Pérez Gilabert, Manuela

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Pérez Gilabert, Manuela

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Pérez Gilabert, Manuela

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Universidad de Murcia. Departamento de Bioquímica y Biología MolecularA

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Purification and characterization of Terfezia claveryi TcCAT-1, a desert truffle catalase upregulated in mycorrhizal symbiosis
(Public Library of Science, 2019-07-10) Marqués Gálvez, José Eduardo; Morte Gómez, María Asunción; Navarro Ródenas, Alfonso; Pérez Gilabert, Manuela; García Carmona, Francisco; Biología Vegetal; Facultades de la UMU::Facultad de Biología
Terfezia claveryi Chatin is a mycorrhizal fungus that forms ectendomycorrhizal associations with plants of Helianthemum genus. Its appreciated edibility and drought resistance make this fungus a potential alternative crop in arid and semiarid areas of the Mediterranean region. In order to increase the knowledge about the biology of this fungus in terms of mycorrhiza formation and response to drought stress, a catalase from T. claveryi (TcCAT-1) has been purified to apparent homogeneity and biochemically characterized; in addition, the expression pattern of this enzyme during different stages of T. claveryi biological cycle and under drought stress conditions are reported. The results obtained, together with the phylogenetic analysis and homology modeling, indicate that TcCAT-1 is a homotetramer large subunit size monofunctional-heme catalase belonging to Clade 2. The highest expression of this enzyme occurs in mature mycorrhiza, revealing a possible role in mycorrhiza colonization, but it is not upregulated under drought stress. However, the H2O2 content of mycorrhizal plants submitted to drought stress is lower than in well watered treatments, suggesting that mycorrhization improves the plant’s oxidative stress response, although not via TcCAT-1 upregulation.
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Variación de la producción de etileno durante el crecimiento de hipocutilos etiolados de "lupinus albus L." : estudio microscópico del crecimiento /Manuela ´Perez Gilabert.
(Murcia : Universidad,, 1990) Pérez Gilabert, Manuela
Open Access
Desert truffle genomes reveal their reproductive modes and new insights into plant–fungal interaction and ectendomycorrhizal lifestyle
(Wiley, 2020-10-28) Marqués Gálvez, José Eduardo; Miyauchi, Shingo; Paolocci, Francesco; Navarro Ródenas, Alfonso; Arenas Jiménez, Francisco; Pérez Gilabert, Manuela; Morin, Emmanuelle; Auer, Lucas; Barry, Kerrie W.; Kuo, alan; Grigoriev, Igor V.; Martin, Francis M.; Kohler, Annegret; Morte Gómez, María Asunción; Biología Vegetal; Facultades de la UMU::Facultad de Biología
Desert truffles are edible hypogeous fungi forming ectendomycorrhizal symbiosis with plants of Cistaceae family. Knowledge about the reproductive modes of these fungi and the molecular mechanisms driving the ectendomycorrhizal interaction is lacking. Genomes of the highly appreciated edible desert truffles Terfezia claveryi Chatin and Tirmania nivea Trappe have been sequenced and compared with other Pezizomycetes. Transcriptomes of T. claveryi × Helianthemum almeriense mycorrhiza from well-watered and drought-stressed plants, when intracellular colonizations is promoted, were investigated. We have identified the fungal genes related to sexual reproduction in desert truffles and desert-truffles-specific genomic and secretomic features with respect to other Pezizomycetes, such as the expansion of a large set of gene families with unknown Pfam domains and a number of species or desert-truffle-specific small secreted proteins differentially regulated in symbiosis. A core set of plant genes, including carbohydrate, lipid-metabolism, and defence-related genes, differentially expressed in mycorrhiza under both conditions was found. Our results highlight the singularities of desert truffles with respect to other mycorrhizal fungi while providing a first glimpse on plant and fungal determinants involved in ecto to endo symbiotic switch that occurs in desert truffle under dry conditions.
Open Access
Mycelium of Terfezia claveryi as inoculum source to produce desert truffle mycorrhizal plants
(Springer, 2018-09-20) Navarro-Ródenas, Alfonso; Chávez, Daniel; Gutiérrez, Almudena; Arenas Jiménez, Francisco; Morte Gómez, María Asunción; Pérez Gilabert, Manuela; Biología Vegetal
Terfezia claveryi Chatin was the first desert truffle species to be cultivated, the mycorrhizal plants being successfully produced by using both desert truffle spores and mycelia. However, it is more advisable to use mycelium than spores whenever possible and profitable. Given the low yields of mycelia obtained using traditional culture methods of this truffle, the medium composition was modified in an attempt to determine its nutritional requirements. For this, an assay involving response surface methodology was performed using Box-Behnken design to find the optimal parameters for the high production of mycelial biomass. The best results were obtained with glucose as carbon source, buffering the pH at 5 during culture, adding a pool of vitamins and adjusting the optimal concentrations of carbon and nitrogen sources of the MMN medium. Biomass production increased from 0.3 to 3 g L-1 dry weight and productivity increased from 10.7 to 95.8 mg L-1 d-1 dry weight. The produced mycelium was able to colonize Helianthemum roots efficiently, providing more than 50 % ectomycorrhizal colonization.
Open Access
The first comprehensive phylogenetic and biochemical analysis of NADH diphosphatases reveals that the enzyme from Tuber melanosporum is highly active towards NAD+
