Arabidopsis PIN2 protein directs transport of the phytohormone auxin from the root tip into the root elongation zone. Variation in hormone transport, which depends on a delicate interplay between PIN2 sorting to and from polar plasma membrane domains, determines root growth. By employing a constitutively degraded version of PIN2, we identify brassinolides as antagonists of PIN2 endocytosis. This response does not require de novo protein synthesis, but involves early events in canonical brassinolide signaling. Brassinolide-controlled adjustments in PIN2 sorting and intracellular distribution governs formation of a lateral PIN2 gradient in gravistimulated roots, coinciding with adjustments in auxin signaling and directional root growth. Strikingly, simulations indicate that PIN2 gradient formation is no prerequisite for root bending but rather dampens asymmetric auxin flow and signaling. Crosstalk between brassinolide signaling and endocytic PIN2 sorting, thus, appears essential for determining the rate of gravity-induced root curvature via attenuation of differential cell elongation.
- MeSH
- Arabidopsis drug effects metabolism MeSH
- Biological Transport drug effects MeSH
- Brassinosteroids metabolism pharmacology MeSH
- Endocytosis drug effects MeSH
- Gravitropism drug effects physiology MeSH
- Plant Roots drug effects metabolism MeSH
- Indoleacetic Acids metabolism MeSH
- Meristem drug effects metabolism MeSH
- Arabidopsis Proteins metabolism MeSH
- Plant Growth Regulators metabolism pharmacology MeSH
- Signal Transduction MeSH
- Steroids, Heterocyclic metabolism pharmacology MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
As non-photosynthesizing organs, roots are dependent on diffusion of oxygen from the external environment and, in some instances, from the shoot for their aerobic metabolism. Establishment of hypoxic niches in the developing tissues of plants has been postulated as a consequence of insufficient diffusion of oxygen to satisfy the demands throughout development. Here, we report that such niches are established at specific stages of lateral root primordia development in Arabidopsis thaliana grown under aerobic conditions. Using gain- and loss-of-function mutants, we show that ERF-VII transcription factors, which mediate hypoxic responses, control root architecture by acting in cells with a high level of auxin signaling. ERF-VIIs repress the expression of the auxin-induced genes LBD16, LBD18, and PUCHI, which are essential for lateral root development, by binding to their promoters. Our results support a model in which the establishment of hypoxic niches in the developing lateral root primordia contributes to the shutting down of key auxin-induced genes and regulates the production of lateral roots.
- MeSH
- Arabidopsis cytology genetics metabolism MeSH
- Cell Hypoxia MeSH
- Plant Roots cytology MeSH
- Indoleacetic Acids metabolism MeSH
- Arabidopsis Proteins metabolism MeSH
- Gene Expression Regulation, Plant MeSH
- Signal Transduction * MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
The dual-affinity nitrate transceptor NITRATE TRANSPORTER1.1 (NRT1.1) has two modes of transport and signaling, governed by Thr-101 (T101) phosphorylation. NRT1.1 regulates lateral root (LR) development by modulating nitrate-dependent basipetal auxin export and nitrate-mediated signal transduction. Here, using the Arabidopsis (Arabidopsis thaliana) NRT1.1T101D phosphomimetic and NRT1.1T101A nonphosphorylatable mutants, we found that the phosphorylation state of NRT1.1 plays a key role in NRT1.1 function during LR development. Single-particle tracking revealed that phosphorylation affected NRT1.1 spatiotemporal dynamics. The phosphomimetic NRT1.1T101D form showed fast lateral mobility and membrane partitioning that facilitated auxin flux under low-nitrate conditions. By contrast, nonphosphorylatable NRT1.1T101A showed low lateral mobility and oligomerized at the plasma membrane (PM), where it induced endocytosis via the clathrin-mediated endocytosis and microdomain-mediated endocytosis pathways under high-nitrate conditions. These behaviors promoted LR development by suppressing NRT1.1-controlled auxin transport on the PM and stimulating Ca2+-ARABIDOPSIS NITRATE REGULATED1 signaling from the endosome.
