Auxin Action Research Articles

Auxin production by the Klebsiella planticola strain TSKhA-91 and its effect on development of cucumber (Cucumis sativus L.) seeds

Capacity of Klebsiella planticola strain TSJhA-91 for synthesis of indolyl-3-acetic acid (IAA) and other auxins was studied. The qualitative and quantitative composition of these compounds depends on the presence of exogeneous tryptophan and on the nitrogen source. The highest IAA yield was obtained at the stationary phase of growth. Addition of L-tryptophan to the medium resulted in a significant increase (up to 85.5 microg/mL) of auxin biosynthesis, especially in the presence of nitrates. Thin-layer chromatography revealed that the indole-3-acetamide pathway was not active in this strain. The biological activity of auxins was confirmed by plant assay with the kidney bean cuttings; the height of root formation and rdot number increased 16- and 6-fold, respectively. Under conditions of low-temperature stress, protective effect of K. planticola TSKhA-91 on development of cucumbers (Cucumissativus L.) seeds, including stimulation of germi- nation and root formation by its seeds were shown.

Pollinia-borne chemicals that induce early postpollination effects in Dendrobium flowers move rapidly into agar blocks and include ACC and compounds with auxin activity

The early visible effects of pollination in orchids are likely due to pollinia-borne chemicals. In Dendrobium we tested whether such compounds were water soluble and would diffuse in solid-aqueous phase, and determined both 1-aminocyclopropane-1-carboxylic acid (ACC) concentrations and auxin activity. Following pollination, the flower peduncle showed epinastic movement, followed by yellowing of the flower lip, flower senescence and ovary growth. Placing pollinia on agar blocks for 3, 6, 9 or 12h, prior to transferring them to the stigma, increased the time to these early postpollination effects or prevented them. Placing agar blocks that had been used for contact with the pollinia on the stigma also induced the early postpollination effects. The concentrations of ACC, the direct precursor of ethylene, in pollinia was lower the longer the pollinia had been in contact with the agar blocks, whilst the ACC content in the agar blocks increased with the period of contact. The auxin activity of the agar blocks also increased with the time of contact with pollinia. It is concluded that chemicals in the pollinia are responsible for the early visible postpollination effects, and that these (a) rapidly diffuse in aqueous media, and (b) comprise at least ACC and compounds with auxin activity. The idea is discussed that ACC plus auxin is adequate for the production of the early postpollination effects.

Role of chloride ions in the promotion of auxin-induced growth of maize coleoptile segments.

The mechanism of auxin action on ion transport in growing cells has not been determined in detail. In particular, little is known about the role of chloride in the auxin-induced growth of coleoptile cells. Moreover, the data that do exist in the literature are controversial. This study describes experiments that were carried out with maize (Zea mays) coleoptile segments, this being a classical model system for studies of plant cell elongation growth. Growth kinetics or growth and pH changes were recorded in maize coleoptiles using two independent measuring systems. The growth rate of the segments was measured simultaneously with medium pH changes. Membrane potential changes in parenchymal cells of the segments were also determined for chosen variants. The question of whether anion transport is involved in auxin-induced growth of maize coleoptile segments was primarily studied using anion channel blockers [anthracene-9-carboxylic acid (A-9-C) and 4,4'-diisothiocyanatostilbene-2,2'-disulphonic acid (DIDS)]. In addition, experiments in which KCl was replaced by KNO3 were also performed. Both anion channel blockers, added at 0·1 mm, diminished indole-3-acetic acid (IAA)-induced elongation growth by ~30 %. Medium pH changes measured simultaneously with growth indicated that while DIDS stopped IAA-induced proton extrusion, A-9-C diminished it by only 50 %. Addition of A-9-C to medium containing 1 mm KCl did not affect the characteristic kinetics of IAA-induced membrane potential changes, while in the presence of 10 mm KCl the channel blocker stopped IAA-induced membrane hyperpolarization. Replacement of KCl with KNO3 significantly decreased IAA-induced growth and inhibited proton extrusion. In contrast to the KCl concentration, the concentration of KNO3 did not affect the growth-stimulatory effect of IAA. For comparison, the effects of the cation channel blocker tetraethylammonium chloride (TEA-Cl) on IAA-induced growth and proton extrusion were also determined. TEA-Cl, added 1 h before IAA, caused reduction of growth by 49·9 % and inhibition of proton extrusion. These results suggest that Cl(-) plays a role in the IAA-induced growth of maize coleoptile segments. A possible mechanism for Cl(-) uptake during IAA-induced growth is proposed in which uptake of K(+) and Cl(-) ions in concert with IAA-induced plasma membrane H(+)-ATPase activity changes the membrane potential to a value needed for turgor adjustment during the growth of maize coleoptile cells.

