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1.
J Biotechnol ; 342: 36-44, 2021 Dec 10.
Article in English | MEDLINE | ID: mdl-34610365

ABSTRACT

Polycyclic aromatic hydrocarbon (PAH) contamination has a negative impact on ecosystems. PAHs are a large group of toxins with two or more benzene rings that are persistent in the environment. Some PAHs can be cytotoxic, teratogenic, and/or carcinogenic. In the bacterium Pseudomonas, PAHs can be modified by dioxygenases, which increase the reactivity of PAHs. We hypothesize that some plant dioxygenases are capable of PAH biodegradation. Herein, we investigate the involvement of Arabidopsis thaliana At1g14130 in the degradation of phenanthrene, our model PAH. The At1g14130 gene encodes Dioxygenase For Auxin Oxidation 1 (AtDAO1), an enzyme involved in the oxidative inactivation of the hormone auxin. Expression analysis using a ß-glucuronidase (GUS) reporter revealed that At1g14130 is prominently expressed in new leaves of plants exposed to media with phenanthrene. Analysis of the oxidative state of gain-of-function mutants showed elevated levels of H2O2 after phenanthrene treatments, probably due to an increase in the oxidation of phenanthrene by AtDAO1. Biochemical assays with purified AtDAO1 and phenanthrene suggest an enzymatic activity towards the PAH. Thus, data presented in this study support the hypothesis that an auxin dioxygenase, AtDAO1, from Arabidopsis thaliana contributes to the degradation of phenanthrene and that there is possible toxic metabolite accumulation after PAH exposure.


Subject(s)
Arabidopsis , Dioxygenases , Phenanthrenes , Polycyclic Aromatic Hydrocarbons , Arabidopsis/genetics , Biodegradation, Environmental , Dioxygenases/genetics , Ecosystem , Hydrogen Peroxide , Indoleacetic Acids , Phenanthrenes/toxicity , Polycyclic Aromatic Hydrocarbons/toxicity
2.
Biochim Biophys Acta Gene Regul Mech ; 1864(3): 194689, 2021 03.
Article in English | MEDLINE | ID: mdl-33561560

ABSTRACT

The extensive processing and protein-assisted stabilization of transcripts have been taken as evidence for a viewpoint that the control of gene expression had shifted entirely in evolution from transcriptional in the bacterial endosymbiont to posttranscriptional in the plastid. This suggestion is however at odds with many observations on plastid gene transcription. Chloroplasts of flowering plants and mosses contain two or more RNA polymerases with distinct promoter preference and division of labor for the coordinated synthesis of plastid RNAs. Plant and algal plastids further possess multiple nonredundant sigma factors that function as transcription initiation factors. The controlled accumulation of plastid sigma factors and modification of their activity by sigma-binding proteins and phosphorylation constitute additional transcriptional regulatory strategies. Plant and algal plastids also contain dedicated one- or two-component transcriptional regulators. Transcription initiation thus continues to form a critical control point at which varied developmental and environmental signals intersect with plastid gene expression.


Subject(s)
Gene Expression Regulation, Plant/physiology , Plastids/metabolism , Transcription Initiation, Genetic/physiology , DNA-Directed RNA Polymerases/genetics , DNA-Directed RNA Polymerases/metabolism , Plant Proteins/genetics , Plant Proteins/metabolism , Plastids/genetics
3.
Photosynth Res ; 147(3): 317-328, 2021 Mar.
Article in English | MEDLINE | ID: mdl-33387192

ABSTRACT

Diatoms are a diverse group of photosynthetic unicellular algae with a plastid of red-algal origin. As prolific primary producers in the ocean, diatoms fix as much carbon as all rainforests combined. The molecular mechanisms that contribute to the high photosynthetic productivity and ecological success of diatoms are however not yet fully understood. Using the model diatom Phaeodactylum tricornutum, here we show rhythmic transcript accumulation of plastid psaA, psbA, petB, and atpB genes as driven by a free running circadian clock. Treatment with the electron transport inhibitor 3-(3,4-dichlorophenyl)-1,1-dimethylurea overrides the circadian signal by markedly downregulating transcription of psaA, petB, and atpB genes but not the psbA gene. Changes in light quantity produce little change in plastid gene transcription while the effect of light quality seems modest with only the psaA gene responding in a pattern that is dependent on the redox state of the plastoquinone pool. The significance of these plastid transcriptional responses and the identity of the underlying genetic control systems are discussed with relevance to diatom photosynthetic acclimation.


Subject(s)
Circadian Rhythm/physiology , Diatoms/metabolism , Gene Expression Regulation/radiation effects , Light , Plastids , Transcription, Genetic/radiation effects , Diatoms/genetics , Humans , Oxidation-Reduction , RNA/physiology , Temperature
4.
Commun Biol ; 3(1): 13, 2020 01 08.
Article in English | MEDLINE | ID: mdl-31925322

ABSTRACT

Photosynthetic efficiency depends on equal light energy conversion by two spectrally distinct, serially-connected photosystems. The redox state of the plastoquinone pool, located between the two photosystems, is a key regulatory signal that initiates acclimatory changes in the relative abundance of photosystems. The Chloroplast Sensor Kinase (CSK) links the plastoquinone redox signal with photosystem gene expression but the mechanism by which it monitors the plastoquinone redox state is unclear. Here we show that the purified Arabidopsis and Phaeodactylum CSK and the cyanobacterial CSK homologue, Histidine kinase 2 (Hik2), are iron-sulfur proteins. The Fe-S cluster of CSK is further revealed to be a high potential redox-responsive [3Fe-4S] center. CSK responds to redox agents with reduced plastoquinone suppressing its autokinase activity. Redox changes within the CSK iron-sulfur cluster translate into conformational changes in the protein fold. These results provide key insights into redox signal perception and propagation by the CSK-based chloroplast two-component system.


