Plant vacuoles are dynamic organelles that play essential roles in regulating growth and development. Two distinct models of vacuole biogenesis have been proposed: separate vacuoles are formed by the fusion of endosomes, or the single interconnected vacuole is derived from the endoplasmic reticulum. These two models are based on studies of two-dimensional (2D) transmission electron microscopy and 3D confocal imaging, respectively. Here, we performed 3D electron tomography at nanometre resolution to illustrate vacuole biogenesis in Arabidopsis root cells. The whole-cell electron tomography analysis first identified unique small vacuoles (SVs; 400–1,000 nm in diameter) as nascent vacuoles in early developmental cortical cells. These SVs contained intraluminal vesicles and were mainly derived/matured from multivesicular body (MVB) fusion. The whole-cell vacuole models and statistical analysis on wild-type root cells of different vacuole developmental stages demonstrated that central vacuoles were derived from MVB-to-SV transition and subsequent fusions of SVs. Further electron tomography analysis on mutants defective in MVB formation/maturation or vacuole fusion demonstrated that central vacuole formation required functional MVBs and membrane fusion machineries.
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This work was supported by grants from the Research Grants Council of Hong Kong (CUHK14130716, 14102417, 14100818, C4011-14R, C4012-16E, C4002-17G and AoE/M-05/12) and the National Natural Science Foundation of China (31270226, 31470294 and 91854201).
The authors declare no competing interests.
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Supplementary Figures 1–8.
Whole-cell electron tomography analyses of SVs in relationship with other organelles in Cell 1.
3D tomography analyses of detailed structures and relationships of ER, Golgi, TGN, MVBs and SVs.
3D tomography analyses of fusion between MVBs and SVs, along with a transfer of ILVs.
Whole-cell electron tomography analyses of vacuoles in Cell 2.
Whole-cell electron tomography analyses of vacuoles in Cell 3.
3D tomography analyses of vacuoles in WT and various mutants.
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Cui, Y., Cao, W., He, Y. et al. A whole-cell electron tomography model of vacuole biogenesis in Arabidopsis root cells. Nature Plants 5, 95–105 (2019). https://doi.org/10.1038/s41477-018-0328-1
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