EFECTO DE UN BIORREGULADOR A BASE DE AUXINAS SOBRE EL CRECIMIENTO DE PLANTINES DE TOMATE

Production of tomato seedlings in trays has increased in recent years because protecting the root system in a ball reduces transplant shock, resulting in a higher plant survival rate and thus avoiding the need to replant. Greater root development favors establishment in the field because it promotes water and nutrient uptake. Since root production can be promoted by auxin action, the aim of this study was to evaluate the effect of an auxin-based bioregulator on the growth of tomato seedlings in trays. The trial was conducted in a commercial nursery in San Juan, Argentina. The AC 55 tomato cultivar was used. Trays with 126 cells, 36 cm3 per cell, were used. The bioregulator was a commercial formulation containing 0.12 % indole butyric acid and 0.004 % forchlorfenuron. Four concentrations of the product (1, 3, 5 and 7 ml·liter-1) and a control were tested in a completely randomized design with 5 treatments and 4 replicates. Samples were taken at 26, 33, 40 and 47 days after sowing. The variables measured were root dry weight (RDW), aerial dry weight (ADW), stem length (SL), number of leaves larger than 1 cm (NL) and third leaf length (LL). Higher doses increased RDW by 36-39 % compared to the control. The regression between root dry weight and applied dose indicates there could be a response to higher doses. No effect on growth in the aerial part of the plant was observed.

Auxin transport sites are visualized in planta using fluorescent auxin analogs.

The plant hormone auxin is a key morphogenetic signal that controls many aspects of plant growth and development. Cellular auxin levels are coordinately regulated by multiple processes, including auxin biosynthesis and the polar transport and metabolic pathways. The auxin concentration gradient determines plant organ positioning and growth responses to environmental cues. Auxin transport systems play crucial roles in the spatiotemporal regulation of the auxin gradient. This auxin gradient has been analyzed using SCF-type E3 ubiquitin-ligase complex-based auxin biosensors in synthetic auxin-responsive reporter lines. However, the contributions of auxin biosynthesis and metabolism to the auxin gradient have been largely elusive. Additionally, the available information on subcellular auxin localization is still limited. Here we designed fluorescently labeled auxin analogs that remain active for auxin transport but are inactive for auxin signaling and metabolism. Fluorescent auxin analogs enable the selective visualization of the distribution of auxin by the auxin transport system. Together with auxin biosynthesis inhibitors and an auxin biosensor, these analogs indicated a substantial contribution of local auxin biosynthesis to the formation of auxin maxima at the root apex. Moreover, fluorescent auxin analogs mainly localized to the endoplasmic reticulum in cultured cells and roots, implying the presence of a subcellular auxin gradient in the cells. Our work not only provides a useful tool for the plant chemical biology field but also demonstrates a new strategy for imaging the distribution of small-molecule hormones.

A model for an early role of auxin in Arabidopsis gynoecium morphogenesis.