- MeSH
- Arabidopsis genetics growth & development metabolism MeSH
- Nitrates metabolism MeSH
- Phosphorylation MeSH
- Plant Roots growth & development MeSH
- Indoleacetic Acids metabolism MeSH
- Arabidopsis Proteins metabolism MeSH
- Anion Transport Proteins genetics metabolism MeSH
- Plant Proteins genetics metabolism MeSH
- Transcription Factors metabolism MeSH
- Calcium Signaling MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
In Arabidopsis thaliana, canonical auxin-dependent gene regulation is mediated by 23 transcription factors from the AUXIN RESPONSE FACTOR (ARF) family that interact with auxin/indole acetic acid repressors (Aux/IAAs), which themselves form co-receptor complexes with one of six TRANSPORT INHIBITOR1/AUXIN-SIGNALLING F-BOX (TIR1/AFB) proteins. Different combinations of co-receptors drive specific sensing outputs, allowing auxin to control a myriad of processes. ARF6 and ARF8 are positive regulators of adventitious root initiation upstream of jasmonate, but the exact auxin co-receptor complexes controlling the transcriptional activity of these proteins has remained unknown. Here, using loss-of-function mutants we show that three Aux/IAA genes, IAA6, IAA9, and IAA17, act additively in the control of adventitious root (AR) initiation. These three IAA proteins interact with ARF6 and/or ARF8 and likely repress their activity in AR development. We show that TIR1 and AFB2 are positive regulators of AR formation and TIR1 plays a dual role in the control of jasmonic acid (JA) biosynthesis and conjugation, as several JA biosynthesis genes are up-regulated in the tir1-1 mutant. These results lead us to propose that in the presence of auxin, TIR1 and AFB2 form specific sensing complexes with IAA6, IAA9, and/or IAA17 to modulate JA homeostasis and control AR initiation.
- MeSH
- Arabidopsis cytology genetics growth & development metabolism MeSH
- F-Box Proteins metabolism MeSH
- Hypocotyl metabolism MeSH
- Plant Roots growth & development MeSH
- Indoleacetic Acids metabolism MeSH
- Arabidopsis Proteins metabolism MeSH
- Receptors, Cell Surface metabolism MeSH
- Gene Expression Regulation, Plant MeSH
- Signal Transduction * MeSH
- Protein Stability MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
The plant hormone auxin and its directional transport are known to play a crucial role in defining the embryonic axis and subsequent development of the body plan. Although the role of PIN auxin efflux transporters has been clearly assigned during embryonic shoot and root specification, the role of the auxin influx carriers AUX1 and LIKE-AUX1 (LAX) proteins is not well established. Here, we used chemical and genetic tools on Brassica napus microspore-derived embryos and Arabidopsis thaliana zygotic embryos, and demonstrate that AUX1, LAX1 and LAX2 are required for both shoot and root pole formation, in concert with PIN efflux carriers. Furthermore, we uncovered a positive-feedback loop between MONOPTEROS (ARF5)-dependent auxin signalling and auxin transport. This MONOPTEROS-dependent transcriptional regulation of auxin influx (AUX1, LAX1 and LAX2) and auxin efflux (PIN1 and PIN4) carriers by MONOPTEROS helps to maintain proper auxin transport to the root tip. These results indicate that auxin-dependent cell specification during embryo development requires balanced auxin transport involving both influx and efflux mechanisms, and that this transport is maintained by a positive transcriptional feedback on auxin signalling.
- MeSH
- Arabidopsis embryology genetics metabolism MeSH
- Biological Transport genetics physiology MeSH
- Brassica napus embryology genetics metabolism MeSH
- Indoleacetic Acids metabolism MeSH
- Plant Proteins metabolism MeSH
- Seeds cytology metabolism MeSH
- Signal Transduction genetics physiology MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
Polar auxin transport plays a pivotal role in plant growth and development. PIN-FORMED (PIN) auxin efflux carriers regulate directional auxin movement by establishing local auxin maxima, minima, and gradients that drive multiple developmental processes and responses to environmental signals. Auxin has been proposed to modulate its own transport by regulating subcellular PIN trafficking via processes such as clathrin-mediated PIN endocytosis and constitutive recycling. Here, we further investigated the mechanisms by which auxin affects PIN trafficking by screening auxin analogs and identified pinstatic acid (PISA) as a positive modulator of polar auxin transport in Arabidopsis (Arabidopsis thaliana). PISA had an auxin-like effect on hypocotyl elongation and adventitious root formation via positive regulation of auxin transport. PISA did not activate SCFTIR1/AFB signaling and yet induced PIN accumulation at the cell surface by inhibiting PIN internalization from the plasma membrane. This work demonstrates PISA to be a promising chemical tool to dissect the regulatory mechanisms behind subcellular PIN trafficking and auxin transport.
