J Integr Plant Biol. ›› 2021, Vol. 63 ›› Issue (2): 353-364.DOI: 10.1111/jipb.13027

Special Issue: Biotechnology Plant-biotic interaction

• Research Articles • Previous Articles     Next Articles

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

Xueting Wang1, Juncai Ma2, Xuejiao Jin1, Ning Yue1, Peng Gao2, Keith Ka Ki Mai2, Xian‐Bing Wang1, Dawei Li1, Byung‐Ho Kang2* and Yongliang Zhang1*   

  1. 1State Key Laboratory of Agro‐Biotechnology and Ministry of Agriculture Key Laboratory of Soil Microbiology, College of Biological Sciences, China Agricultural University, Beijing 100193, China
    2State Key Laboratory of Agro‐Biotechnology, School of Life Sciences, The Chinese University of Hong Kong, Hong Kong 999077, China

    *Correspondences: Byung-Ho Kang (bkang@cuhk.edu.hk); Yongliang Zhang(cauzhangyl@cau.edu.cn, Dr. Zhang is fully responsible for the distribution of all materials associated with this article)
  • Received:2020-09-15 Accepted:2020-10-20 Online:2020-10-21 Published:2021-02-01

Abstract: 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 1a and 2a 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.

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