scholarly journals UGT84F9 is the major flavonoid UDP-glucuronosyltransferase in Medicago truncatula

2021 ◽  
Author(s):  
Olubu A Adiji ◽  
Maite L Docampo-Palacios ◽  
Anislay Alvarez-Hernandez ◽  
Giulio M Pasinetti ◽  
Xiaoqiang Wang ◽  
...  
Keyword(s):  
Medicago Truncatula ◽  
Phase Ii ◽  
Loss Of Function ◽  
In Planta ◽  
Model Legume ◽  
Flavonoid Compounds ◽  
Wide Range ◽  
Udp Glucuronosyltransferase ◽  
Phase Ii Metabolites

Abstract Mammalian phase II metabolism of dietary plant flavonoid compounds generally involves substitution with glucuronic acid. In contrast, flavonoids mainly exist as glucose conjugates in plants, and few plant UDP-glucuronosyltransferase enzymes have been identified to date. In the model legume Medicago truncatula, the major flavonoid compounds in the aerial parts of the plant are glucuronides of the flavones apigenin and luteolin. Here we show that the M. truncatula glycosyltransferase UGT84F9 is a bi-functional glucosyl/glucuronosyl transferase in vitro, with activity against a wide range of flavonoid acceptor molecules including flavones. However, analysis of metabolite profiles in leaves and roots of M. truncatula ugt84f9 loss of function mutants revealed that the enzyme is essential for formation of flavonoid glucuronides, but not most flavonoid glucosides, in planta. We discuss the use of plant UGATs for the semi-synthesis of flavonoid phase II metabolites for clinical studies.

scholarly journals Identification of a putative DNA-binding protein in Arabidopsis that acts as a susceptibility hub and interacts with multiple Pseudomonas syringae effectors

2020 ◽  
Author(s):  
Karl Schreiber ◽  
Jennifer D Lewis
Keyword(s):  
Pseudomonas Syringae ◽  
Transcriptional Profiling ◽  
Effector Proteins ◽  
Ribosomal Protein Genes ◽  
Loss Of Function ◽  
Wild Type ◽  
In Planta ◽  
Arabidopsis Protein ◽  
Wide Range

Phytopathogens use secreted effector proteins to suppress host immunity and promote pathogen virulence, and there is increasing evidence that the host-pathogen interactome comprises a complex network. In an effort to identify novel interactors of the Pseudomonas syringae effector HopZ1a, we performed a yeast two-hybrid screen that identified a previously uncharacterized Arabidopsis protein that we designate HopZ1a Interactor 1 (ZIN1). Additional analyses in yeast and in planta revealed that ZIN1 also interacts with several other P. syringae effectors. We show that an Arabidopsis loss-of-function zin1 mutant is less susceptible to infection by certain strains of P. syringae, while overexpression of ZIN1 results in enhanced susceptibility. Functionally, ZIN1 exhibits topoisomerase-like activity in vitro. Transcriptional profiling of wild-type and zin1 Arabidopsis plants inoculated with P. syringae indicated that while ZIN1 regulates a wide range of pathogen-responsive biological processes, the list of genes more highly expressed in zin1 versus wild-type plants was particularly enriched for ribosomal protein genes. Altogether, these data illuminate ZIN1 as a potential susceptibility hub that interacts with multiple effectors to influence the outcome of plant-microbe interactions.

scholarly journals Efficacy of Variable Tetraconazole Rates Against Cercospora beticola Isolates with Differing In Vitro Sensitivities to DMI Fungicides

Plant Disease ◽  
2012 ◽  
Vol 96 (12) ◽  
pp. 1749-1756 ◽  
Author(s):  
Melvin D. Bolton ◽  
Viviana Rivera-Varas ◽  
Luis E. del Río Mendoza ◽  
Mohamed F. R. Khan ◽  
Gary A. Secor
Keyword(s):  
Sugar Beet ◽  
Disease Severity ◽  
Cercospora Beticola ◽  
In Planta ◽  
Sensitivity Testing ◽  
In Vitro Sensitivity ◽  
Wide Range ◽  
Fungicide Efficacy ◽  
Dmi Fungicides