Subject(s)
Chloroplasts/metabolism , Histidine Kinase/metabolism , Iron/metabolism , Oxidation-Reduction , Sulfur/metabolism , Arabidopsis Proteins/chemistry , Arabidopsis Proteins/metabolism , Enzyme Activation , Histidine Kinase/chemistry , Iron/chemistry , Photosynthesis , Protein Conformation , Recombinant Proteins , Spectrum Analysis , Structure-Activity Relationship , Sulfur/chemistry
5.
J Hazard Mater ; 321: 268-280, 2017 Jan 05.
Article in English | MEDLINE | ID: mdl-27637093

ABSTRACT

Polycyclic aromatic hydrocarbons (PAHs) are environmental contaminants with cytotoxic, teratogenic and carcinogenic properties. Bioremediation studies with bacteria have led to the identification of dioxygenases (DOXs) in the first step to degrade these recalcitrant compounds. In this study, we characterized the role of the Arabidopsis thaliana AT5G05600, a putative DOX of the flavonol synthase family, in the transformation of PAHs. Phenotypic analysis of loss-of-function mutant lines showed that these plant lines were less sensitive to the toxic effects of phenanthrene, suggesting possible roles of this gene in PAH degradation in vivo. Interestingly, these mutant lines showed less accumulation of H2O2 after PAH exposure. Transgenic lines over-expressing At5g05600 showed a hypersensitive response and more oxidative stress after phenanthrene treatments. Moreover, fluorescence spectra results of biochemical assays with the recombinant His-tagged protein AT5G05600 detected chemical modifications of phenanthrene. Taken together, these results support the hypothesis that AT5G05600 is involved in the catabolism of PAHs and the accumulation of toxic intermediates during PAH biotransformation in plants. This research represents the first step in the design of transgenic plants with the potential to degrade PAHs, leading to the development of vigorous plant varieties that can reduce the levels of these pollutants in the environment.


Subject(s)
Arabidopsis/enzymology , Oxidoreductases/genetics , Plant Proteins/genetics , Plants, Genetically Modified , Polycyclic Aromatic Hydrocarbons/analysis , Soil Pollutants/analysis , Arabidopsis/drug effects , Arabidopsis/genetics , Biodegradation, Environmental , Hydrogen Peroxide , Mutation , Phenanthrenes/analysis , Phenanthrenes/toxicity , Polycyclic Aromatic Hydrocarbons/toxicity , Soil Pollutants/toxicity
6.
Plant Signal Behav ; 10(7): e1027850, 2015.
Article in English | MEDLINE | ID: mdl-26179363

ABSTRACT

Anthocyanins are induced in plants in response to abiotic stresses such as drought, high salinity, excess light, and cold, where they often correlate with enhanced stress tolerance. Numerous roles have been proposed for anthocyanins induced during abiotic stresses including functioning as ROS scavengers, photoprotectants, and stress signals. We have recently found different profiles of anthocyanins in Arabidopsis (Arabidopsis thaliana) plants exposed to different abiotic stresses, suggesting that not all anthocyanins have the same function. Here, we discuss these findings in the context of other studies and show that anthocyanins induced in Arabidopsis in response to various abiotic stresses have different localizations at the organ and tissue levels. These studies provide a basis to clarify the role of particular anthocyanin species during abiotic stress.


Subject(s)
Anthocyanins/metabolism , Arabidopsis/physiology , Stress, Physiological , Absorption, Radiation , Arabidopsis/drug effects , Light , Magnesium Sulfate/pharmacology , Stress, Physiological/drug effects
7.
Planta ; 240(5): 931-40, 2014 Nov.
Article in English | MEDLINE | ID: mdl-24903357

ABSTRACT

MAIN CONCLUSION: Different abiotic stress conditions induce distinct sets of anthocyanins, indicating that anthocyanins have different biological functions, or that decoration patterns of each anthocyanin are used for unique purposes during stress. The induction of anthocyanin accumulation in vegetative tissues is often considered to be a response of plants to biotic or abiotic stress conditions. Arabidopsis thaliana (Arabidopsis) accumulates over 20 anthocyanins derived from the anthocyanidin cyanidin in an organ-specific manner during development, but the anthocyanin chemical diversity for their alleged stress protective functions remains unclear. We show here that, when grown in various abiotic stress conditions, Arabidopsis not only often accumulates significantly higher levels of total anthocyanins, but different stress conditions also favor the accumulation of different sets of anthocyanins. For example, the anthocyanin patterns of seedlings grown at pH 3.3 or in media lacking phosphate are very similar and characterized by relatively high levels of the anthocyanins A8 and A11. In contrast, anthocyanin inductive conditions (AIC) provided by high sucrose media are characterized by high accumulation of A9* and A5 relative to other stress conditions. The modifications present in each condition correlate reasonably well with the induction of the respective anthocyanin modification enzymes. Taken together, our results suggest that Arabidopsis anthocyanin profiles provide 'fingerprints' that reflect the stress status of the plants.


Subject(s)
Anthocyanins/biosynthesis , Arabidopsis Proteins/metabolism , Arabidopsis/metabolism , Stress, Physiological , Anthocyanins/chemistry , Anthocyanins/isolation & purification , Arabidopsis/drug effects , Arabidopsis/genetics , Arabidopsis Proteins/genetics , Cluster Analysis , Gene Expression Profiling , Gene Expression Regulation, Plant/drug effects , Hydrogen-Ion Concentration , Mannitol/pharmacology , Molecular Structure , Seedlings/genetics , Seedlings/metabolism , Sodium Chloride/pharmacology , Spectrophotometry
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