Most cited article - PubMed ID 28265985
Analytical Determination of Auxins and Cytokinins
Belowground interactions of plants with other organisms in the rhizosphere rely on extensive small-molecule communication. Chemical signals released from host plant roots ensure the development of beneficial arbuscular mycorrhizal (AM) fungi which in turn modulate host plant growth and stress tolerance. However, parasitic plants have adopted the capacity to sense the same signaling molecules and to trigger their own seed germination in the immediate vicinity of host roots. The contribution of AM fungi and parasitic plants to the regulation of phytohormone levels in host plant roots and root exudates remains largely obscure. Here, we studied the hormonome in the model system comprising tobacco as a host plant, Phelipanche spp. as a holoparasitic plant, and the AM fungus Rhizophagus irregularis. Co-cultivation of tobacco with broomrape and AM fungi alone or in combination led to characteristic changes in the levels of endogenous and exuded abscisic acid, indole-3-acetic acid, cytokinins, salicylic acid, and orobanchol-type strigolactones. The hormonal content in exudates of broomrape-infested mycorrhizal roots resembled that in exudates of infested non-mycorrhizal roots and differed from that observed in exudates of non-infested mycorrhizal roots. Moreover, we observed a significant reduction in AM colonization of infested tobacco plants, pointing to a dominant role of the holoparasite within the tripartite system.
- Keywords
- mycorrhizal fungi, parasitic plants, plant hormones, rhizosphere, root exudates, small-molecule communication, strigolactones,
- MeSH
- Chromatography, Liquid MeSH
- Cytokinins metabolism MeSH
- Heterocyclic Compounds, 3-Ring metabolism MeSH
- Mass Spectrometry MeSH
- Fungi physiology MeSH
- Host-Pathogen Interactions MeSH
- Plant Roots metabolism microbiology MeSH
- Abscisic Acid metabolism MeSH
- Salicylic Acid metabolism MeSH
- Indoleacetic Acids metabolism MeSH
- Lactones metabolism MeSH
- Mycorrhizae physiology MeSH
- Orobanche growth & development microbiology MeSH
- Nicotiana growth & development microbiology MeSH
- Publication type
- Journal Article MeSH
- Names of Substances
- Cytokinins MeSH
- GR24 strigolactone MeSH Browser
- Heterocyclic Compounds, 3-Ring MeSH
- indoleacetic acid MeSH Browser
- Abscisic Acid MeSH
- Salicylic Acid MeSH
- Indoleacetic Acids MeSH
- Lactones MeSH
Plant hormones regulate numerous developmental and physiological processes. Abiotic stresses considerably affect production and distribution of phytohormones as the stress signal triggers. The homeostasis of plant hormones is controlled by their de novo synthesis and catabolism. The aim of this work was to analyse the contents of total and individual groups of endogenous cytokinins (CKs) as well as indole-3-acetic acid (IAA) in AtCKX overexpressing centaury plants grown in vitro on graded NaCl concentrations (0, 50, 100, 150, 200 mM). The levels of endogenous stress hormones including abscisic acid (ABA), salicylic acid (SA) and jasmonic acid (JA) were also detected. The elevated contents of total CKs were found in all analysed centaury shoots. Furthermore, increased amounts of all five CK groups, as well as enhanced total CKs were revealed on graded NaCl concentrations in non-transformed and AtCKX roots. All analysed AtCKX centaury lines exhibited decreased amounts of endogenous IAA in shoots and roots. Consequently, the IAA/bioactive CK forms ratios showed a significant variation in the shoots and roots of all AtCKX lines. In shoots and roots of both non-transformed and AtCKX transgenic centaury plants, salinity was associated with an increase of ABA and JA and a decrease of SA content.
- MeSH
- Centaurium growth & development metabolism MeSH
- Cyclopentanes analysis metabolism MeSH
- Cytokinins analysis metabolism MeSH
- Plant Roots growth & development metabolism MeSH
- Abscisic Acid analysis metabolism MeSH
- Salicylic Acid metabolism MeSH
- Indoleacetic Acids analysis metabolism MeSH
- Oxylipins analysis metabolism MeSH
- Plant Growth Regulators analysis metabolism MeSH
- Salt Stress * MeSH
- In Vitro Techniques MeSH
- Plant Shoots growth & development metabolism MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Cyclopentanes MeSH
- Cytokinins MeSH
- indoleacetic acid MeSH Browser
- jasmonic acid MeSH Browser
- Abscisic Acid MeSH
- Salicylic Acid MeSH
- Indoleacetic Acids MeSH
- Oxylipins MeSH
- Plant Growth Regulators MeSH
Together with auxin transport, auxin metabolism is a key determinant of auxin signaling output by plant cells. Enzymatic machinery involved in auxin metabolism is subject to regulation based on numerous inputs, including the concentration of auxin itself. Therefore, experiments characterizing altered auxin availability and subsequent changes in auxin metabolism could elucidate the function and regulatory role of individual elements in the auxin metabolic machinery. Here, we studied auxin metabolism in auxin-dependent tobacco BY-2 cells. We revealed that the concentration of N-(2-oxindole-3-acetyl)-l-aspartic acid (oxIAA-Asp), the most abundant auxin metabolite produced in the control culture, dramatically decreased in auxin-starved BY-2 cells. Analysis of the transcriptome and proteome in auxin-starved cells uncovered significant downregulation of all tobacco (Nicotiana tabacum) homologs of Arabidopsis (Arabidopsis thaliana) DIOXYGENASE FOR AUXIN OXIDATION 1 (DAO1), at both transcript and protein levels. Auxin metabolism profiling in BY-2 mutants carrying either siRNA-silenced or CRISPR-Cas9-mutated NtDAO1, as well as in dao1-1 Arabidopsis plants, showed not only the expected lower levels of oxIAA, but also significantly lower abundance of oxIAA-Asp. Finally, ability of DAO1 to oxidize IAA-Asp was confirmed by an enzyme assay in AtDAO1-producing bacterial culture. Our results thus represent direct evidence of DAO1 activity on IAA amino acid conjugates.
