Transmembrane auxin carrier systems – dynamic regulators of polar auxin transport

Recent investigations of the biochemistry, physiology and molecular genetics of polar auxin transport have greatly advanced our understanding of the process and of the part it plays in the regulation of development and in the responses of cells, tissues and organs to internal and external stimuli. The molecular and physiological characterization of mutants which exhibit lesions in polar auxin transport has led to the isolation and sequencing of genes which encode putative components of auxin carrier systems, or proteins which directly or indirectly regulate these systems. This work has revealed that specific auxin uptake and efflux carriers are coded not by single genes, but by whole families of genes, the expression of which is tissue or stimulus specific. Furthermore, evidence is accumulating rapidly that at least the auxin efflux carrier is a multi-component system consisting of both catalytic and regulatory subunits, including a separate phytotropin-binding protein. Other genes have been tentatively identified which code proteins that regulate the expression of genes coding auxin carrier components, or which regulate the intracellular traffic or activity of auxin carriers. Investigations of the turn-over and Golgi-mediated trafficking of auxin carrier proteins have revealed that essential components of at least the efflux carrier have a very short half-life in the plasma membrane and are replaced without the need for concurrent protein synthesis, leading to speculation that they might cycle between internal stores and the plasma membrane. The way is now clear for the development of specific molecular probes with which to investigate the intracellular transport and targeting of auxin carrier proteins.     

Function of the ubiquitin–proteosome pathway in auxin response

Proteolysis of important regulatory proteins by the ubiquitin–proteosome pathway is a key aspect of cellular regulation in eukaryotes. Genetic studies in Arabidopsis indicate that response to auxin depends on the function of proteins in this pathway. The auxin transport inhibitor resistant 1 (TIR1) protein is part of a ubiquitin–protein–ligase complex (E3), known as SKP1 CDC53 F-box^TIR1 (SCF^TIR1), that possibly directs ubiquitin-modification of protein regulators of the auxin response. In yeast, a similar E3 complex, SCF^CDC4, is regulated by conjugation of the ubiquitin-related protein Rub1 to the Cdc53 protein. In Arabidopsis, the auxin-resistant1 (AXR1) gene encodes a subunit of the RUB1-activating enzyme, the first enzyme in the RUB-conjugation pathway. Loss of AXR1 results in loss of auxin response. These results suggest a model in which RUB1 modification regulates the activity of SCF^TIR1, thereby directing the degradation of the repressors of the auxin response.     

Does ethylene induce elongation of the rice coleoptile through auxin?

Ethylene stimulated the elongation of intact rice (Oryza sativaL.) coleoptiles in which endogenous growth had been stoppedcompletely by decapitation and red light. p-Chlorophenoxyisobutyricacid slightly inhibited endogenous growth, but not the ethyleneinduced growth. Thus, ethylene could stimulate the elongationof coleoptiles in which the auxin level was considered to bevery low. ¹ Present address: Institute for Agricultural Research, TohokuUniversity, Katahira, Sendai 980, Japan. (Received February 16, 1979; )     

Is auxin the repressor signal of branch growth in apical control?

"Apical control" is the repression of branch growth by a higher dominating branch or shoot. There has been some confusion in the literature concerning the meaning and causal mechanisms of this correlative phenomenon with those of "apical dominance," which term is often used in a strict sense to connote the repression of the initiation of axillary bud outgrowth by an active shoot apex. Although the term "apical control" is most commonly employed with respect to woody species, this phenomenon also widely occurs in herbaceous plants. Because of the strong evidence for a role of auxin as a repressor signal in apical dominance and partly because of this lack of distinction in terminology, a similar role for auxin in apical control is often assumed in spite of the obvious acropetal auxin transport difficulty and the lack of direct evidence for the acropetal transport of any inhibitor influence. In the present study with the herbaceous Ipomoea nil, it has been clearly demonstrated that while exogenous auxin (1% NAA) strongly restores apical dominance in the Thimann-Skoog experiment, auxin treatments to decapitated dominant shoots do not, in any observable way, restore apical control in lower dominated branches. Hence, in this fast-growing species, the hypothesis for the role of auxin as a repressor signal for apical control is not supported.     

