Pedreño García, María Ángeles

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Pedreño García, María Ángeles

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Pedreño García, María Ángeles

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Universidad de Murcia. Departamento de Biología Vegetal

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Open Access
Comparative study of metabolomic profile and antioxidant content of adult and In vitro leaves of Aristotelia chilensis
(MDPI, 2021-12-23) Crisóstomo Ayala, Karina Andrea; Pérez Manríquez, Claudia; Ferreres, Federico; Gil-izquierdo, Ángel; Hernández de la Torre, Martha; Sánchez Olate, Manuel; Ríos Leal, Darcy Graciela; Pedreño García, María Ángeles; Sabater Jara, Ana Belén; Biología Vegetal
This work aimed to identify the bioactive compounds present in adult maqui (Aristotelia chilensis) leaves from different stages of development and seasons of the year and compare them with leaves obtained from maqui plants grown in vitro. The qualitative and quantitative analysis of maqui leaf extracts by HPLC-DAD-ESI-MSn showed the presence of different polyphenolic compounds classified into galloyl and caffeoyl quinic acids, ellagitannins and ellagic acid- and flavonoid-derivatives. In general, the total phenolic content of the in vitro samples was higher than that of ex vitro samples, whereas the total flavonoid content was higher in winter basal leaves. Additionally, the analysis by HPLC-MS showed that the extract from spring basal leaves was enriched in quercetin, catechin, kaempferol and 3-caffeoyl quinic acids, while in the in vitro leaves extract, quercetin was not present. As regards lipophilic compounds identified by GC/MS, the samples of in vitro leaves showed a high presence of α-tocopherol and β-sitosterol. In contrast, the samples of adult leaves presented a hight level of linolenic and linoleic acids. These results suggest that maqui leaves could be an excellent source of antioxidants and lipophilic compounds for many industries, such as the nutraceutical and pharmaceutical industries.
Open Access
A smart strategy to Improve t-Resveratrol production in grapevine cells treated with Cyclodextrin polymers coated with magnetic nanoparticles
(MDPI, 2020-04-24) Gea Abellán, Alicia de; Rodríguez López, Marïa Isabel; Núñez Delicado, Estrella; Gabaldón, José Antonio; Almagro Romero, Lorena; Pedreño García, María Ángeles; Biología Vegetal
One of the most successfully procedures used to increase the production of t-resveratrol in Vitis vinifera suspension-cultured cells is the application of cyclodextrins (CDs) and methyl jasmonate (MJ) as elicitors. In particular, β-CDs are characterized by their chemical structure which makes them special, not only by acting as elicitors, but also because they are compounds capable of trapping high added-value hydrophobic molecules such as t-resveratrol. However, the use of β-CDs as elicitors increases the production costs of this compound, making their industrial exploitation economically unfeasible. Therefore, the development of β-CDs recovery strategies is necessary to provide a viable solution to their industrial use. In this work, carboxymethylated and hydroxypropylated β-CDs have been used to form polymers using epichlorohydrin (EPI) as a cross-linking agent. The polymers were coated to Fe3O4 nanoparticles and were jointly used with MJ to elicit V. vinifera suspension-cultured cells. Once elicitation experiments were finished, a magnet easily allowed the recovery of polymers, and t-resveratrol was extracted from them by using ethyl acetate. The results indicated that the production of t-resveratrol in the presence of free carboxymethyl-β-CDs was much lower than that found in the presence of carboxymethyl-β-cyclodextrins-EPI polymer coated magnetic nanoparticles. In addition, the maximal levels of t-resveratrol were found at 168 h of elicitation in the presence of 15 g/L hydroxypropyl-β-CDs polymer coated magnetic nanoparticles and MJ, and non-t-resveratrol was found in the extracellular medium, indicating that all the t-resveratrol produced by the cells and secreted into the culture medium was trapped by the polymer and extracted from it. This work also showed that polymers can be regenerated and reused during three cycles of continuous elicitation since the induction and adsorption capacity of hydroxypropyl-β-CDs polymer-coated magnetic nanoparticles after these cycles of elicitation remained high, allowing high concentrations of t-resveratrol to be obtained.
Open Access
Cyclodextrins increase phytosterol and tocopherol levels in suspension cultured cells obtained from mung beans and safflower
(Wiley, 2017-07-13) Tudela, Libertad Raquel; Almagro Romero, Lorena; Miras Moreno, Begoña; Pedreño García, María Ángeles; Sabater Jara, Ana Belén; Biología Vegetal
In this work, suspension-cultured cells of mung beans and safflower were used in order to analyze the effect of methyl jasmonate and/or cyclodextrins, on bioactive compound production such as phytosterols and tocopherols. The results indicated that mung bean suspension-cultured cells produced higher amount of total phytosterols and tocopherols. In particular, mung bean suspension-cultured cells produced almost 220-fold higher levels of tocopherols than safflower suspension-cultured cells in the best conditions. However, while cyclodextrins were able to enhance extracellular production of phytosterols, in the case of tocopherols, they only increased their intracellular accumulation. Our results showed that mung bean cells could be used as a highly efficient system for the production of phytosterols and tocopherols which have a wide range of biological activities.