The female reproductive organ of angiosperms, the gynoecium, often consists of the fusion of multiple ovule-bearing carpels. It serves the important function of producing and protecting ovules as well as mediating pollination. The gynoecium has likely contributed to the tremendous success of angiosperms over their 160 million year history. In addition, being a highly complex plant organ, the gynoecium is well suited to serving as a model system for use in the investigation of plant morphogenesis and development. The longstanding model of gynoecium morphogenesis in Arabidopsis holds that apically localized auxin biosynthesis in the gynoecium results in an apical to basal gradient of auxin that serves to specify along its length the development of style, ovary, and gynophore in a concentration-dependent manner. This model is based primarily on the observed effects of the auxin transport blocker N-1-naphthylphthalamic acid (NPA) as well as analyses of mutants of Auxin Response Factor (ARF) 3/ETTIN (ETT). Both NPA treatment and ett mutation disrupt gynoecium morphological patterns along the apical–basal axis. More than a decade after the model’s initial proposal, however, the auxin gradient on which the model critically depends remains elusive. Furthermore, multiple observations are inconsistent with such an auxin-gradient model. Chiefly, the timing of gynoecium emergence and patterning occurs at a very early stage when the organ has little-to-no apical–basal dimension. Based on these observations and current models of early leaf patterning, we propose an alternate model for gynoecial patterning. Under this model, the action of auxin is necessary for the early establishment of adaxial–abaxial patterning of the carpel primordium. In this case, the observed gynoecial phenotypes caused by NPA and ett are due to the disruption of this early adaxial–abaxial patterning of the carpel primordia. Here we present the case for this model based on recent literature and current models of leaf development.

A Long Noncoding Way to Alternative Splicing in Plant Development

In this issue of Developmental Cell, Bardou et al. (2014) elucidate how long, highly structured noncoding RNAs control alternative splicing regulators that specifically mediate the action of the hormone auxin in the promotion of lateral root growth in Arabidopsis.

Auxin sensitivities of all Arabidopsis Aux/IAAs for degradation in the presence of every TIR1/AFB.

Auxin plays a key role in regulation of almost all processes of plant growth and development. Different physiological processes are regulated by different ranges of auxin concentrations; however, the underlying mechanisms creating these differences are largely unknown. The first step of auxin signaling is auxin-dependent interaction of an auxin receptor with transcriptional co-repressors (Aux/IAA), which leads to Aux/IAA degradation. Arabidopsis has six homologous auxin receptors (TIR1 and five AFBs), 29 Aux/IAA proteins and two types of active auxins, IAA and phenylacetic acid (PAA). Therefore, a large number of possible combinations between these three factors may contribute to the creation of complex auxin responses. Using a yeast heterologous reconstitution system, we investigated auxin-dependent degradation of all Arabidopsis Aux/IAAs in combination with every TIR or AFB receptor component. We found that TIR1 and AFB2 were effective in mediating Aux/IAA degradation. We confirmed that the Aux/IAA domain II, which binds TIR1, is essential for degradation. IAA and other natural auxins, 4-chloroindole-3-acetic acid (4-Cl-IAA) and PAA, induced Aux/IAA degradation; and IAA and 4-Cl-IAA had higher activity than PAA. Effective auxin concentrations for Aux/IAA degradation depended on both Aux/IAAs and TIR1 or AFB2 receptors, which is consistent with the Aux/IAA-TIR1/AFB co-receptor concept.

PIN-driven polar auxin transport in plant developmental plasticity: a key target for environmental and endogenous signals.

Plants master the art of coping with environmental challenges in two ways: on the one hand, through their extensive defense systems, and on the other, by their developmental plasticity. The plant hormone auxin plays an important role in a plant's adaptations to its surroundings, as it specifies organ orientation and positioning by regulating cell growth and division in response to internal and external signals. Important in auxin action is the family of PIN-FORMED (PIN) auxin transport proteins that generate auxin maxima and minima by driving polar cell-to-cell transport of auxin through their asymmetric subcellular distribution. Here, we review how regulatory proteins, the cytoskeleton, and membrane trafficking affect PIN expression and localization. Transcriptional regulation of PIN genes alters protein abundance, provides tissue-specific expression, and enables feedback based on auxin concentrations and crosstalk with other hormones. Post-transcriptional modification, for example by PIN phosphorylation or ubiquitination, provides regulation through protein trafficking and degradation, changing the direction and quantity of the auxin flow. Several plant hormones affect PIN abundance, resulting in another means of crosstalk between auxin and these hormones. In conclusion, PIN proteins are instrumental in directing plant developmental responses to environmental and endogenous signals.