Halotropism requires phospholipase Dζ1-mediated modulation of cellular polarity of auxin transport carriers

Endocytosis and relocalization of auxin carriers represent important mechanisms for adaptive plant growth and developmental responses. Both root gravitropism and halotropism have been shown to be dependent on relocalization of auxin transporters. Following their homology to mammalian phospholipase Ds (PLDs), plant PLDζ-type enzymes are likely candidates to regulate auxin carrier endocytosis. We investigated root tropic responses for an Arabidopsis pldζ1-KO mutant and its effect on the dynamics of two auxin transporters during salt stress, that is, PIN2 and AUX1. We found altered root growth and halotropic and gravitropic responses in the absence of PLDζ1 and report a role for PLDζ1 in the polar localization of PIN2. Additionally, irrespective of the genetic background, salt stress induced changes in AUX1 polarity. Utilizing our previous computational model, we found that these novel salt-induced AUX1 changes contribute to halotropic auxin asymmetry. We also report the formation of “osmotic stress-induced membrane structures.” These large membrane structures are formed at the plasma membrane shortly after NaCl or sorbitol treatment and have a prolonged presence in a pldζ1 mutant. Taken together, these results show a crucial role for PLDζ1 in both ionic and osmotic stress-induced auxin carrier dynamics during salt stress.     

Regulation of seedling growth by ethylene and the ethylene–auxin crosstalk

Planta - This review highlights that the auxin gradient, established by local auxin biosynthesis and transport, can be controlled by ethylene, and steers seedling growth. A better understanding of...     

Identification of dehydrocostus lactone and 4-hydroxy-β-thujone as auxin polar transport inhibitors

The survey of naturally occurring of auxin polar transport regulators in Asteraceae was investigated using the radish (Raphanus sativus L.) hypocotyl bioassay established in this study. Significant auxin polar transport was observed when radiolabeled indole-3-acetic acid (IAA) was applied at the apical side of radish hypocotyl segments, but not when it was applied at the basal side of the segments. Almost no auxin polar transport was observed in radish hypocotyl segments treated with synthetic auxin polar transport inhibitors of N-(1-naphthyl)phthalamic acid (NPA) and 9-hydroxyfluorene-9-carboxylic acid (HFCA) at 0.5 μg/plant. 2,3,5-Triiodobenzoic acid (TIBA) at 0.5 μg/plant was less effective than NPA and HFCA, and p-chlorophenoxyisobutyric acid (PCIB) at 0.5 μg/plant had almost no effect on auxin polar transport in the radish hypocotyl bioassay. These results strongly suggest that the radish hypocotyl bioassay is suitable for the detection of bioassay-derived auxin polar transport regulators. Using the radish hypocotyl bioassay and physicochemical analyses, dehydrocostus lactone (decahydro-3,6,9-tris-methylene-azulenol(4,5-b)furan-2(3H)-one) and 4-hydroxy-β-thujone (4-hydroxy-4-methyl-1-(1-methylethyl)-bicyclo[3.1.0]hexan-3-one) were successfully identified as auxin polar transport inhibitors from Saussurea costus and Arctium lappa, and Artemisia absinthium, respectively. About 50 and 40 % inhibitions of auxin polar transport in radish hypocotyl segments were observed at 2.5 μg/plant pre-treatment (see “Materials and methods”) of dehydrocostus lactone and 4-hydroxy-β-thujone, respectively. Although the mode of action of these compounds in inhibiting auxin polar transport has not been clear yet, their possible mechanisms are discussed.     

Molecular fate of thidiazuron and its effects on auxin transport in hypocotyls tissues of Pelargonium × hortorum Bailey

Thidiazuron, a synthetic phenylurea-type cytokinin, has previously beenfound to induce somatic embryogenesis and organogenesis in a wide range ofplantspecies and to modulate the metabolism of endogenous auxins and cytokinins. Inspite of these findings, the precise mode of action of TDZ remainsundetermined.The current studies were undertaken to determine the fate of the TDZ moleculeand the effects of TDZ exposure on auxin transport in plants. The fate of tworadiolabelled versions of thidiazuron, [¹⁴C-5-thidiazol]-TDZ and[¹⁴C-U-phenyl]-TDZ, was investigated in sterile hypocotyl culturesofgeranium (Pelargonium×hortorumBailey). Radiolabelled TDZ was recovered from the tissue explants inethanol-insoluble, ethanol-soluble and chloroform fractions as well as inacidic, basic and neutral eluants from Dowex resins. Hypocotyl sections thathadbeen exposed to TDZ were found to accumulate more ¹⁴C-IAA from theculture medium and to translocate the auxin over a greater distance within thetissues. These data provide the first evidence that the TDZ molecule remainsintact in both a free and conjugated form within the plant tissues and providesome indication that TDZ-exposure enhances the accumulation and translocationofauxin within the tissues.     