- MeSH
- Arabidopsis drug effects metabolism MeSH
- Biological Transport drug effects MeSH
- Cell Membrane drug effects metabolism MeSH
- Endocytosis * drug effects MeSH
- Phenotype MeSH
- Phenylacetates pharmacology MeSH
- Gravitropism drug effects MeSH
- Hypocotyl drug effects growth & development MeSH
- Plant Roots drug effects growth & development MeSH
- Indoleacetic Acids metabolism MeSH
- Arabidopsis Proteins metabolism MeSH
- Signal Transduction MeSH
- Plant Shoots metabolism MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
Being sessile organisms, plants have evolved mechanisms allowing them to control their growth and development in response to environmental changes. This occurs by means of complex interacting signalling networks that integrate diverse environmental cues into co-ordinated and highly regulated responses. Auxin is an essential phytohormone that functions as a signalling molecule, driving both growth and developmental processes. It is involved in numerous biological processes ranging from control of cell expansion and cell division to tissue specification, embryogenesis, and organ development. All these processes require the formation of auxin gradients established and maintained through the combined processes of biosynthesis, metabolism, and inter- and intracellular directional transport. Environmental conditions can profoundly affect the plant developmental programme, and the co-ordinated shoot and root growth ought to be fine-tuned to environmental challenges such as temperature, light, and nutrient and water content. The key role of auxin as an integrator of environmental signals has become clear in recent years, and emerging evidence implicates auxin biosynthesis as an essential component of the overall mechanisms of plants tolerance to stress. In this review, we provide an account of auxin's role as an integrator of environmental signals and, in particular, we highlight the effect of these signals on the control of auxin production.
The long-standing Acid Growth Theory of plant cell elongation posits that auxin promotes cell elongation by stimulating cell wall acidification and thus expansin action. To date, the paucity of pertinent genetic materials has precluded thorough analysis of the importance of this concept in roots. The recent isolation of mutants of the model grass species Brachypodium distachyon with dramatically enhanced root cell elongation due to increased cellular auxin levels has allowed us to address this question. We found that the primary transcriptomic effect associated with elevated steady state auxin concentration in elongating root cells is upregulation of cell wall remodeling factors, notably expansins, while plant hormone signaling pathways maintain remarkable homeostasis. These changes are specifically accompanied by reduced cell wall arabinogalactan complexity but not by increased proton excretion. On the contrary, we observed a tendency for decreased rather than increased proton extrusion from root elongation zones with higher cellular auxin levels. Moreover, similar to Brachypodium, root cell elongation is, in general, robustly buffered against external pH fluctuation in Arabidopsis thaliana However, forced acidification through artificial proton pump activation inhibits root cell elongation. Thus, the interplay between auxin, proton pump activation, and expansin action may be more flexible in roots than in shoots.
Identification of mutants with impairments in auxin biosynthesis and dynamics by forward genetic screening is hindered by the complexity, redundancy and necessity of the pathways involved. Furthermore, although a few auxin-deficient mutants have been recently identified by screening for altered responses to shade, ethylene, N-1-naphthylphthalamic acid (NPA) or cytokinin (CK), there is still a lack of robust markers for systematically isolating such mutants. We hypothesized that a potentially suitable phenotypic marker is root curling induced by CK, as observed in the auxin biosynthesis mutant CK-induced root curling 1 / tryptophan aminotransferase of Arabidopsis 1 (ckrc1/taa1). Phenotypic observations, genetic analyses and biochemical complementation tests of Arabidopsis seedlings displaying the trait in large-scale genetic screens showed that it can facilitate isolation of mutants with perturbations in auxin biosynthesis, transport and signaling. However, unlike transport/signaling mutants, the curled (or wavy) root phenotypes of auxin-deficient mutants were significantly induced by CKs and could be rescued by exogenous auxins. Mutants allelic to several known auxin biosynthesis mutants were re-isolated, but several new classes of auxin-deficient mutants were also isolated. The findings show that CK-induced root curling provides an effective marker for discovering genes involved in auxin biosynthesis or homeostasis.
- MeSH
- Arabidopsis enzymology genetics MeSH
- Biological Transport drug effects MeSH
- Cytokinins metabolism MeSH
- Phenotype MeSH
- Plant Roots enzymology genetics growth & development MeSH
- Indoleacetic Acids metabolism pharmacology MeSH
- Mutation MeSH
- Arabidopsis Proteins genetics metabolism MeSH
- Plant Growth Regulators biosynthesis MeSH
- Seedlings drug effects growth & development metabolism MeSH
- Signal Transduction drug effects MeSH
- Tryptophan Transaminase genetics metabolism MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
