Here, we provide a novel mechanistic framework for cell polarization during auxin-driven plant development that combines intracellular auxin signaling for regulation of expression of PINFORMED (PIN) auxin efflux transporters and the theoretical assumption of extracellular auxin signaling for regulation of PIN subcellular dynamics.The competitive utilization of auxin signaling component in the apoplast might account for the elusive mechanism for cell-to-cell communication for tissue polarization.Computer model simulations faithfully and robustly recapitulate experimentally observed patterns of tissue polarity and asymmetric auxin distribution during formation and regeneration of vascular systems, and during the competitive regulation of shoot branching by apical dominance.Our model generated new predictions that could be experimentally validated, highlighting a mechanistically conceivable explanation for the PIN polarization and canalization of the auxin flow in plants.
A key question of developmental biology relates to a fundamental issue in cell and tissue polarities, namely, how an individual cell in a polarized tissue senses the polarities of its neighbors and its position within tissue. In plant development, this issue is of pronounced importance, because plants have a remarkable ability to redefine cell and tissue polarities in different developmental programs, such as embryogenesis, postembryonic organogenesis, vascular tissue formation, and tissue regeneration (Kleine-Vehn and Friml, 2008).
A polar, cell-to-cell transport of the small signaling molecule auxin in conjunction with local auxin biosynthesis determines auxin gradients during embryonic and postembryonic development, giving positional cues for primordia formation, organ patterning, and tropistic growth (Friml et al, 2002; Benková et al, 2003; Reinhardt et al, 2003; Heisler et al, 2005; Scarpella et al, 2006; Dubrovsky et al, 2008). Over the past decades, theoretical models proposed that auxin acts as a polarizing cue in the center of a positive feedback mechanisms for auxin transport that has a key role in synchronized polarity rearrangements. However, the mechanistic basis for such a feedback loop between auxin and its own transport remains to a large extent elusive.
The direction of auxin transport largely depends on the polar subcellular localization of PINFORMED (PIN) proteins at the plasma membrane (Petrášek et al, 2006; Wiśniewska et al, 2006). These proteins recycle between the plasma membrane and intracellular endosomal compartments (Geldner et al, 2001; Dhonukshe et al, 2007), and their recycling modulates PIN-dependent auxin efflux rates and enable rapid changes in PIN polarity (Dubrovsky et al, 2008; Kleine-Vehn et al, 2008a). Nevertheless, the molecular basis for PIN polarization in plants remains unknown.
To gain new mechanistic insights in the hypothetical feedback mechanisms governing PIN polarization, several theoretical studies (Mitchison, 1980; Sachs, 1981; Rolland-Lagan and Prusinkiewicz, 2005; Jönsson et al, 2006; Smith et al, 2006; Merks et al, 2007; Bayer et al, 2009; Kramer, 2009) have been carried out. These models suggest that auxin promotes its own transport by modulating the amount of PIN proteins at the plasma membrane by incorporating either not yet identified flux gradient-based component (Mitchison, 1980; Rolland-Lagan and Prusinkiewicz, 2005; Bayer et al, 2009; Kramer, 2009) or an unknown short-range intercellular signal-transmitting auxin concentrations of its direct neighbors (Jönsson et al, 2006; Smith et al, 2006; Merks et al, 2007; Bayer et al, 2009; Sahlin et al, 2009).
Here, we propose a feedback driven, biologically plausible model for PIN polarization and auxin transport that introduces the combination of intracellular and extracellular auxin signaling pathways as a unified approach for tissue polarization in plants. Our computer model is based on chemiosmotic hypothesis (Goldsmith et al, 1981; Figure 1A) and integrates up-to-date experimental data, such as auxin feedback on PIN expression (Peer et al, 2004; Heisler et al, 2005) via a nuclear auxin signaling pathway (Chapman and Estelle, 2009; Figure 1B), auxin carrier recycling auxin (Dubrovsky et al, 2008; Kleine-Vehn et al, 2008a; Figure 1C), and auxin feedback on PIN endocytosis (Paciorek et al, 2005) via novel hypothetical, yet plausible, assumption of extracellular auxin perception (Figure 1D).
