Indole-3-acetic Acid Signaling Research Articles

GA4 and IAA were involved in the morphogenesis and development of flowers in Agapanthus praecox ssp. orientalis

The transition from vegetative to reproductive growth represents a major phase change in angiosperms. Hormones play important roles in this process. In this study, gibberellic acid (GA), cytokinins (CKs), indoleacetic acid (IAA), and abscisic acid (ABA) were analyzed during the flowering in Agapanthus praecox ssp. orientalis. Eleven types of endogenous gibberellins in addition to GA1 were detected in various organs. GA9 was detected with the highest concentrations, followed by GA5, GA8, and GA19. However, GA4 was the main bioactive GA that was involved in the regulation of flowering. Eight types of endogenous cytokinins were detected in A. praecox ssp. orientalis, and zeatin, zeatin riboside, zeatin-O-glucoside, and N(6)-isopentenyladenosine-5-monophosphate were present at higher levels throughout the study, of which zeatin plays an important role in the development of various organs. IAA increased by 581% in the shoot tips from the vegetative to inflorescence bud stages and had the most significant changes during flowering. Phytohormone immunolocalization analysis suggested that IAA involved in differentiation and development of each floral organs, GA and zeatin play important roles in floret primordia differentiation and ovule development. Using exogenous plant growth regulators proved that GA signaling regulate the scape elongation and stimulate early-flowering, and IAA signaling is involved in the pedicel and corolla elongation and delay flowering slightly.

NO and IAA Key Regulators in the Shoot Growth Promoting Action of Humic Acid in Cucumis sativus L.

Previous studies have reported that a purified sedimentary humic acid (PHA) was able to increase the concentration of nitric oxide (NO), indole-acetic acid (IAA) and ethylene in cucumber roots. Here, we investigated if these effects are functionally related to the ability of PHA to improve shoot growth. The effect of specific inhibitors of NO, IAA and ethylene functionality and signaling on PHA-induced shoot growth was studied. Likewise, the effect of these inhibitors on the synthesis and activity of the phytoregulators concerned by PHA action in cucumber roots was also explored. The results show that shoot growth promoted by PHA is due to an increase of IAA concentration in the root through both a NO-dependent and a NO-independent pathway. In addition, the increased ethylene production in the root is regulated by an IAA-dependent pathway. Finally, results also showed that the increase of ABA concentration in the root is regulated through both IAA- and ethylene-dependent pathways. In summary, the shoot growth promoting action of PHA involves a complex hormonal network. On one hand, the PHA action is functionally linked to increments in NO and IAA concentration in roots. And on the other hand, PHA action also increases ethylene and ABA root concentration mediated by NO-IAA dependent pathways.

Characterization of the Complex Regulation of AtALMT1 Expression in Response to Phytohormones and Other Inducers

In Arabidopsis (Arabidopsis thaliana), malate released into the rhizosphere has various roles, such as detoxifying rhizotoxic aluminum (Al) and recruiting beneficial rhizobacteria that induce plant immunity. ALUMINUM-ACTIVATED MALATE TRANSPORTER1 (AtALMT1) is a critical gene in these responses, but its regulatory mechanisms remain unclear. To explore the mechanism of the multiple responses of AtALMT1, we profiled its expression patterns in wild-type plants, in transgenic plants harboring various deleted promoter constructs, and in mutant plants with defects in signal transduction in response to various inducers. AtALMT1 transcription was clearly induced by indole-3-acetic acid (IAA), abscisic acid (ABA), low pH, and hydrogen peroxide, indicating that it was able to respond to multiple signals, while it was not induced by methyl jasmonate and salicylic acid. The IAA-signaling double mutant nonphototropic hypocotyls4-1; auxin-responsive factor19-1 and the ABA-signaling mutant aba insensitive1-1 did not respond to auxin and ABA, respectively, but both showed an Al response comparable to that of the wild type. A synthetic microbe-associated molecular pattern peptide, flagellin22 (flg22), induced AtALMT1 transcription but did not induce the transcription of IAA- and ABA-responsive biomarker genes, indicating that both Al and flg22 responses of AtALMT1 were independent of IAA and ABA signaling. An in planta β-glucuronidase reporter assay identified that the ABA response was regulated by a region upstream (-317 bp) from the first ATG codon, but other stress responses may share critical regulatory element(s) located between -292 and -317 bp. These results illustrate the complex regulation of AtALMT1 expression during the adaptation to abiotic and biotic stresses.