A 2,4-dichlorophenoxyacetic acid analog screened using a maize coleoptile system potentially inhibits indole-3-acetic acid influx in Arabidopsis thaliana

Studies using inhibitors of indole-3-acetic acid (IAA) transport, not only for efflux but influx carriers, provide many aspects of auxin physiology in plants. 1-Naphtoxyacetic acid (1-NOA), an analog of the synthetic auxin 1-N-naphtalene acetic acid (NAA), inhibits the IAA influx carrier AUX1. However, 1-NOA also shows auxin activity because of its structural similarity to NAA. In this study, we have identified another candidate inhibitor of the IAA influx carrier. The compound, “7-B3; ethyl 2-[(2-chloro-4-nitrophenyl)thio]acetate,” is a 2,4-dichlorophenoxyacetic acid (2,4-D) analog. At high concentrations (> 300 µM), 7-B3 slightly reduced IAA transport and tropic curvature of maize coleoptiles, whereas lower concentrations had almost no effect. We have analyzed the effects of 7-B3 on Arabidopsis thaliana seedlings. 7-B3 rescued the 2,4-D-inhibited root elongation, but not the NAA-inhibited root elongation. The effect of 7-B3 was weaker than that of 1-NOA. Both 1-NOA and 7-B3 inhibited DR5::GUS expression induced by IAA and 2,4-D, but not that induced by NAA. At high concentrations, 1-NOA exhibited auxin activity, but 7-B3 did not. Furthermore, 7-B3 inhibited apical hook formation in etiolated seedlings more effectively than 1-NOA did. These results indicate that 7-B3 is a potential inhibitor of IAA influx that has almost no effect on IAA efflux or auxin signaling.

Tungsten disrupts root growth in Arabidopsis thaliana by PIN targeting

Tungsten is a heavy metal with increasing concern over its environmental impact. In plants it is extensively used to deplete nitric oxide by inhibiting nitrate reductase, but its presumed toxicity as a heavy metal has been less explored. Accordingly, its effects on Arabidopsis thaliana primary root were assessed. The effects on root growth, mitotic cell percentage, nitric oxide and hydrogen peroxide levels, the cytoskeleton, cell ultrastructure, auxin and cytokinin activity, and auxin carrier distribution were investigated. It was found that tungsten reduced root growth, particularly by inhibiting cell expansion in the elongation zone, so that root hairs emerged closer to the root tip than in the control. Although extensive vacuolation was observed, even in meristematic cells, cell organelles were almost unaffected and microtubules were not depolymerized but reoriented. Tungsten affected auxin and cytokinin activity, as visualized by the DR5-GFP and TCS-GFP expressing lines, respectively. Cytokinin fluctuations were similar to those of the mitotic cell percentage. DR5-GFP signal appeared ectopically expressed, while the signals of PIN2-GFP and PIN3-GFP were diminished even after relatively short exposures. The observed effects were not reminiscent of those of any nitric oxide scavengers. Taken together, inhibition of root growth by tungsten might rather be related to a presumed interference with the basipetal flow of auxin, specifically affecting cell expansion in the elongation zone.

Phytomelatonin: Discovery, Content, and Role in Plants

Melatonin (N-acetyl-5-methoxytryptamine) is an indolic compound derived from tryptophan. Usually identified as a neurotransmitter or animal hormone, this compound was detected in plants in 1995. Interest in knowing the melatonin content of plants and its possible role therein is growing, as indicated by the increasing number of related publications. Melatonin is present in all plant species studied, with large variations in its level depending on the plant organ or tissue. It seems to be more abundant in aromatic plants and in leaves than in seeds. Regarding its physiological function in plants, melatonin shows auxin activity and is an excellent antioxidant, regulating the growth of roots, shoots, and explants, activating seed germination and rhizogenesis (lateral- and adventitious-roots), and delaying induced leaf senescence. Its ability to strengthen plants subjected to abiotic stress such as drought, cold, heat, salinity, chemical pollutants, herbicides, and UV radiation makes melatonin an interesting candidate for use as a natural biostimulating substance for treating field crops.