Cercospora leaf spot (CLS) of sugar beet is caused by the fungus Cercospora beticola. CLS management practices include the application of the sterol demethylation inhibitor (DMI) fungicides tetraconazole, difenoconazole, and prothioconazole. Evaluating resistance to DMIs is a major focus for CLS fungicide resistance management. Isolates were collected in 1997 and 1998 (baseline sensitivity to tetraconazole, prothioconazole, or difenoconazole) and 2007 through 2010 from the major sugar-beet-growing regions of Minnesota and North Dakota and assessed for in vitro sensitivity to two or three DMI fungicides. Most (47%) isolates collected in 1997–98 exhibited 50% effective concentration (EC50) values for tetraconazole of <0.01 μg ml–1, whereas no isolates could be found in this EC50 range in 2010. Since 2007, annual median and mean tetraconazole EC50 values have generally been increasing, and the frequency of isolates with EC50 values >0.11 μg ml–1 increased from 2008 to 2010. In contrast, the frequency of isolates with EC50 values for prothioconazole of >1.0 μg ml–1 has been decreasing since 2007. Annual median difenoconazole EC50 values appears to be stable, although annual mean EC50 values generally have been increasing for this fungicide. Although EC50 values are important for gauging fungicide sensitivity trends, a rigorous comparison of the relationship between in vitro EC50 values and loss of fungicide efficacy in planta has not been conducted for C. beticola. To explore this, 12 isolates exhibiting a wide range of tetraconazole EC50 values were inoculated to sugar beet but no tetraconazole was applied. No relationship was found between isolate EC50 value and disease severity. To assess whether EC50 values are related to fungicide efficacy in planta, sugar beet plants were sprayed with various dilutions of Eminent, the commercial formulation of tetraconazole, and subsequently inoculated with isolates that exhibited very low, medium, or high tetraconazole EC50 values. The high EC50 isolate caused significantly more disease than isolates with medium or very low EC50 values at the field application rate and most reduced rates. Because in vitro sensitivity testing is typically carried out with the active ingredient of the commercial fungicide, we investigated whether loss of disease control was the same for tetraconazole as for the commercial product Eminent. The high EC50 isolate caused more disease on plants treated with tetraconazole than Eminent but disease severity was not different between plants inoculated with the very low EC50 isolate.

scholarly journals LeGOO: An Expertized Knowledge Database for the Model Legume Medicago truncatula

2019 ◽  
Vol 61 (1) ◽  
pp. 203-211 ◽  
Author(s):  
S�bastien Carr�re ◽  
Marion Verdenaud ◽  
Clare Gough ◽  
J�r�me Gouzy ◽  
Pascal Gamas
Keyword(s):  
Medicago Truncatula ◽  
Reverse Genetics ◽  
Arbuscular Mycorrhizal ◽  
Biological Information ◽  
Model Species ◽  
Model Legume ◽  
Wide Range ◽  
Legume Symbiosis ◽  
Symbiotic Nodule ◽  
Set Up

Abstract Medicago truncatula was proposed, about three decades ago, as a model legume to study the Rhizobium-legume symbiosis. It has now been adopted to study a wide range of biological questions, including various developmental processes (in particular root, symbiotic nodule and seed development), symbiotic (nitrogen-fixing and arbuscular mycorrhizal endosymbioses) and pathogenic interactions, as well as responses to abiotic stress. With a number of tools and resources set up in M. truncatula for omics, genetics and reverse genetics approaches, massive amounts of data have been produced, as well as four genome sequence releases. Many of these data were generated with heterogeneous tools, notably for transcriptomics studies, and are consequently difficult to integrate. This issue is addressed by the LeGOO (for Legume Graph-Oriented Organizer) knowledge base (https://www.legoo.org), which finds the correspondence between the multiple identifiers of the same gene. Furthermore, an important goal of LeGOO is to collect and represent biological information from peer-reviewed publications, whatever the technical approaches used to obtain this information. The information is modeled in a graph-oriented database, which enables flexible representation, with currently over 200,000 relations retrieved from 298 publications. LeGOO also provides the user with mining tools, including links to the Mt5.0 genome browser and associated information (on gene functional annotation, expression, methylome, natural diversity and available insertion mutants), as well as tools to navigate through different model species. LeGOO is, therefore, an innovative database that will be useful to the Medicago and legume community to better exploit the wealth of data produced on this model species.

