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Plants produce a rich diversity of biological forms, and the diversity of leaves is especially notable. Mechanisms of leaf morphogenesis have been studied in the past two decades, with a growing focus on the interactive roles of mechanics in recent years. Growth of plant organs involves feedback by mechanical stress: growth induces stress, and stress affects growth and morphogenesis. Although much attention has been given to potential stress-sensing mechanisms and cellular responses, the mechanical principles guiding morphogenesis have not been well understood. Here we synthesize the overarching roles of mechanics and mechanical stress in multilevel and multiple stages of leaf morphogenesis, encompassing leaf primordium initiation, phyllotaxis and venation patterning, and the establishment of complex mature leaf shapes. Moreover, the roles of mechanics at multiscale levels, from subcellular cytoskeletal molecules to single cells to tissues at the organ scale, are articulated. By highlighting the role of mechanical buckling in the formation of three-dimensional leaf shapes, this review integrates the perspectives of mechanics and biology to provide broader insights into the mechanobiology of leaf morphogenesis. Mechanical regulation of plant growth is rapidly gaining the interest of researchers. This review details how mechanics takes part in leaf morphogenesis during the initiation of leaf primordia, emergence of phyllotaxis patterns, and development of complex leaf shapes. By highlighting the role of mechanical buckling in leaf shape formation, this review integrates perspectives of mechanics and biology to provide insights into plant mechanobiology. [ABSTRACT FROM AUTHOR]

Delivery of proteins to the plasma membrane occurs via secretion, which requires tethering, docking, priming, and fusion of vesicles. In yeast and mammalian cells, an evolutionarily conserved RAB GTPase activation cascade functions together with the exocyst and SNARE proteins to coordinate vesicle transport with fusion at the plasma membrane. However, it is unclear whether this is the case in plants. In this study, we show that the small GTPase RABA2a recruits and interacts with the VAMP721/722-SYP121-SNAP33 SNARE ternary complex for membrane fusion. Through immunoprecipitation coupled with mass spectrometry analysis followed by the validatation with a series of biochemical assays, we identified the SNARE proteins VAMP721 and SYP121 as the interactors and downstream effectors of RABA2a. Further expreiments showed that RABA2a interacts with all members of the SNARE complex in its GTP-bound form and modulates the assembly of the VAMP721/722-SYP121-SNAP33 SNARE ternary complex. Intriguingly, we did not observe the interaction of the exocyst subunits with either RABA2a or theSNARE proteins in several different experiments. Neither RABA2a inactivation affects the subcellular localization or assembly of the exocystnor the exocyst subunit mutant exo84b shows the disrupted RABA2a-SNARE association or SNARE assembly, suggesting that the RABA2a-SNARE- and exocyst-mediated secretory pathways are largely independent. Consistently, our live imaging experiments reveal that the two sets of proteins follow non-overlapping trafficking routes, and genetic and cell biologyanalyses indicate that the two pathways select different cargos. Finally, we demonstrate that the plant-specific RABA2a-SNARE pathway is essential for the maintenance of potassium homeostasis in Arabisopsis seedlings. Collectively, our findings imply that higher plants might have generated different endomembrane sorting pathways during evolution and may enable the highly conserved endomembrane proteins to participate in plant-specific trafficking mechanisms for adaptation to the changing environment. Classic cell biology studies suggest that the exocyst is required to anchor secretory vesicles to the PM prior to SNARE complex-mediated membrane fusion. In this study, the authors unexpectedly found that the small GTPase RABA2a interacts with the SNARE complex for membrane fusion, bypassing the exocyst. Further analyses provide multiple lines of evidence supporting that the RABA2a-SNARE- and exocyst-mediated secretory pathways are largely independent. [ABSTRACT FROM AUTHOR]