The Manipulation of Auxin in the Abscission Zone Cells of Arabidopsis Flowers Reveals That Indoleacetic Acid Signaling Is a Prerequisite for Organ Shedding

A number of novel strategies were employed to examine the role of indoleacetic acid (IAA) in regulating floral organ abscission in Arabidopsis (Arabidopsis thaliana). Analysis of auxin influx facilitator expression in β-glucuronidase reporter plants revealed that AUXIN RESISTANT1, LIKE AUX1, and LAX3 were specifically up-regulated at the site of floral organ shedding. Flowers from mutants where individual family members were down-regulated exhibited a reduction in the force necessary to bring about petal separation; however, the effect was not additive in double or quadruple mutants. Using the promoter of a polygalacturonase (At2g41850), active primarily in cells undergoing separation, to drive expression of the bacterial genes iaaL and iaaM, we have shown that it is possible to manipulate auxin activity specifically within the floral organ abscission zone (AZ). Analysis of petal breakstrength reveals that if IAA AZ levels are reduced, shedding takes place prematurely, while if they are enhanced, organ loss is delayed. The At2g41850 promoter was also used to transactivate the gain-of-function AXR3-1 gene in order to disrupt auxin signaling specifically within the floral organ AZ cells. Flowers from transactivated lines failed to shed their sepals, petals, and anthers during pod expansion and maturity, and these organs frequently remained attached to the plant even after silique desiccation and dehiscence had taken place. These observations support a key role for IAA in the regulation of abscission in planta and reveal, to our knowledge for the first time, a requirement for a functional IAA signaling pathway in AZ cells for organ shedding to take place.

Endogenous auxin and jasmonic acid levels are differentially modulated by abiotic stresses in rice

Abiotic stresses such as drought, salinity, and adverse temperatures are major limiting factors for plant growth and reproduction. Plant responses to these stresses are coordinated by arrays of regulatory networks including the induction of endogenous abscisic acid (ABA), a well documented phytohormone for stress responses. However, whether or how these abiotic stresses affect the endogenous biosynthesis or metabolism of other phytohormones remains largely unknown. Here, we report the changes of endogenous indole-3-acetic acid (IAA) and jasmonic acid (JA) levels and expression of genes related to the biosynthesis or signaling of these hormones in rice under various abiotic stress conditions. The IAA content was decreased after drought stress, but it was significantly increased under cold and heat stresses. And the auxin-regulated gravitropism of root tip was inhibited by cold stress. Many genes involved in the IAA biosynthesis and signaling were changed in transcript level under these stresses, and the changes were essentially in agreement with the change of endogenous IAA level. Interestingly, the endogenous JA content was increased markedly under drought and cold stresses, but it was reduced by heat stress. Accordingly, many genes involved in JA biosynthesis and signaling were induced by drought and cold treatment but these genes were significantly suppressed by heat stress. We concluded that endogenous levels of IAA and JA were differentially regulated by abiotic stresses in rice, implying diverse roles of these hormones in stress responses.

Indole-3-acetic acid accumulation during poplar rhizogenesis revealed by immunohistochemistry

Poplar hybrid 741 [Populus alba × (P. davidiana + P. simonii) × P. tomentosa] leaves were rooted within 8 d when cultured in vitro on 1/2 Murashige and Skoog (MS) medium. The spatial distribution of endogenous indole-3-acetic acid (IAA) in the rhizogenesis was investigated, using an immunohistochemical approach. In addition, the effect of 2,3,5-triiodobenzoic acid (TIBA) on IAA distribution was also analyzed. The results showed that a strong IAA signal was detected in the vascular bundles of the basal regions of the petioles 3 d after root induction. Furthermore, the signal in vascular bundles of the basal regions of the petioles was stronger than that of the middle regions of the petioles. Application of TIBA on lamina delayed both the accumulation of IAA in the vascular bundles and rhizogenesis. These data indicate that an endogenous IAA rise in vascular bundles is among the first signals leading to the rhizogenesis, and that it results from transportation of the hormone from the lamina of the leaf to the base of the petiole, rather than by in situ IAA generation.