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Induction of extracellular defense-related proteins in suspension cultured-cells of Daucus carota elicited with cyclodextrins and methyljasmonate
(Elsevier, 2014-02-18) Almagro Romero, Lorena; Pedreño García, María Ángeles; Sabater Jara, Ana Belén; Biología Vegetal
Suspension cultured-cells (SCC) of Daucus carota were used to evaluate the effect of methyl jasmonate and cyclodextrins, separately or in combination, on the induction of defense responses, particularly the accumulation of pathogenesis-related proteins. A comparative study of the extracellular proteome (secretome) between control and elicited carrot SCC pointed to the presence of amino acid sequences homologous to glycoproteins which have inhibitory activity against the cell-wall-degrading enzymes secreted by pathogens and/or are induced when carrot cells are exposed to a pathogen elicitor. Other amino acid sequences were homologous to Leucine-Rich Repeat domain-containing proteins, which play an essential role in defense against pathogens, as well as in the recognition of microorganisms, making them important players in the innate immunity of this plant. Also, some tryptic peptides were shown to be homologous to a thaumatin-like protein, showing high specificity to abiotic stress and to different reticuline oxidase-like proteins that displayed high levels of antifungal activity, suggesting that methyl jasmonate and cyclodextrins could play a role in mediating defense-related gene product expression in SCC of D. carota. Apart from these elicitor-inducible proteins, we observed the presence of PR-proteins in both control and elicited carrot SCC, suggesting that their expression is mainly constitutive. These PR-proteins are putative class IV chitinases, which also have inhibitory activity against pathogen growth and the class III peroxidases that participate in response to environmental stress (e.g. pathogen attack and oxidative), meaning that they are involved in defense responses triggered by both biotic and abiotic factors.
Open Access
Biotechnological approach to increase oxyresveratrol production in mulberry in vitro plants under elicitation
(MDPI, 2023-01-25) Nicolás Sánchez, Isabel; Almagro Romero, Lorena; Pedreño García, María Ángeles; Sabater Jara, Ana Belén; Biología Vegetal
Morus alba L. is used for a range of therapeutic purposes in Asian traditional medicine, and its extracts are reported to be effective against lipidemia, diabetes, and obesity, as well as being hepatoprotective and tyrosinase-inhibitory. They are also included in cosmetic products as anti-aging and skin-whitening agents. Stilbenes, the major bioactive compounds found in M. alba, have received renewed attention recently because of their putative activity against COVID-19. In this study M. alba plants were established in vitro, and the effect of elicitation on plant growth and stilbene accumulation, specifically oxyresveratrol and trans-resveratrol, was investigated. Different concentrations of the elicitors including methyl jasmonate and cyclodextrins were applied, and stilbene levels were determined in leaves, roots, and the culture medium. Elicitation of the M. alba plants with 5 mM cyclodextrins, alone or in combination with 10 µM methyl jasmonate, significantly increased the total phenolic content in the culture medium and leaves after 7 days of treatment. The higher total phenolic content in the roots of control plants and those treated only with methyl jasmonate indicated that cyclodextrins promoted metabolite release to the culture medium. Notably, the cyclodextrin-treated plants with the highest levels of oxy- and trans-resveratrol also had the highest total phenolic content and antioxidant capacity. These results indicate that elicited M. alba in vitro plants constitute a promising alternative source of bioactive stilbenes to supply pharmaceutical and cosmeceutical industries.
Metadata only
Caracterización de isoperoxidasas de Hipocotilos etiolados de Altramuz (L. Albus) / Mª Angeles Pedreño García ; directores Romualdo Muñoz Girón, Francisco García Carmona.
(Murcia : Universidad de Murcia, Departamento de Bioquímica y Biología Molecular,, 1987) Pedreño García, María Ángeles
Embargo
Production and localization of hydrogen peroxide and nitric oxide in grapevine cells elicited with cyclodextrins and methyl jasmonate
(Elsevier, 2019-06) Abellán Rubio, Marina; Almagro Romero, Lorena; Belchí Navarro, Sarai; Pedreño García, María Ángeles; Biología Vegetal
The use of methyl jasmonate, alone or in combination with cyclic oligosaccharides such as cyclodextrins, has proved to be a successful strategy for increasing the production of trans-resveratrol in Vitis vinifera cell cultures. However, understanding the intracellular signalling pathways involved in its production would improve the management of grapevine cells as biofactories of this high-value natural product. The results obtained herein conﬁrm the involvement of hydrogen peroxide and nitric oxide in cyclodextrins and methyl jasmonate-induced trans-resveratrol production in grapevine cell cultures. In fact, methyl jasmonate led to maximal intracellular levels of hydrogen peroxide and nitric oxide after 24 h of treatment, but extracellular hydrogen peroxide was only detected in the culture medium when grapevine cells were treated with cyclodextrins. The results derived from the cytochemical detection of H2O2 in elicited grapevine cell cultures also suggested that the combined treatment with cyclodextrins and methyl jasmonate not only increased the production of H2O2 but also released cell wall fragments with electron-dense deposits. Moreover, nitric oxide was localized in all the cellular compartments, particularly in the nucleus and cyto-plasmic organelles, whereas hydrogen peroxide was mainly found in cytoplasmic areas close to the cell wall, and in the nucleoplasm.