- MeSH
- Amino Acids metabolism MeSH
- Dioxygenases metabolism MeSH
- Oxidation-Reduction MeSH
- Plant Proteins metabolism MeSH
- Nicotiana enzymology MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Amino Acids MeSH
- Dioxygenases MeSH
- Plant Proteins MeSH
Cytokinins are a class of phytohormones, signalling molecules specific to plants. They act as regulators of diverse physiological processes in complex signalling pathways. It is necessary for plants to continuously regulate cytokinin distribution among different organs, tissues, cells, and compartments. Such regulatory mechanisms include cytokinin biosynthesis, metabolic conversions and degradation, as well as cytokinin membrane transport. In our review, we aim to provide a thorough picture of the latter. We begin by summarizing cytokinin structures and physicochemical properties. Then, we revise the elementary thermodynamic and kinetic aspects of cytokinin membrane transport. Next, we review which membrane-bound carrier proteins and protein families recognize cytokinins as their substrates. Namely, we discuss the families of "equilibrative nucleoside transporters" and "purine permeases", which translocate diverse purine-related compounds, and proteins AtPUP14, AtABCG14, AtAZG1, and AtAZG2, which are specific to cytokinins. We also address long-distance cytokinin transport. Putting all these pieces together, we finally discuss cytokinin distribution as a net result of these processes, diverse in their physicochemical nature but acting together to promote plant fitness.
- Keywords
- ABCG14, AZG1, AZG2, PUP14, cytokinin distribution, cytokinin hydrophobicity, cytokinin transport, membrane transport,
- MeSH
- Arabidopsis metabolism MeSH
- Biological Transport MeSH
- Cell Membrane metabolism MeSH
- Cytokinins metabolism MeSH
- Homeostasis MeSH
- Hydrophobic and Hydrophilic Interactions MeSH
- Kinetics MeSH
- Plant Roots metabolism MeSH
- Membrane Transport Proteins metabolism MeSH
- Arabidopsis Proteins genetics MeSH
- Gene Expression Regulation, Plant MeSH
- Plant Growth Regulators metabolism MeSH
- Signal Transduction physiology MeSH
- Thermodynamics MeSH
- Plant Shoots metabolism MeSH
- Publication type
- Journal Article MeSH
- Review MeSH
- Names of Substances
- Cytokinins MeSH
- Membrane Transport Proteins MeSH
- Arabidopsis Proteins MeSH
- Plant Growth Regulators MeSH
The plant selective autophagy cargo receptor neighbour of breast cancer 1 gene (NBR1) has been scarcely studied in the context of abiotic stress. We wanted to expand this knowledge by using Arabidopsis thaliana lines with constitutive ectopic overexpression of the AtNBR1 gene (OX lines) and the AtNBR1 Knock-Out (KO lines). Transcriptomic analysis of the shoots and roots of one representative OX line indicated differences in gene expression relative to the parental (WT) line. In shoots, many differentially expressed genes, either up- or down-regulated, were involved in responses to stimuli and stress. In roots the most significant difference was observed in a set of downregulated genes that is mainly related to translation and formation of ribonucleoprotein complexes. The link between AtNBR1 overexpression and abscisic acid (ABA) signalling was suggested by an interaction network analysis of these differentially expressed genes. Most hubs of this network were associated with ABA signalling. Although transcriptomic analysis suggested enhancement of ABA responses, ABA levels were unchanged in the OX shoots. Moreover, some of the phenotypes of the OX (delayed germination, increased number of closed stomata) and the KO lines (increased number of lateral root initiation sites) indicate that AtNBR1 is essential for fine-tuning of the ABA signalling pathway. The interaction of AtNBR1 with three regulatory proteins of ABA pathway (ABI3, ABI4 and ABI5) was observed in planta. It suggests that AtNBR1 might play role in maintaining the balance of ABA signalling by controlling their level and/or activity.
