Multifactorial analysis of simultaneous organelle movement reveals cell-specific motility of peroxisomes and mitochondria.

The movement, distribution, and interactions of organelles are cell-type specific, responding to fluctuating metabolic and environmental cues and governing the efficiency of plant physiology and stress response. The directional motility of various plant organelles is predominantly driven by the actomyosin system, yet the distinct functionality of these organelles across plant tissues presupposes organelle-specific regulation of motility, which requires the detection of subtle shifts in dynamics. Meanwhile, studies that comprehensively characterize and directly compare the simultaneous movement of multiple types of organelles within the same cell are limited. Here, we visualized peroxisomes, mitochondria, chloroplasts, Golgi bodies, and actin filaments simultaneously in tobacco (Nicotiana tabacum) to evaluate organelle organization and motility within the context of one another. Quantitative analysis of multiple motility factors enabled us to identify peroxisome motility in tobacco mesophyll as distinct from other organelles. Further analysis in Arabidopsis (Arabidopsis thaliana) revealed that both mitochondria and peroxisomes are slower in mesophyll cells compared to epidermis in normal growth conditions, but their motility patterns are unique from one another across leaf tissue after plants experienced conditions that induce photorespiration, a metabolic pathway requiring the concerted action of chloroplasts, peroxisomes, and mitochondria. Our quantitative analysis of thousands of organelles across species, cell type, and physiological conditions unveils distinct modulation of motility according to organelle identity and function. The extensive combinatorial characterizations of plant organelle movement provide a fundamental resource for the future discovery of molecular mechanisms driving the movement and distribution of diverse organelles.