Auxin is a central player in the hormone cross‐talks that control adventitious rooting

Vegetative propagation of economically important woody, horticultural and agricultural species rely on an efficient adventitious root (AR) formation. The formation of ARs is a complex genetic trait regulated by the interaction of environmental and endogenous factors among which the phytohormone auxin plays an essential role. This article summarizes the current knowledge related to the intricate network through which auxin controls adventitious rooting. How auxin and recently identified auxin-related compounds affect AR formation in different plant species is discussed. Particular attention is addressed to illustrate how auxin has a central role in the hormone cross-talk leading to AR development. In parallel, we describe the molecular players involved in the control of auxin homeostasis, transport and signaling, for a better understanding of the auxin action during adventitious rooting.

Mechanisms regulating auxin action during fruit development

Auxin controls many aspects of fruit development, including fruit set and growth, ripening and abscission. However, the mechanisms by which auxin regulates these processes are still poorly understood. While it is generally agreed that precise spatial and temporal control of auxin distribution and signaling are required for fruit development, the dynamics of auxin biosynthesis and the mechanisms for its transport to different fruit tissues are mostly unknown. Despite major advances in elucidating many aspects of auxin biology in vegetative tissues, until recently, the nature and importance of auxin metabolism, transport and signaling during fruit ontogeny remained obscure. In this review, we summarize recent research that has started to elucidate the molecular mechanisms by which auxin is produced and transported in the fruit and to unravel the complexity of auxin signaling during fruit development. We also discuss recent approaches used to reveal the genes and regulatory networks that mediate cell and tissue-specific control of auxin levels in the developing fruit.

Hormonal control of cell division and elongation along differentiation trajectories in roots

The continuous development of roots is supported by a sustainable system for cell production and growth at the root tip. In the stem cell niche that consists of a quiescent centre and surrounding stem cells, an undifferentiated state and low mitotic activity are preserved by the action of auxin and abscisic acid. Stem cell daughters divide several times in the proximal meristem, where auxin and gibberellin mainly promote cell proliferation. Cells then elongate with the help of gibberellin, and become finally differentiated as a constituent of a cell file in the elongation/differentiation zone. In the model plant Arabidopsis thaliana, the transition zone is located between the proximal meristem and the elongation/differentiation zone, and plays an important role in switching from mitosis to the endoreplication that causes DNA polyploidization. Recent studies have shown that cytokinins are essentially required for this transition by antagonizing auxin signalling and promoting degradation of mitotic regulators. In each root zone, different phytohormones interact with one another and coordinately control cell proliferation, cell elongation, cell differentiation, and endoreplication. Such hormonal networks maintain the elaborate structure of the root tip under various environmental conditions. In this review, we summarize and discuss key issues related to hormonal regulation of root growth, and describe how phytohormones are associated with the control of cell cycle machinery.

Auxin transport and activity regulate stomatal patterning and development

Stomata are two-celled valves that control epidermal pores whose spacing optimizes shoot-atmosphere gas exchange. They develop from protodermal cells after unequal divisions followed by an equal division and differentiation. The concentration of the hormone auxin, a master plant developmental regulator, is tightly controlled in time and space, but its role, if any, in stomatal formation is obscure. Here dynamic changes of auxin activity during stomatal development are monitored using auxin input (DII-VENUS) and output (DR5:VENUS) markers by time-lapse imaging. A decrease in auxin levels in the smaller daughter cell after unequal division presages the acquisition of a guard mother cell fate whose equal division produces the two guard cells. Thus, stomatal patterning requires auxin pathway control of stem cell compartment size, as well as auxin depletion that triggers a developmental switch from unequal to equal division.

Ethylene in the control of photoperiodic flower induction in Pharbitis nil Chois.