- MeSH
- Arabidopsis genetics metabolism MeSH
- Autophagy * MeSH
- Plants, Genetically Modified MeSH
- Germination MeSH
- Abscisic Acid metabolism MeSH
- Arabidopsis Proteins genetics metabolism MeSH
- Gene Expression Regulation, Plant MeSH
- Seeds genetics MeSH
- Seedlings MeSH
- Signal Transduction * MeSH
- Carrier Proteins genetics metabolism MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Abscisic Acid MeSH
- NBR1 protein, Arabidopsis MeSH Browser
- Arabidopsis Proteins MeSH
- Carrier Proteins MeSH
BACKGROUND AND AIMS: We have recently shown that an Arabidopsis thaliana double mutant of type III phosphatidylinositol-4-kinases (PI4Ks), pi4kβ1β2, constitutively accumulated a high level of salicylic acid (SA). By crossing this pi4kβ1β2 double mutant with mutants impaired in SA synthesis (such as sid2 impaired in isochorismate synthase) or transduction, we demonstrated that the high SA level was responsible for the dwarfism phenotype of the double mutant. Here we aimed to distinguish between the SA-dependent and SA-independent effects triggered by the deficiency in PI4Kβ1 and PI4Kβ2. METHODS: To achieve this we used the sid2pi4kβ1β2 triple mutant. High-throughput analyses of phytohormones were performed on this mutant together with pi4kβ1β2 and sid2 mutants and wild-type plants. Responses to pathogens, namely Hyaloperonospora arabidopsidis, Pseudomonas syringae and Botrytis cinerea, and also to the non-host fungus Blumeria graminis, were also determined. Callose accumulation was monitored in response to flagellin. KEY RESULTS: We show here the prominent role of high SA levels in influencing the concentration of many other tested phytohormones, including abscisic acid and its derivatives, the aspartate-conjugated form of indole-3-acetic acid and some cytokinins such as cis-zeatin. We show that the increased resistance of pi4kβ1β2 plants to the host pathogens H. arabidopsidis, P. syringae pv. tomato DC3000 and Bothrytis cinerea is dependent on accumulation of high SA levels. In contrast, accumulation of callose in pi4kβ1β2 after flagellin treatment was independent of SA. Concerning the response to Blumeria graminis, both callose accumulation and fungal penetration were enhanced in the pi4kβ1β2 double mutant compared to wild-type plants. Both of these processes occurred in an SA-independent manner. CONCLUSIONS: Our data extensively illustrate the influence of SA on other phytohormone levels. The sid2pi4kβ1β2 triple mutant revealed the role of PI4Kβ1/β2 per se, thus showing the importance of these enzymes in plant defence responses.
- Keywords
- Arabidopsis thaliana, biotic stress, callose, isochorismate synthase 1, pathogens, phytohormones, pi4kβ1β2/PI4Ks, salicylic acid,
- MeSH
- 1-Phosphatidylinositol 4-Kinase * MeSH
- Arabidopsis * MeSH
- Salicylic Acid MeSH
- Mutation MeSH
- Plant Diseases MeSH
- Arabidopsis Proteins genetics MeSH
- Pseudomonas syringae MeSH
- Gene Expression Regulation, Plant MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- 1-Phosphatidylinositol 4-Kinase * MeSH
- AT5G09350 protein, Arabidopsis MeSH Browser
- AT5G64070 protein, Arabidopsis MeSH Browser
- Salicylic Acid MeSH
- Arabidopsis Proteins MeSH
The integrity of the actin cytoskeleton is essential for plant immune signalling. Consequently, it is generally assumed that actin disruption reduces plant resistance to pathogen attack. Here, we demonstrate that actin depolymerization induced a dramatic increase in salicylic acid (SA) levels in Arabidopsis thaliana. Transcriptomic analysis showed that the SA pathway was activated due to the action of isochorismate synthase (ICS). The effect was also confirmed in Brassica napus. This raises the question of whether actin depolymerization could, under particular conditions, lead to increased resistance to pathogens. Thus, we explored the effect of pretreatment with actin-depolymerizing drugs on the resistance of Arabidopsis thaliana to the bacterial pathogen Pseudomonas syringae, and on the resistance of an important crop Brassica napus to its natural fungal pathogen Leptosphaeria maculans. In both pathosystems, actin depolymerization activated the SA pathway, leading to increased plant resistance. To our best knowledge, we herein provide the first direct evidence that disruption of the actin cytoskeleton can actually lead to increased plant resistance to pathogens, and that SA is crucial to this process.
- MeSH
- Actins metabolism MeSH
- Arabidopsis metabolism microbiology MeSH
- Ascomycota pathogenicity MeSH
- Brassica napus metabolism microbiology MeSH
- Intramolecular Transferases metabolism MeSH
- Salicylic Acid metabolism MeSH
- Plant Diseases microbiology MeSH
- Arabidopsis Proteins metabolism MeSH
- Pseudomonas syringae pathogenicity MeSH
- Gene Expression Regulation, Plant physiology MeSH
- Signal Transduction physiology MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Actins MeSH
- Intramolecular Transferases MeSH
- isochorismate synthase MeSH Browser
- Salicylic Acid MeSH
- Arabidopsis Proteins MeSH