scholarly journals Plant biotransformation of T2 and HT2 toxin in cultured organs of Triticum durum Desf

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Laura Righetti ◽  
Tania Körber ◽  
Enrico Rolli ◽  
Gianni Galaverna ◽  
Michele Suman ◽  
...  
Keyword(s):  
Triticum Durum ◽  
In Planta ◽  
Plant Organs ◽  
Uninfected Plant ◽  
Complete Absorption ◽  
Triticum Durum Desf ◽  
Phase Ii Metabolites ◽  
T2 Toxin ◽  
First Time

Abstract The present study aimed at elucidating the uptake and biotransformation of T2 and HT2 toxins in five cultivars of durum wheat, by means of cultured plant organs. An almost complete absorption of T2 toxin (up to 100 µg) was noticed after 7 days, along with the contemporaneous formation of HT2 in planta, whereas HT2 showed a slower uptake by uninfected plant organs. Untargeted MS-analysis allowed to identify a large spectrum of phase I and phase II metabolites, resulting in 26 T2 and 23 HT2 metabolites plus tentative isomers. A novel masked mycotoxin, 3-acetyl-HT2-glucoside, was reported for the first time in wheat. The in vitro approach confirmed its potential to both investigate the contribution of plant metabolism in the biosynthesis of masked mycotoxins and to foresee the development of biocatalytic tools to develop nature-like mixtures to be used as reference materials.

Cloning, molecular characterization, and expression analysis of several red clover cDNAs

2006 ◽  
Vol 86 (2) ◽  
pp. 465-468
Author(s):  
Michael L. Sullivan ◽  
Sharon L. Thoma
Keyword(s):  
Medicago Truncatula ◽  
Expression Analysis ◽  
Binding Protein ◽  
Sequence Similarity ◽  
Expression Patterns ◽  
Molecular Genetic ◽  
Red Clover ◽  
Model Legume ◽  
Gene Set ◽  
Wide Range

To begin gathering information regarding nucleotide sequence similarity between red clover genes and other plant species, especially the model legume Medicago truncatula, several random red clover cDNAs were sequenced. The analyzed cDNAs included genes encoding actin; several proteins involved in photosynthesis including PsaH, PsbR, PsbX, early light-induced protein (ELIP), ferredoxin, chlorophyll a/b binding protein; fructose-bisphosphate aldolase;chloroplastic superoxide dismutase; and GTP-binding protein typA. The gene set had a median sequence identity of 92% with their counterparts from M. truncatula, suggesting its available genomics tools can be applied to red clover. An expression analysis of the gene set in various red clover tissues indicates the genes show a wide range of expression patterns. Consequently, this set of cDNAs and associated data are proving useful as controls in molecular genetic experiments involving red clover. Key words: Red clover, Trifolium pratense, Medicago truncatula, forage legume, genomics, inquiry-based learning

scholarly journals The Redox Cofactor F420Protects Mycobacteria from Diverse Antimicrobial Compounds and Mediates a Reductive Detoxification System

2016 ◽  
Vol 82 (23) ◽  
pp. 6810-6818 ◽  
Author(s):  
Thanavit Jirapanjawat ◽  
Blair Ney ◽  
Matthew C. Taylor ◽  
Andrew C. Warden ◽  
Shahana Afroze ◽  
...  
Keyword(s):  
Antimicrobial Resistance ◽  
Mycobacterium Smegmatis ◽  
Antimicrobial Compounds ◽  
Loss Of Function ◽  
Detoxifying Enzymes ◽  
Content Type ◽  
Wide Range ◽  
Detoxification System ◽  
Mutant Strains