Immunohistochemical observation of indole-3-acetic acid at the IAA synthetic maize coleoptile tips

To investigate the distribution of IAA (indole-3-acetic acid) and the IAA synthetic cells in maize coleoptiles, we established immunohistochemistry of IAA using an anti-IAA-C-monoclonal antibody. We first confirmed the specificity of the antibody by comparing the amounts of endogenous free and conjugated IAA to the IAA signal obtained from the IAA antibody. Depletion of endogenous IAA showed a corresponding decrease in immuno-signal intensity and negligible cross-reactivity against IAA-related compounds, including tryptophan, indole-3-acetamide, and conjugated-IAA was observed. Immunolocalization showed that the IAA signal was intense in the approximately 1 mm region and the outer epidermis at the approximately 0.5 mm region from the top of coleoptiles treated with 1-N-naphthylphthalamic acid. By contrast, the IAA immuno-signal in the outer epidermis almost disappeared after 5-methyl-tryptophan treatment. Immunogold labeling of IAA with an anti-IAA-N-polyclonal antibody in the outer-epidermal cells showed cytoplasmic localization of free-IAA, but none in cell walls or vacuoles. These findings indicated that IAA is synthesized in the 0–2.0 mm region of maize coleoptile tips from Trp, in which the outer-epidermal cells of the 0.5 mm tip are the most active IAA synthetic cells.

Immunohistochemical Localization of Indole-3-Acetic Acid During Induction of Adventitious Root Formation from Cotyledon Explants of Walnut

Cotyledon explants of walnut (Juglans regia) have been shown to generate adventitious roots on growth regulator-free medium. The spatial distribution of endogenous indole-3-acetic acid (IAA) and its dynamic changes during adventitious root formation were investigated using an in situ immunohistochemical approach. Before root induction, IAA signal was distributed throughout the freshly excised cotyledon explants. During provascular bundle differentiation, the IAA signal was mainly located in the provascular bundles. At the stage of annular meristematic zones formation, the IAA signal was mainly distributed in the meristematic zones and decreased in the vascular bundles and cotyledonous parenchyma. As primordia formed, the IAA signal became localized in the root primordia and gradually disappeared in the meristematic zones. In emerging roots, the IAA signal was mainly localized in the root cap and root meristem. These results suggest that accumulation of IAA in the provascular bundles may induce vascular differentiation and the increase in IAA through meristematic zones may be responsible for the adventitious root formation from walnut cotyledons. The direct evidence presented here indicates that IAA accumulated in the meristematic zones is not the sole signal needed to induce adventitious root.

Approaching Cellular and Molecular Resolution of Auxin Biosynthesis and Metabolism

There is abundant evidence of multiple biosynthesis pathways for the major naturally occurring auxin in plants, indole-3-acetic acid (IAA), and examples of differential use of two general routes of IAA synthesis, namely Trp-dependent and Trp-independent. Although none of these pathways has been completely defined, we now have examples of specific IAA biosynthetic pathways playing a role in developmental processes by way of localized IAA synthesis, causing us to rethink the interactions between IAA synthesis, transport, and signaling. Recent work also points to some IAA biosynthesis pathways being specific to families within the plant kingdom, whereas others appear to be more ubiquitous. An important advance within the past 5 years is our ability to monitor IAA biosynthesis and metabolism at increasingly higher resolution.

Crosstalk between ABA and auxin signaling pathways in roots of Arabidopsis thaliana (L.) Heynh.