(Nature Research, 2019) García-Saura, Antonio Ginés; Zapata-Pérez, Rubén; Martínez-Moñino, Ana Belén; Francisco Hidalgo, José; Morte Gómez, María Asunción; Pérez Gilabert, Manuela; Sánchez Ferrer, Álvaro; Bioquímica y Biología Molecular A
Las hidrolasas Nudix (por nucleósido difosfatasas unidas a otros motivos, X) son una familia diversa de proteínas capaces de escindir una enorme variedad de sustratos, que van desde azúcares nucleotídicos hasta ARNs coronados con NAD+. Aunque todos los miembros de esta superfamilia comparten un motivo catalítico común conservado, la caja Nudix, su especificidad de sustrato radica en rasgos de secuencia específicos, que dan lugar a diferentes subfamilias. Entre ellas, las NADH pirofosfatasas o difosfatasas (NADDs) están poco estudiadas y no se sabe nada de su distribución. Para solucionar esto, diseñamos un patrón compatible con Prosite para identificar nuevas secuencias de NADDs. El escaneo in silico de la base de datos UniProtKB mostró que el 3% de las proteínas Nudix eran NADDs y mostraban 21 arquitecturas de dominio diferentes, siendo la arquitectura canónica (NUDIX-like_zf-NADH-PPase_NUDIX) la más abundante (53%). Curiosamente, las secuencias fúngicas de NADD destacaban entre los eucariotas y se distribuían en varias clases, incluyendo los Pezizomicetos. Inesperadamente, en esta última Clase de hongos, se encontró que los NADDs estaban presentes desde el ancestro reciente más común hasta las Tuberaceae, siguiendo una distribución de la filogenia molecular similar a la descrita previamente usando dos mil genes individuales concatenados. Por último, cuando se caracterizó bioquímicamente la NADD de Tuber melanosporum, formadora de trufas, ésta mostró la mayor relación de eficiencia catalítica NAD+/NADH jamás descrita. .
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Actividades catalíticas de lipoxigenasa en medio acuoso y micelar /Manuela Perez Gilabert ; directores Francisco García Carmona, Alvaro Sánchez Ferrer
(Murcia : Universidad de Murcia, Facultad de Biologia,, 1994) Pérez Gilabert, Manuela
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A continuous spectrophotometric assay for determination of the aureusidin synthase activity of tyrosinase
(Wiley, 2010-04-13) Jiménez-Atiénzar, Mercedes; Cabanes Cos, Juana; Escribano Cebrián, Josefa; Gandía Herrero, Fernando; Pérez Gilabert, Manuela; García Carmona, Francisco; Departamento de Bioquímica y Biología Molecular "A"
Introduction – Aurones (aureusidin glycosides) are plant flavonoids that provide yellow colour to the flowers of some orna-mental plants. In this study we analyse the capacity of tyrosinase to catalyse the synthesis of aureusidin by tyrosinase fromthe chalcone THC (2′,4′,6′,4–tetrahydroxychalcone).Objective – To develop a simple continuous spectrophotometric assay for the analysis of the spectrophotometric and kineticcharacteristics of THC oxidation by tyrosinase.Methodology – THC oxidation was routinely assayed by measuring the increase in absorbance at 415 nm vs. reaction time.Results – According to the mechanism proposed for tyrosinase, the enzymatic reaction involves the o-hydroxylation of themonophenol THC to the o-diphenol (PHC, 2′,4′,6′,3,4 – pentahydroxychalcone), which is then oxidised to the correspondingo-quinone in a second enzymatic step. This product is highly unstable and thus undergoes a series of fast chemical reactionsto produce aureusidin. In these experimental conditions, the optimum pH for THC oxidation is 4.5. The progress curvesobtained for THC oxidation showed the appearance of a lag period. The following kinetic parameters were also determined:Km = 0.12 mM , Vm = 13 mM /min, Vm/Km = 0.11/min.Conclusion – This method has made it possible to analyse the spectrophotometric and kinetic characteristics of THC by tyro-sinase. This procedure has the advantages of a short analysis time, straightforward measurement techniques and repro-ducibility. In addition, it also allows the study of tyrosinase inhibitors, such as tropolone.
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Basic and Applied Research for Desert Truffle Cultivation
(Springer, 2017) Morte Gómez, María Asunción; Pérez Gilabert, Manuela; Gutiérrez Abbad, Almudena; Arenas Jiménez, Francisco; Marqués Gálvez, José Eduardo; Bordallo, Juan Julián; Rodríguez, Antonio; Berná Amorós, Luis Miguel; Lozano-Carrillo, Cecilia; Navarro Ródenas, Alfonso; Biología Vegetal
This chapter summarizes the latest basic and applied advances in desert truffle research carried out to improve our knowledge of the biodiversity, physiology, biotechnology, and cultivation of these hypogeous and edible fungi. ITS-rDNA sequences in phylo-geographic studies and host plant and soil pH characteristics have been the key to describing eight new desert truffle species. The production of desert truffle mycorrhizal plants has been improved by using β-cyclodextrin and bioreactors for mycelium culture and native beneficial bacteria (PGPR and MHB) to increase seedling survival and mycorrhization. Some fungal enzymes have also been characterized in Terfezia claveryi ascocarps. The presence of alkaline phosphatase both in mycelia and ascocarps indicates that this enzyme plays an important role during the life cycle of T. claveryi, while acid phosphatase might be involved in a process that takes place during the ascocarp stage. Numerous desert truffle plantations have been established in Spain in the last 10 years. A high density of mycorrhizal plants combined with a proper irrigation are two important factors to stimulate ascocarp production. The combination of a high rate of intracellular colonization together with the fine-tuned expression of fungal and plant aquaporins could result in a morpho-physiological adaptation of this symbiosis in drought conditions. Moreover, desert truffle sylviculture is proposed for improving truffle production and for conserving the natural areas where desert truffle grow.