological Sciences,China Agricultural University,Beijing 100193,China;[2]State Key Laboratory of Agro-Biotechnology,School of Life Sciences,The Chinese University of Hong Kong,Hong Kong 999077,China高影响力机构 出  处:《Journal of Integrative Plant Biology》索引2021年第63卷第2期,共12页高影响力期刊 基  金:supported by grants from the National Transgenic Science and Technology Program (2019ZX08010-003);the National Natural Science Foundation of China (31872637);Chinese Universities Scientific Fund (2020TC181);the Research Grants Council of Hong Kong (GRF14126116, GRF14121019, C4012-16E, C4002-17G, and Ao E/M-05/12);Cooperative Research Program for Agriculture Science & Technology Development (0109532019) Rural Development Administration, Republic of Korea。 摘  要:The vacuole is a unique plant organelle that plays an important role in maintaining cellular homeostasis under various environmental stress conditions. However, the effects of biotic stress on vacuole structure has not been examined using three-dimensional(3D) visualization. Here, we performed 3D electron tomography to compare the ultrastructural changes in the vacuole during infection with different viruses. The 3D models revealed that vacuoles are remodeled in cells infected with cucumber mosaic virus(CMV) or tobacco necrosis virus A Chinese isolate(TNV-AC), resulting in the formation of spherules at the periphery of the vacuole. These spherules contain neck-like channels that connect their interior with the cytosol. Confocal microscopy of CMV replication proteins 1 a and 2 a and TNV-AC auxiliary replication protein p23 showed that all of these proteins localize to the tonoplast.Electron microscopy revealed that the expression of these replication proteins alone is sufficient to induce spherule formation on the tonoplast, suggesting that these proteins play prominent roles in inducing vacuolar membrane remodeling. This is the first report of the 3D structures of viral replication factories built on the tonoplasts. These findings contribute to our understanding of vacuole biogenesis under normal conditions and during assembly of plant(+) RNA virus replication complexes. 关 键 词:COMPARISON cucumber mosaic virus electron tomography SPHERULES tobacco necrosis virus A VACUOLE viral replication
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