Polar transport of auxin: carrier-mediated flux across the plasma membrane or neurotransmitter-like secretion?

Auxin (indole-3-acetic acid) has its name derived from the Greek word auxein, meaning 'to increase', and it drives plant growth and development. Auxin is a small molecule derived from the amino acid tryptophan and has both hormone- and morphogen-like properties. Although there is much still to be learned, recent progress has started to unveil how auxin is transported from cell-to-cell in a polar manner. Two recent breakthrough papers from Gerd Jürgens' group indicate that auxin transport is mediated by regulated vesicle trafficking, thus encompassing neurotransmitter-like features.     

The discovery of Arylex™ active and Rinskor™ active: Two novel auxin herbicides

Multiple classes of commercially important auxin herbicides have been discovered since the 1940s including the aryloxyacetates (2,4-D, MCPA, dichlorprop, mecoprop, triclopyr, and fluroxypyr), the benzoates (dicamba), the quinoline-2-carboxylates (quinclorac and quinmerac), the pyrimidine-4-carboxylates (aminocyclopyrachlor), and the pyridine-2-carboxylates (picloram, clopyralid, and aminopyralid). In the last 10 years, two novel pyridine-2-carboxylate (or picolinate) herbicides were discovered at Dow AgroSciences. This paper will describe the structure activity relationship study that led to the discovery of the 6-aryl-picolinate herbicides Arylex™ active (2005) and Rinskor™ active (2010). While Arylex was developed primarily for use in cereal crops and Rinskor is still in development primarily for use in rice crops, both herbicides will also be utilized in additional crops.     

Models of long-distance transport: how is carrier-dependent auxin transport regulated in the stem?

? This paper presents two models of carrier-dependent long-distance auxin transport in stems that represent the process at different scales. ? A simple compartment model using a single constant auxin transfer rate produced similar data to those observed in biological experiments. The effects of different underlying biological assumptions were tested in a more detailed model representing cellular and intracellular processes that enabled discussion of different patterns of carrier-dependent auxin transport and signalling. ? The output that best fits the biological data is produced by a model where polar auxin transport is not limited by the number of transporters/carriers and hence supports biological data showing that stems have considerable excess capacity to transport auxin. ? All results support the conclusion that auxin depletion following apical decapitation in pea (Pisum sativum) occurs too slowly to be the initial cause of bud outgrowth. Consequently, changes in auxin content in the main stem and changes in polar auxin transport/carrier abundance in the main stem are not correlated with axillary bud outgrowth.     

Genetic approach towards the identification of auxin–cytokinin crosstalk components involved in root development

Phytohormones are important plant growth regulators that control many developmental processes, such as cell division, cell differentiation, organogenesis and morphogenesis. They regulate a multitude of apparently unrelated physiological processes, often with overlapping roles, and they mutually modulate their effects. These features imply important synergistic and antagonistic interactions between the various plant hormones. Auxin and cytokinin are central hormones involved in the regulation of plant growth and development, including processes determining root architecture, such as root pole establishment during early embryogenesis, root meristem maintenance and lateral root organogenesis. Thus, to control root development both pathways put special demands on the mechanisms that balance their activities and mediate their interactions. Here, we summarize recent knowledge on the role of auxin and cytokinin in the regulation of root architecture with special focus on lateral root organogenesis, discuss the latest findings on the molecular mechanisms of their interactions, and present forward genetic screen as a tool to identify novel molecular components of the auxin and cytokinin crosstalk.     

Vascular patterning: more than just auxin?

The plant hormone auxin has long been known to play a pivotal role in vascular patterning and differentiation. But auxin is not the whole story: recent genetic analyses have identified additional factors required for vascular patterning, one of them involving sterols.     

Do we have the auxin receptor yet?

Several auxin-binding proteins (ABP) have now been identified using a variety of techniques. A 43-kDa glycoprotein thought to be a dimer of 22-kDa subunits has been identified as a strong candidate for the auxin receptor that mediates cell elongation in etiolated maize shoots. The primary sequence has been deduced and several interesting structural features have been discerned. There is indirect evidence that this 22-kDa ABP has a receptor function, the most compelling being that antibodies directed against the ABP can block an auxin-induced response. There is evidence that changes in auxin-induced growth capacity in shoots correlates with changes in the abundance of the 22-kDa ABP suggesting that in some cases the 22-kDa ABP may be limiting growth. Confirmation of receptor function for one of these newly-identified ABP's should open the way for genetic manipulation of crop growth.