ABSTRACTA defining feature of mycobacterial redox metabolism is the use of an unusual deazaflavin cofactor, F420. This cofactor enhances the persistence of environmental and pathogenic mycobacteria, including after antimicrobial treatment, although the molecular basis for this remains to be understood. In this work, we explored our hypothesis that F420enhances persistence by serving as a cofactor in antimicrobial-detoxifying enzymes. To test this, we performed a series of phenotypic, biochemical, and analytical chemistry studies in relation to the model soil bacteriumMycobacterium smegmatis. Mutant strains unable to synthesize or reduce F420were found to be more susceptible to a wide range of antibiotic and xenobiotic compounds. Compounds from three classes of antimicrobial compounds traditionally resisted by mycobacteria inhibited the growth of F420mutant strains at subnanomolar concentrations, namely, furanocoumarins (e.g., methoxsalen), arylmethanes (e.g., malachite green), and quinone analogues (e.g., menadione). We demonstrated that promiscuous F420H2-dependent reductases directly reduce these compounds by a mechanism consistent with hydride transfer. Moreover,M. smegmatisstrains unable to make F420H2lost the capacity to reduce and detoxify representatives of the furanocoumarin and arylmethane compound classes in whole-cell assays. In contrast, mutant strains were only slightly more susceptible to clinical antimycobacterials, and this appeared to be due to indirect effects of F420loss of function (e.g., redox imbalance) rather than loss of a detoxification system. Together, these data show that F420enhances antimicrobial resistance in mycobacteria and suggest that one function of the F420H2-dependent reductases is to broaden the range of natural products that mycobacteria and possibly other environmental actinobacteria can reductively detoxify.IMPORTANCEThis study reveals that a unique microbial cofactor, F420, is critical for antimicrobial resistance in the environmental actinobacteriumMycobacterium smegmatis. We show that a superfamily of redox enzymes, the F420H2-dependent reductases, can reduce diverse antimicrobialsin vitroandin vivo.M. smegmatisstrains unable to make or reduce F420become sensitive to inhibition by these antimicrobial compounds. This suggests that mycobacteria have harnessed the unique properties of F420to reduce structurally diverse antimicrobials as part of the antibiotic arms race. The F420H2-dependent reductases that facilitate this process represent a new class of antimicrobial-detoxifying enzymes with potential applications in bioremediation and biocatalysis.

scholarly journals A Polyamine Oxidase from Selaginella lepidophylla (SelPAO5) can Replace AtPAO5 in Arabidopsis through Converting Thermospermine to Norspermidine instead to Spermidine

Plants ◽  
2019 ◽  
Vol 8 (4) ◽  
pp. 99 ◽  
Author(s):  
G. H. M. Sagor ◽  
Tomonobu Kusano ◽  
Thomas Berberich
Keyword(s):  
Normal Values ◽  
Normal Growth ◽  
Polyamine Oxidase ◽  
Loss Of Function ◽  
Wild Type ◽  
In Planta ◽  
Selaginella Lepidophylla ◽  
Growth Phenotype ◽  
Polyamine Oxidases

Of the five polyamine oxidases in Arabidopsis thaliana, AtPAO5 has a substrate preference for the tetraamine thermospermine (T-Spm) which is converted to triamine spermidine (Spd) in a back-conversion reaction in vitro. A homologue of AtPAO5 from the lycophyte Selaginella lepidophylla (SelPAO5) back-converts T-Spm to the uncommon polyamine norspermidine (NorSpd) instead of Spd. An Atpao5 loss-of-function mutant shows a strong reduced growth phenotype when growing on a T-Spm containing medium. When SelPAO5 was expressed in the Atpao5 mutant, T-Spm level decreased to almost normal values of wild type plants, and NorSpd was produced. Furthermore the reduced growth phenotype was cured by the expression of SelPAO5. Thus, a NorSpd synthesis pathway by PAO reaction and T-Spm as substrate was demonstrated in planta and the assumption that a balanced T-Spm homeostasis is needed for normal growth was strengthened.

scholarly journals Comparative studies of urolithins and their phase II metabolites on macrophage and neutrophil functions