The content of endogenous ethylene in the seedlings of <em>Pharbitis nil</em> subjected to 16-hour long inductive night is low during the first half of a dark period, then it increases considerably in the second half of the night. Ethrel, the compound releasing ethylene, applied to the cotyledons of the seedlings, increases the amount of endogenous ethylene in them and at the same time inhibits the flowering, especially when ethrel was applied during the first half of an inductive night, when the content of endogenous ethylene in the seedlings is low. The auxin, inhibiting the flowering of <em>Pharbitis</em>, causes at the same time the increase in the production of endogenous ethylene. PCIB, an inhibitor of auxin action reverses the inhibiting influence of ethrel on flowering. On the other hand the combined application of ethrel and TIBA, the inhibitor of auxin polar transport, causes the increase of the flowering inhibition. CoCl<sub>2</sub>, the inhibitor of ethylene biosynthesis, and AgNO<sub>3</sub>, the inhibitor of ethylene action, reverse partly the inhibiting influence of auxin. It suggests that ethylene could take part in auxininhibition of flowering. The all obtained results seem to suggest the participation of ethylene in the control of the flower photoperiodic induction.

Decrease of Auxin Binding Protein 1 Gene Expression Alters Shoot Development in Ramie

AUXIN BINDING PROTEIN1 (ABP1) has long been proposed as a candidate of auxin receptor to mediate auxin action in plants. It is essential for plant organ development. The function of ABP1 in the development of ramie (Boehmeria nivea L.) remains unclear. Here, we examined the role of the ABP1 gene in the development of shoot in ramie, especially in leaf venation ontogeny, by transforming ABP1 antisense construct into the plants. We showed that ABP1 expression was decreased in ABP1 antisense transgenic plantlets. The decrease of ABP1 expression led to defects in plant growth including dwarf plant, and decreased apical dominance in the intact plant. In addition, ABP1 antisense lines exhibited a serious retard of leaf development involving small and twisted leaves, sparse trichome, and a decreased cell expansion. Moreover, the decreased ABP1 expression repressed the development of leaf venation. ABP1 antisense lines developed an incomplete leaf venation, which became smaller and lacked in some leaf tissues. Though the cell arrangement was normal in midrib and lateral vein, cell size was decrease and some cell layers were missing in midrib. The abnormal phenotype of leaf venation was attributed to the decreases in cell number and size. Our data support that ABP1 is necessary for shoot growth in ramie. More importantly, it plays a key role for the development of ramie leaf venation by regulating both cell expansion and division.

IAA in the control of photoperiodic flower induction of Pharbitis nil chois

The endogenous content of IAA in the cotyledons of <i>Pharbitis nil</i> is low before and during the first half of the inductive 16-h-long dark period. From the 8th to the 12th hour the level of IAA increased and then again was going down at the end of a dark period. Exogenous IAA applied to the cotyledones before and during the first half of the inductive dark period inhibits flower bud formation. The application of IAA to shoot apex also resulted in the inhibition of flowering. Experiments with TIBA, an auxin polar transport inhibitor, and PCIB, an auxin action inhibitor, have shown that auxin polar transport in cotyledones and long-distance auxin transport from cotyledones to shoot apex play an important role in IAA inhibition of flower bud formation. It suggests that auxins play their role not only at the level of floral induction in cotyledones, but also in the later events of floral evocation and differentiation in shoot apex.

Hormonal regulation during plant fruit development

The modern concept of the hormonal regulation of fruit set, growth, maturation, and ripening is considered. Pollination and fertilization induce ovule activation by surmounting the blocking action of ethylene and ABA to be manifested in auxin accumulation. Active fruit growth by pericarp cell division and elongation is due to the syntheses ofauxin in the developing seed and ofgibberellins in the pericarp. In climacteric fleshy fruits, the maturation is controlled by ethylene via so-called System 1 combining the possibilities of autoinhibition and autocatalysis by ethylene of its own biosynthesis. Transition of tomato fruits from maturation to ripening is characterized by highly active synthesis of ethylene and its receptors due to the functioning of regulatory System 2 resulting in the up-regulation of much greater number of ethylene-inducible genes. In peach fruits, the hormonal regulation of ripening includes also an active auxin involvement in the ethylene biosynthesis, which is combined with the ethylene-induced expression of genes encoding both auxin biosynthesis and the response to auxin. Ethylene induces the expression of genes responsible for the fruit softening, its taste, color, and flavor. Nonclimacteric fleshy fruits produce very small amounts ofethylene; its evolution increases only by the very end of ripening and can be described by a reduced System 1. The ripening of nonclimacteric fruits only weakly depends on ethylene but is stimulated by abscisic acid.