Myosin-Driven Nuclear Movement Generates Stromules in Arabidopsis thaliana

Plastids form stroma-filled tubular extensions, termed stromules, that are implicated in inter-organellar communication. In pathogen-challenged Nicotiana benthamiana, stromule formation is driven by kinesins acting along microtubules. However, whether this mechanism also operates in unstressed plants or in other species remains unknown. Using systematic genetic analysis of all class XI myosin genes and quantitative time-lapse imaging in Arabidopsis (Arabidopsis thaliana), we show that basal stromule formation is predominantly myosin dependent through two distinct mechanisms. Myosin XI-I generates most stromules by driving oscillatory nuclear movement that mechanically pulls stromules from stationary plastids within the approx. 8m wide stromule-promoting zone. Myosins XI-K, XI-1, and XI-2 additionally drive a movement-independent, nucleus-associated stromule subpopulation within the same zone, establishing two mechanistically distinct pathways. We propose that movement-coupled stromules function as dynamic tethers maintaining plastid-nucleus proximity during nuclear oscillations, while movement-independent stromules expand the plastid membrane area within the perinuclear space, increasing the potential contact surface with the nucleus and surrounding environment. Together, both populations are proposed to enhance signal and metabolite exchange at the plastid-nucleus interface. These findings indicate that stromule biogenesis is both mechanistically and functionally context dependent, challenging the universality of the kinesin-microtubule model across plant species and physiological states.