Studies of abscisic acid (ABA) and auxin have revealed that these pathways impinge on each other. The Daucus carota (L.) Dc3 promoter: uidA (beta-glucuronidase: GUS) chimaeric reporter (ProDc3:GUS) is induced by ABA, osmoticum, and the auxin indole-3-acetic acid (IAA) in vegetative tissues of transgenic Arabidopsis thaliana (L.) Heynh. Here, we describe the root tissue-specific expression of ProDc3:GUS in the ABA-insensitive-2 (abi2-1), auxin-insensitive-1 (aux1), auxin-resistant-4 (axr4), and rooty (rty1) mutants of Arabidopsis in response to ABA, IAA and synthetic auxins naphthalene acetic acid (NAA), and 2, 4-(dichlorophenoxy) acetic acid. Quantitative analysis of ProDc3:GUS expression showed that the abi2-1 mutant had reduced GUS activity in response to ABA, IAA, or 2, 4-D: , but not to NAA. Similarly, chromogenic staining of ProDc3:GUS activity showed that the aux1 and axr4 mutants gave predictable hypomorphic ProDc3:GUS expression phenotypes in roots treated with IAA or 2, 4-D: , but not the diffusible auxin NAA. Likewise the rty mutant, which accumulates auxin, showed elevated ProDc3:GUS expression in the absence or presence of hormones relative to wild type. Interestingly, the aux1 and axr4 mutants showed a hypomorphic effect on ABA-inducible ProDc3:GUS expression, demonstrating that ABA and IAA signaling pathways interact in roots. Possible mechanisms of crosstalk between ABA and auxin signaling are discussed.

Patterns of auxin distribution during gravitational induction of reaction wood in poplar and pine.

Gravistimulation of tree stems affects wood development by unilaterally inducing wood with modified properties, called reaction wood. Commonly, it also stimulates cambial growth on the reaction wood side. Numerous experiments involving applications of indole-3-acetic acid (IAA) or IAA-transport inhibitors have suggested that reaction wood is induced by a redistribution of IAA around the stem. However, in planta proof for this model is lacking. Therefore, we have mapped endogenous IAA distribution across the cambial region tissues in both aspen (Populus tremula, denoted poplar) and Scots pine (Pinus sylvestris) trees forming reaction wood, using tangential cryosectioning combined with sensitive gas chromatography-mass spectrometry analysis. Moreover, we have documented the kinetics of IAA during reaction wood induction in these species. Our analysis of endogenous IAA demonstrates that reaction wood is formed without any obvious alterations in IAA balance. This is in contrast to gravitropic responses in roots and shoots where a redistribution of IAA has been documented. It is also of interest that cambial growth on the tension wood side was stimulated without an increase in IAA. Taken together, our results suggest a role for signals other than IAA in the reaction wood response, or that the gravitational stimulus interacts with the IAA signal transduction pathway.

Immunohistochemical localization of IAA in developing strawberry fruit

SummaryThe immunohistochemical localization of indole-3-acetic acid (IAA) was carried out by using an anti-indole-3-acetic acid (IAA) monoclonal antibody in the achenes and the receptacles of developing field-grown strawberries in May. The distribution of free-IAA in the achenes and the receptacles and their dynamic variations were established.A small IAA signal was detected in the achenes and the receptacles of strawberry at anthesis. In the receptacles, the IAA signal gradually increased at the immature green (IG) stage. From day 7 after bloom to the inception (IC) stage, the IAA signal was localized throughout the vascular bundles of the receptacle tissues, and the signal strengthened at the early phase of the mature green (MG) stage and at the inception (IC) stage. However, at the firm ripe (FR) stage, the signal was hardly detected in the receptacles. In the achenes, the IAA signal gradually increased from the late phase of the immature green (IG) stage to the late phase of the mature green (MG) stage and was mainly localized in developing embryos. Little signal could be detected after inception (IC) stage in the achenes. The results showed that IAA was especially localized in the phloem of the receptacles and was mainly localized in developing embryos in the achenes, which gave apparent evidence for IAA distribution in fleshy fruits. Moreover, in our research, there was still a strong IAA signal in the receptacles of strawberry at the IC stage, which differed significantly from data in some other reports.

Roles of the reactive oxygen species-generating peroxidase reactions in plant defense and growth induction.