2020 ◽  
Author(s):  
Aneta Bobowska ◽  
Sebastian Granica ◽  
Agnieszka Filipek ◽  
Matthias F. Melzig ◽  
Thomas Moeslinger ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Inflammatory Response ◽  
Phase Ii ◽  
Comparative Studies ◽  
Active Metabolite ◽  
Phase Ii Metabolism ◽  
Urolithin A ◽  
Tnf Α ◽  
Phase Ii Metabolites

Abstract Purpose Ellagitannins are high molecular weight polyphenols present in high quantities in various food products. They are metabolized by human and animal gut microbiota to postbiotic metabolites-urolithins, bioavailable molecules of a low molecular weight. Following absorption in the gut, urolithins rapidly undergo phase II metabolism. Thus, to fully evaluate the mechanisms of their biological activity, the in vitro studies should be conducted for their phase II conjugates, mainly glucuronides. The aim of the study was to comparatively determine the influence of urolithin A, iso-urolithin A, and urolithin B together with their respective glucuronides on processes associated with the inflammatory response. Methods The urolithins obtained by chemical synthesis or isolation from microbiota cultures were tested with their respective glucuronides isolated from human urine towards modulation of inflammatory response in THP-1-derived macrophages, RAW 264.7 macrophages, PBMCs-derived macrophages, and primary neutrophils. Results Urolithin A was confirmed to be the most active metabolite in terms of LPS-induced inflammatory response inhibition (TNF-α attenuation, IL-10 induction). The observed strong induction of ERK1/2 phosphorylation has been postulated as the mechanism of its action. None of the tested glucuronide conjugates was active in terms of pro-inflammatory TNF-α inhibition and anti-inflammatory IL-10 and TGF-β1 induction. Conclusion Comparative studies of the most abundant urolithins and their phase II conjugates conducted on human and murine immune cells unambiguously confirmed urolithin A to be the most active metabolite in terms of inhibition of the inflammatory response. Phase II metabolism was shown to result in the loss of urolithins’ pharmacological properties.

scholarly journals Loss of the nodule-specific cysteine rich peptide, NCR169, abolishes symbiotic nitrogen fixation in the Medicago truncatula dnf7 mutant

2015 ◽  
Vol 112 (49) ◽  
pp. 15232-15237 ◽  
Author(s):  
Beatrix Horváth ◽  
Ágota Domonkos ◽  
Attila Kereszt ◽  
Attila Szűcs ◽  
Edit Ábrahám ◽  
...  
Keyword(s):  
Nitrogen Fixation ◽  
Medicago Truncatula ◽  
Symbiotic Nitrogen Fixation ◽  
Root Nodules ◽  
Nitrogen Fixing ◽  
In Planta ◽  
Functional Roles ◽  
Absolute Requirement ◽  
Cell Layers

Host compatible rhizobia induce the formation of legume root nodules, symbiotic organs within which intracellular bacteria are present in plant-derived membrane compartments termed symbiosomes. In Medicago truncatula nodules, the Sinorhizobium microsymbionts undergo an irreversible differentiation process leading to the development of elongated polyploid noncultivable nitrogen fixing bacteroids that convert atmospheric dinitrogen into ammonia. This terminal differentiation is directed by the host plant and involves hundreds of nodule specific cysteine-rich peptides (NCRs). Except for certain in vitro activities of cationic peptides, the functional roles of individual NCR peptides in planta are not known. In this study, we demonstrate that the inability of M. truncatula dnf7 mutants to fix nitrogen is due to inactivation of a single NCR peptide, NCR169. In the absence of NCR169, bacterial differentiation was impaired and was associated with early senescence of the symbiotic cells. Introduction of the NCR169 gene into the dnf7-2/NCR169 deletion mutant restored symbiotic nitrogen fixation. Replacement of any of the cysteine residues in the NCR169 peptide with serine rendered it incapable of complementation, demonstrating an absolute requirement for all cysteines in planta. NCR169 was induced in the cell layers in which bacteroid elongation was most pronounced, and high expression persisted throughout the nitrogen-fixing nodule zone. Our results provide evidence for an essential role of NCR169 in the differentiation and persistence of nitrogen fixing bacteroids in M. truncatula.

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