Energy is essential for all cellular functions in a living organism. How cells coordinate their physiological processes with energy status and availability is thus an important question. The turnover of actin cytoskeleton between its monomeric and filamentous forms is a major energy drain in eukaryotic cells. However, how actin dynamics are regulated by ATP levels remain largely unknown in plant cells. Here, we observed that seedlings with impaired functions of target of rapamycin complex 1 (TORC1), either by mutation of the key component, RAPTOR1B , or inhibition of TOR activity by specific inhibitors, displayed reduced sensitivity to actin cytoskeleton disruptors compared to their controls. Consistently, actin filament dynamics, but not organization, were suppressed in TORC1-impaired cells. Subcellular localization analysis and quantification of ATP concentration demonstrated that RAPTOR1B localized at cytoplasm and mitochondria and that ATP levels were significantly reduced in TORC1-impaired plants. Further pharmacologic experiments showed that the inhibition of mitochondrial functions led to phenotypes mimicking those observed in raptor1b mutants at the level of both plant growth and actin dynamics. Exogenous feeding of adenine could partially restore ATP levels and actin dynamics in TORC1-deficient plants. Thus, these data support an important role for TORC1 in coordinating ATP homeostasis and actin dynamics in plant cells.

Pollen germination and pollen tube growth are important biological events in the sexual reproduction of higher plants, during which a large number of vesicle trafficking and membrane fusion events occur. When secretory vesicles are transported via the F-actin network in proximity to the apex of the pollen tube, the secretory vesicles are tethered and fused to the plasma membrane by tethering factors and SNARE proteins, respectively. The coupling and uncoupling between the vesicle membrane and plasma membrane are also regulated by dynamic cytoskeleton, proteins, and signaling molecules, including small G proteins, calcium, and PIP2. In this review, we focus on the current knowledge regarding secretory vesicle delivery, tethering, and fusion during pollen germination and tube growth and summarize the progress in research on how regulators and signaling molecules participate in the above processes.

Summary: All living organisms are subjected to mechanical forces at all times. It has been reported that mechanics regulate many key cellular processes, including cell polarity establishment, cell division and gene expression, as a physical signal in both animal and plant development. Plant cells are exposed to several types of mechanical stresses, ranging from turgor‐driven tensile stresses, mechanical force modified by heterogeneous growth directions and rates between neighbouring cells, to forces from the environment such as wind and rain, for which they have developed adaptive mechanisms. Increasing evidence has revealed that mechanical stresses markedly influence the alignment of cortical microtubules (CMTs) in plant cells, among other effects. CMTs are able to reorient in response to mechanical stresses at both the single‐cell and tissue levels and always align with the maximal tensile stress direction. In this review, we discussed the known and potential molecules and pathways involved in the regulation of CMTs by mechanical stresses. We also summarized the available techniques that have allowed for mechanical perturbation. Finally, we highlighted several key questions remaining to be addressed in this emerging field.

Phosphatidylinositol 4,5‐bisphosphate (PI(4,5)P₂) is known to be an instrumental anionic phospholipid in governing pollen germination and pollen tube growth. However, the precise functions and regulatory mechanisms of PI(4,5)P₂ in pollen polarity establishment and germination remain poorly understood. Our previous studies demonstrated the pivotal involvement of Arabidopsis formin homology 5 (AtFH5)‐dependent vesicle trafficking in polarity establishment of pollen. Here, we observed that PI(4,5)P₂ accumulated and oscillated at the prospective germination site, a process closely associated with the rotational movement of AtFH5‐labeled vesicles. Disruption of the mobility of AtFH5‐labeled vesicles, either through AtFH5 mutation or pharmacological treatment, significantly perturbed the accumulation of PI(4,5)P₂ at the plasma membrane. Subcellular localization and genetic analysis revealed that two phosphatidylinositol 4‐phosphate 5‐kinases, AtPIP5K1 and AtPIP5K4, are essential for PI(4,5)P₂ oscillation at the germination site prior to pollen germination. Furthermore, we found that the dynamics of AtPIP5K4 depended on the mobility of AtFH5‐labeled vesicles and reduced PI(4,5)P₂ in turn disturbed the attachment of AtFH5‐labeled secretory vesicles to the plasma membrane. In conclusion, these findings collectively highlight the reciprocal regulation of AtFH5‐labeled secretory vesicles and PI(4,5)P₂ oscillations at the plasma membrane, providing critical insights into the molecular mechanism underlying polarity establishment during pollen germination.

Actin cytoskeleton drives the targeted transport of cell wall components to sustain the tip growth of pollen tubes for double fertilization; however, the underlying mechanism remains largely unknown. Arabidopsis formin 5 (AtFH5), an actin-nucleating protein, localizes at secretory vesicles and mediates actin polymerization-based vesicle trafficking in pollen. Here, we demonstrate that AtFH5 determines the recruitment and transport of cell wall components in AtFH5-labeled vesicles during the tip growth of pollen tubes. Through a screen of interacting proteins of AtFH5, we identify many cell wall–related proteins, with arabinogalactan protein 23 (AGP23) occupying the highest frequency. AtFH5 interacts with AGP23 via its N-terminal extracellular domain (ECD) and jointly regulate the pollen germination and tube growth process. Further observations reveal that AGP23 co-localizes with AtFH5 at moving vesicles, germination sites, and pollen tube tips, suggesting that AGP23 is delivered by AtFH5-labeled vesicles. Deletion of the ECD of AtFH5 interrupts the dynamic localization and cell-wall connection of AGP23 in pollen grains and tubes. Cytological and genetic evidence shows that AGP23 and AtFH5 work in the same pathway to modulate cell wall composition. Together, our data uncover a role of formin in directing the sorting and deposition of cell wall components via secretory vesicle trafficking during pollen germination and tube growth.

The plant cytoskeleton is an intricate network composed of actin filaments and microtubules. The cytoskeleton undergoes continuous dynamic changes that provide the basis for rapidly responding to intrinsic and extrinsic stimuli, including mechanical stress. Microtubules can respond to alterations of mechanical stress and reorient along the direction of maximal tensile stress in plant cells. The cytoskeleton can also generate driving force for cytoplasmic streaming, organelle movement, and vesicle transportation. In this review, we discuss the progress of how the plant cytoskeleton responds to mechanical stress. We also summarize the roles of the cytoskeleton in generating force that drive organelles and nuclear transportation in plant cells. Finally, some hypotheses concerning the link between the roles of the cytoskeleton in force response and organelle movement, as well as several key questions that remain to be addressed in the field, are highlighted.