Extracellularly secreted plant peroxidases (POXs) are considered to catalyze the generation of reactive oxygen species (ROS) coupled to oxidation of plant hormone indole-3-acetic acid (IAA) and defense-related compounds salicylic acid (SA), aromatic monoamines (AMAs) and chitooligosaccharides (COSs). This review article consists of two parts, which describe H(2)O(2)-dependent and H(2)O(2)-independent mechanisms for ROS generation, respectively. Recent studies have shown that plant POXs oxidize SA, AMAs and COSs in the presence of H(2)O(2) via a conventional POX cycle, yielding the corresponding radical species, such as SA free radicals. These radical species may react with oxygen, and superoxide (O(2)(.-)) is produced. Through the series of reactions 2 moles of O(2)(.-) can be formed from 1 moles of H(2)O(2), thus leading to oxidative burst. It has been revealed that the ROS induced by SA, AMAs and COSs triggers the increase in cytosolic Ca(2+) concentration. Actually POXs transduce the extracellular signals into the redox signals that eventually stimulate the intracellular Ca(2+) signaling required for induction of defense responses. On the other hand, IAA can react with oxygen and plant POXs in the absence of H(2)O(2), by forming the ternary complex enzyme-IAA-O(2), which readily dissociates into enzyme, IAA radicals and O(2)(.-). This article covers the recent reports showing that extracellularly produced hydroxy radicals derived from O(2)(.-) mediate the IAA-induced cell elongation. Here a novel model for IAA signaling pathway mediated by extracellular ROS produced by cell-wall POXs is proposed. In addition, possible controls of the IAA-POX reactions by a fungal alkaloid are discussed.

Hypaphorine from the ectomycorrhizal fungus Pisolithus tinctorius counteracts activities of indole-3-acetic acid and ethylene but not synthetic auxins in eucalypt seedlings.

Very little is known about the molecules regulating the interaction between plants and ectomycorrhizal fungi during root colonization. The role of fungal auxin in ectomycorrhiza has repeatedly been suggested and questioned, suggesting that, if fungal auxin controls some steps of colonized root development, its activity might be tightly controlled in time and in space by plant and/or fungal regulatory mechanisms. We demonstrate that fungal hypaphorine, the betaine of tryptophan, counteracts the activity of indole-3-acetic acid (IAA) on eucalypt tap root elongation but does not affect the activity of the IAA analogs 2,4-D ((2,4-dichlorophenoxy)acetic acid) or NAA (1-naphthaleneacetic acid). These data suggest that IAA and hypaphorine interact during the very early steps of the IAA perception or signal transduction pathway. Furthermore, while seedling treatment with 1-amincocyclopropane-1-carboxylic acid (ACC), the precursor of ethylene, results in formation of a hypocotyl apical hook, hypaphorine application as well as root colonization by Pisolithus tinctorius, a hypaphorine-accumulating ectomycorrhizal fungus, stimulated hook opening. Hypaphorine counteraction with ACC is likely a consequence of hypaphorine interaction with IAA. In most plant-microbe interactions studied, the interactions result in increased auxin synthesis or auxin accumulation in plant tissues. The P. tinctorius / eucalypt interaction is intriguing because in this interaction the microbe down-regulates the auxin activity in the host plant. Hypaphorine might be the first specific IAA antagonist identified.

Auxin redistributes upwards in graviresponding gynophores of the peanut plant.

The peanut (Arachis hypogaea L.) produces flowers aerially, but buries the recently fertilized ovules into the soil, where fruit and seed development occur. The young seeds are carried down into the soil at the tip of a specialized organ called the gynophore. Although the gynophore has a typical shoot anatomy, it responds positively to gravity like a root. In this study, we explore the role of the plant growth regulator indole-3-acetic acid (IAA) in the growth and the gravitropic response of the peanut gynophore. With an immunolocalization technique using an IAA monoclonal antibody, we localized IAA within the tissues of vertically oriented and gravistimulated gynophores. We found that in vertically oriented gynophores, IAA labeling occurs in the periphery of the gynophore, in the entire cortex and epidermis. Within 20 min of horizontal reorientation, the IAA signal gradually increases in the upper cortex/ epidermis and diminishes in the lower cortex/epidermis. At 1.5 h after gravistimulation, all of the IAA immunolocalization signal is detected in the upper cortex and epidermis--none is detected in the lower side. Growth rate measurements also indicate that after 1-2 h of reorientation, the growth rate maximum on the upper side corresponds temporally and spatially to the growth rate minimum on the lower side. Experiments using radioactively labeled IAA corroborate an upper-side redistribution of this hormone upon horizontal reorientation. These results are analyzed with respect to the current theories of plant gravitropic response, and a model for a possible gravity-induced IAA redistribution from the lower to the upper side of the peanut gynophore is proposed.