The heart of our extracellular receptor-based polarization (ERP) mechanism is the competitive utilization of auxin receptors in the intercellular space that allows a direct and simple cell-to-cell communication scheme. In our model, auxin binds to its extracellular receptor in the concentration-dependent manner and induces signal to modulate PIN protein abundance at the plasma membrane (Figure 1D). The direct mode of the signal transfer involves temporal immobilization of recruited receptors to the plasma membrane, which is reflected by reduced diffusion of receptors involved in auxin signaling (Figure 1D). This competitive utilization mechanism enables cell-to-cell communication in our model, leading to receptor enrichment at the site of higher auxin concentration (Figure 1D). The PIN polarization and polar auxin transport in our model both depend on and contribute to the establishment of differential auxin signaling in the cell wall. This feedback loop leads ultimately to the alignment of PIN polarization within a tissue.
We demonstrated the plausibility of the ERP model for various processes, including de novo vascularization, venation patterning, and tissue regeneration in computer simulations performed with only minimal initial assumptions, a discrete auxin source, and a distal sink. The ERP model reproduces the very detailed PIN polarization events that occur during primary vein initiation (Scarpella et al, 2006), such as basal PIN1 polarity in provascular cells, transient adverse PIN1 polarization in neighboring cells during the alignment of tissue polarization, and inner-lateral polarity displayed by the tissues surrounding a conductive auxin channel (Figure 3). Additionally, the ERP model generates high auxin concentration and high auxin flux simultaneously in emerging veins, revising the classical canalization models (Mitchison, 1980; Rolland-Lagan and Prusinkiewicz, 2005). Importantly, all our model simulations support the claim that the ERP model represents the first single approach that faithfully reproduces PIN polarization, both with the auxin gradient (basal PIN1 polarity in provascular cells) and against the auxin gradient (transient adverse PIN1 polarization in neighboring cells surrounding the provascular bundle), as well as producing the corresponding auxin distribution patterns during auxin canalization.
The proposed model introduces the extracellular auxin signaling pathway, which is crucial to account for coordinated PIN polarization and auxin distribution during venation patterning in plants. The putative candidate for extracellular auxin receptor is auxin-binding protein 1 (ABP1), which resides in the lumen of the endoplasmic reticulum and is secreted to the cell wall (Napier et al, 2002; Tromas et al, 2009) where it is physiologically active (Leblanc et al, 1999; Steffens et al, 2001). Additionally, auxin inhibits clathrin-dependent PIN internalization via binding to ABP1 (Robert et al, 2010). Thus, we speculate that the extracellular fraction of ABP1 (or additionally yet to be identified ABPs) could correspond to the common pool of extracellular auxin receptors in the ERP model. A future challenge will be to test whether the ERP model unifies complex PIN polarization and auxin distribution patterns in embryogenesis, root system maintenance, and de novo organ formation.
Plant development is exceptionally flexible as manifested by its potential for organogenesis and regeneration, which are processes involving rearrangements of tissue polarities. Fundamental questions concern how individual cells can polarize in a coordinated manner to integrate into the multicellular context. In canalization models, the signaling molecule auxin acts as a polarizing cue, and feedback on the intercellular auxin flow is key for synchronized polarity rearrangements. We provide a novel mechanistic framework for canalization, based on up-to-date experimental data and minimal, biologically plausible assumptions. Our model combines the intracellular auxin signaling for expression of PINFORMED (PIN) auxin transporters and the theoretical postulation of extracellular auxin signaling for modulation of PIN subcellular dynamics. Computer simulations faithfully and robustly recapitulated the experimentally observed patterns of tissue polarity and asymmetric auxin distribution during formation and regeneration of vascular systems and during the competitive regulation of shoot branching by apical dominance. Additionally, our model generated new predictions that could be experimentally validated, highlighting a mechanistically conceivable explanation for the PIN polarization and canalization of the auxin flow in plants.