Embargo
Increased glucosinolate production in Brassica oleracea var. italica cell cultures due to coronatine activated genes involved in glucosinolate biosynthesis
(American Chemical Society, 2018-12-19) Sánchez Pujante, P.J.; Almagro Romero, Lorena; Belchí Navarro, Sarai; Pedreño García, María Ángeles; Sabater Jara, Ana Belén; Biología Vegetal
In this work, the effect of different elicitors and culture conditions on the production of glucosinolates in broccoli cell cultures was studied. The results showed that 0.5 μM coronatine was the best elicitor for increasing glucosinolate production (205-fold increase over untreated cells after 72 h of treatment). Furthermore, the expression levels of some genes related to the biosynthetic pathway of glucosinolates as well as three Myb transcription factors also have been studied. The highest glucosinolate levels found in coronatine-treated cells were closely correlated with the highest gene expression levels of Cyp79b2, Cyp83b1, St5a, Myb51, and Myb122 after 6 h of treatment. The data shown in this study provide new insight into the key metabolic steps involved in the biosynthesis of glucosinolates, which will be of use for future applications of metabolic engineering techniques in broccoli.
Restricted
Enhanced accumulation of phytosterols and phenolic compounds incyclodextrin-elicited cell suspension culture of Daucus carota
(Elsevier, 2016-06-15) Almagro Romero, Lorena; Miras Moreno, Begoña; Pedreño García, María Ángeles; Sabater Jara, Ana Belén; Biología Vegetal
Inthis work, suspension-culturedcells of Daucus carota wereusedto evaluate the effect of -cyclodextrins on the production of isoprenoid and phenolic compounds. The results showed that the phytosterols and phenolic compounds were accumulated in the extracellular medium (15100 g L−1 and 477.46 g L−1, respectively) in the presence of cyclodextrins. Unlike the phytosterol and phenolic compound content, -carotene (1138.03 g L−1), lutein (25949.54 g L−1) and -tocopherol (8063.82 g L−1) chlorophyll a (1625.13 g L−1) and b (9.958 (9958.33 g L−1) were mainly accumulated inside the cells. Therefore, cyclodextrins were able to induce the cytosolic mevalonate pathway, increasing the biosynthesis of phytosterols and phenolic compounds, and accumulate them outside the cells. However, in the absence of these cyclic oligosaccharidic elicitors, carrot cells mainly accumulated carotenoids through the methylerythritol 4-phosphate pathway. Therefore, the use of cyclodextrins would allow the extracellular accumulation of both phytosterols andphenolic compounds bydiverting the carbonflux towards the cytosolicmevalonate/phenylpropanoid pathway.
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Enhanced bioactive compound production in broccoli cells due to coronatine and methyl jasmonate is linked to antioxidative metabolism
(Elsevier, 2020-02-22) Sánchez-Pujante, Pedro Joaquín; Gionfriddo, Matteo; Díaz-Vivancos, Pedro; Almagro Romero, Lorena; Pedreño García, María Ángeles; Sabater Jara, Ana Belén; Biología Vegetal
Elicited broccoli suspension-cultured cells (SCC) provide a useful system for obtaining bioactive compounds, including glucosinolates (GS) and phenolic compounds (PCs). In this work, coronatine (Cor) and methyl jas monate (MJ) were used to increase the bioactive compound production in broccoli SCC. Although the use of Cor and MJ in secondary metabolite production has already been described, information concerning how elicitors affect cell metabolism is scarce. It has been suggested that Cor and MJ trigger defence reactions affecting the antioxidative metabolism. In the current study, the concentration of 0.5 μM Cor was the most effective treatment for increasing both the total antioxidant capacity (measured as ferulic acid equivalents) and glucosinolate content in broccoli SCC. The elicited broccoli SCC also showed higher polyphenol oxidase activity than the control cells. Elicitation altered the antioxidative metabolism of broccoli SCC, which displayed biochemical changes in antioxidant enzymes, a decrease in the glutathione redox state and an increase in lipid peroxidation levels. Furthermore, we studied the effect of elicitation on the protein profile and observed an induction of defence-related proteins. All of these findings suggest that elicitation not only increases bioactive compound production, but it also leads to mild oxidative stress in broccoli SCC that could be an important factor triggering the production of these compounds.