In vivo analysis of interactions between GFP-labeled microfilaments and plastid stromules

BACKGROUND: Plastid stromules are stroma-filled tubules that extend from the surface of plastids in higher plants and allow the exchange of protein molecules between plastids. These structures are highly dynamic; stromules change both their shape and position in the cytoplasm very rapidly. Previous studies with microfilament inhibitors indicated that stromule shape and movement are dependent on the actin cytoskeleton. To learn more about the nature of the interactions of stromules and the cytoskeleton, we imaged fluorescently-labeled microfilaments and plastids. RESULTS: We have used Arabidopsis thaliana plants expressing green fluorescent protein fused to the human actin-binding protein talin to observe microfilaments and their relationship to stromules in vivo. Microfilaments were observed in close contact with stromules and plastid bodies of hypocotyl epidermis. Time-lapse confocal microscopy revealed that microfilament rearrangements were associated with changes in plastid and stromule morphology and position. We also observed close interactions between mitochondria and stromules in double-labeled cells. CONCLUSION: Our results indicate a correlation between the rearrangement of microfilaments and changes in the shape and position of plastids and stromules. Stromules interact with microfilaments that may also be utilized by mitochondria and other organelles. The interaction of microfilaments and plastids is likely to be mediated by actin-binding proteins on the plastid envelope membrane.