The first asymmetric meiotic cell divisions in mouse oocytes are driven by formin 2 (FMN2)-nucleated actin polymerization around the spindle. In this study, we investigated how FMN2 is recruited to the spindle peripheral ER and how its activity is regulated in mouse meiosis I (MI) oocytes. We show that this process is regulated by the Ran GTPase, a conserved mediator of chromatin signal, and the ER-associated protein VAPA. FMN2 contains a nuclear localization sequence (NLS) within a domain (SLD) previously shown to be required for FMN2 localization to the spindle periphery. FMN2 NLS is bound to the importin α1/β complex, and the disruption of this interaction by RanGTP is required for FMN2 accumulation in the area proximal to the chromatin and the MI spindle. The importin-free FMN2 is then recruited to the surface of ER around the spindle through the binding of the SLD with the ER-membrane protein VAPA. We further show that FMN2 is autoinhibited through an intramolecular interaction between the SLD with the C-terminal formin homology 2 (FH2) domain that nucleates actin filaments. VAPA binding to SLD relieves the autoinhibition of FMN2, leading to localized actin polymerization. This dual control of formin-mediated actin assembly allows actin polymerization to initiate the movement of the meiotic spindle toward the cortex, an essential step in the maturation of the mammalian female gamete. [Display omitted] • FMN2 is an NLS-containing formin protein bound to importin α1/β complex • Ran GTPase regulates FMN2 recruitment to spindle periphery during oocyte MI • Importin dissociation by Ran allows VAPA to anchor FMN2 to ER membrane • VAPA binding relieves FMN2 autoinhibition to nucleate actin assembly The first asymmetric meiotic division is essential for the formation of a functional mature female gamete. Wang et al. demonstrate that dual control of formin 2-mediated actin assembly by Ran GTPase signaling and an ER-membrane protein restricts actin polymerization from the ER surface at the spindle periphery. This initiates the meiotic spindle migration from the center of the oocyte to the cortex. [ABSTRACT FROM AUTHOR]

Summary Profilin functions with formin in actin assembly, a process that regulates multiple aspects of plant development and immune responses. High-level eukaryotes contain multiple isoforms of profilin, formin, and actin, whose partner-specific interactions in actin assembly are not completely understood in plant development and defense responses. To examine the functionally distinct interactions between profilin and formin, we studied all five Arabidopsis profilins and their interactions with formin by using both in vitro biochemical and in vivo cell biology approaches. Unexpectedly, we found a previously undescribed negative regulatory function of AtPRF3 in AtFH1-mediated actin polymerization. The N-terminal 37 residues of AtPRF3 were identified to play a predominant role in inhibiting formin-mediated actin nucleation via their high affinity for the formin polyproline region and their triggering of the oligomerization of AtPRF3. Both in vivo and in vitro mechanistic studies of AtPRF3 revealed a universal mechanism in which the weak interaction between profilin and formin positively regulates actin assembly by ensuring rapid recycling of profilin, whereas profilin oligomerization negatively regulates actin polymerization. Upon recognition of the pathogen-associated molecular pattern, the gene transcription and protein degradation of AtPRF3 are modulated for actin assembly during plant innate immunity. The prf3 Arabidopsis plants show higher sensitivity to the bacterial flagellum peptide in both the plant growth and ROS responses. These findings demonstrate a profilin-mediated actin assembly mechanism underlying the plant immune responses. [ABSTRACT FROM AUTHOR]

Coronins play critical roles in actin network formation. The diverse functions of coronins are regulated by the structured N-terminal β propeller and the C-terminal coiled coil (CC). However, less is known about a middle "unique region" (UR), which is an intrinsically disordered region (IDR). The UR/IDR is an evolutionarily conserved signature in the coronin family. By integrating biochemical and cell biology experiments, coarse-grained simulations, and protein engineering, we find that the IDR optimizes the biochemical activities of coronins in vivo and in vitro. The budding yeast coronin IDR plays essential roles in regulating Crn1 activity by fine-tuning CC oligomerization and maintaining Crn1 as a tetramer. The IDR-guided optimization of Crn1 oligomerization is critical for F-actin cross-linking and regulation of Arp2/3-mediated actin polymerization. The final oligomerization status and homogeneity of Crn1 are contributed by three examined factors: helix packing, the energy landscape of the CC, and the length and molecular grammar of the IDR., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.)

Plants respond to bacterial infection acutely with actin remodeling during plant immune responses. The mechanisms by which bacterial virulence factors (VFs) modulate plant actin polymerization remain enigmatic. Here, we show that plant-type-I formin serves as the molecular sensor for actin remodeling in response to two bacterial VFs: Xanthomonas campestris pv. campestris (Xcc) diffusible signal factor (DSF), and pathogen-associated molecular pattern (PAMP) flagellin in pattern-triggered immunity (PTI). Both VFs regulate actin assembly by tuning the clustering and nucleation activity of formin on the plasma membrane (PM) at the nano-sized scale. By being integrated within the cell-wall-PM-actin cytoskeleton (CW-PM-AC) continuum, the dynamic behavior and function of formins are highly dependent on each scaffold layer's composition within the CW-PM-AC continuum during both DSF and PTI signaling. Our results reveal a central mechanism for rapid actin remodeling during plant-bacteria interactions, in which bacterial signaling molecules fine tune plant formin nanoclustering in a host mechanical-structure-dependent manner., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2021 The Author(s). Published by Elsevier Inc. All rights reserved.)