Belchí Navarro, Sarai

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Belchí Navarro, Sarai

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Belchí Navarro, Sarai

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Universidad de Murcia. Departamento de Biología Vegetal

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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.
Open Access
Caracterización de la producción de resveratrol y análisis de proteínas en cultivos celulares de Vitis vinífera cv Monastrell
(2012-11-02) Belchí Navarro, Sarai; Pedreño García, María Ángeles; Pedreño García, María Ángeles; Departamento de Biología Vegetal
En esta memoria de investigación se han utilizado cultivos celulares de Vitis vinifera cv Monastrell para la producción de trans-resveratrol. En este sentido se ha realizado la caracterización del crecimiento de suspensiones celulares de Monastrell en diferentes medios minerales y concentraciones de sacarosa. Asimismo se caracterizó la producción extracelular de trans-resveratrol mediante la utilización de distintos elicitores tales como ciclodextrinas de distinta naturaleza, jasmonato de metilo, luz UV, fosetil de aluminio, ácido salicílico y etefón. Asimismo, se realizó el estudio de la implicación del calcio, del peróxido de hidrógeno, el óxido nítrico así como los eventos de fosforilación y desfosforilación de proteínas en los fenómenos de señalización desencadenados por la elicitación de suspensiones celulares de Monastrell que conducen a la producción de trans-resveratrol. Finalmente, se realizó el análisis de las proteínas extracelulares tanto en condiciones control como elicitadas con distintos compuestos con el fin de determinar las respuestas de defensa inducidas en estas condiciones. In this memory Vitis vinifera cv Monastrell cell cultures were used to produce trans-resveratrol. The growth of Monastrell cell cultures were characterized using different mineral media and sucrose concentrations. Extracellular production of trans-resveratrol was also characterized using different elicitors such as different nature cyclodextrins, methyl jasmonate, UV light, fosetyl aluminum, salicylic acid and ethephon. Moreover it was study the involvement of calcium, hydrogen peroxide, nitric oxide and phosphorylation and dephosphorylation of proteins in the signaling events triggered by elicitation of Monastrell cell cultures leading to trans-resveratrol production. Finally, it was analyzed extracellular proteins in control and elicited cell cultures in the presence of different elicitors to determine defense responses induced in these conditions.
Open Access
Resveratrol lacks antifungal activity against Candida albicans.
(Springer, 2012-03-30 ) Collado-González, Mar; Guirao-Abad, José Pedro; Sánchez-Fresneda Pinto, Ruth; Belchí Navarro, Sarai; Argüelles Ordóñez, Juan Carlos; Genética y Microbiología
The putative candicidal activity of resveratrol is currently a matter of controversy. Here, the antifungal activity as well as the antioxidant response of resveratrol against Candida albicans, have been tested in a set of strains with a well-established genetic background At the doses usually employed in antifungal tests (10–40 lg/ml), resveratrol has no effect on the exponential growth of the C. albicans CAI.4 strain, a tenfold increase (400 lg/ml) was required in order to record a certain degree of cell killing, which was negligible in comparison with the strong antifungal effect caused by the addition of amphotericin B (5 lg/ml). An identical pattern was recorded in the prototrophic strains of C. albicans SC5314 and RM-100, whereas the oxidative sensitive trehalose-deficient mutant (tps1/tps1 strain) was totally refractory to the presence of resveratrol. In turn, the serum-induced yeast-to-hypha transition remained unaffected upon addition of different concentrations of resveratrol. Determination of endogenous trehalose and catalase activity, two antioxidant markers in C. albicans; revealed no significant changes in their basal contents induced by resveratrol. Collectively, our results seem to dismiss a main antifungal role as well as the therapeutic application of resveratrol against the infections caused by C. albicans.
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Induction of sesquiterpenes, phytoesterols and extracellular pathogenesis-related proteins in elicited cell cultures of Capsicum annuum
(Elsevier, 2010-07-01) Ferrer Ayala, María Angeles; Ros Barceló, Alfonso; Almagro Romero, Lorena; Belchí Navarro, Sarai; Pedreño García, María Ángeles; Sabater Jara, Ana Belén; Biología Vegetal
Capsicum annuum suspension cell cultures were used to evaluate the effect of cyclodextrins and methyl jasmonate as elicitors of defence responses. The induced defence responses included the accumulation of sesquiterpenes and phytosterols and the activation of pathogenesis-related proteins, leading to reinforcement and modification of the cell wall architecture during elicitation and protection cells against biotic stress. The results showed that the addition of both cyclodextrins and methyl jasmonate induced the biosynthesis of two sesquiterpenes, aromadendrene and solavetivone. This response was clearly synergistic since the increase in the levels of these compounds was much greater in the presence of both elicitors than when they were used separately. The biosynthesis of phytosterols was also induced in the combined treatment, as the result of an additive effect. Likewise, the exogenous application of methyl jasmonate induced the accumulation of pathogenesis-related proteins. The analysis of the extracellular proteome showed the presence of amino acid sequences homologous to PR1 and 4, NtPRp27-like proteins and class I chitinases, peroxidases and the hydrolytic enzymes LEXYL1 and 2, arabinosidases, pectinases, nectarin IV and leucin-rich repeat protein, which suggests that methyl jasmonate plays a role in mediating defence-related gene product expression in C. annuum. Apart from these methyl jamonate-induced proteins, other PR proteins were found in both the control and elicited cell cultures of C. annuum. These included class IV chitinases, -1,3-glucanases, thaumatin-like proteins and peroxidases, suggesting that their expression is mainly constitutive since they are involved in growth, development and defence processes.
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Recent trends in the biotechnological production of tocopherols using in vitro cultures
(Springer, 2021-03-27) Almagro Romero, Lorena; Belchí Navarro, Sarai; Pedreño García, María Ángeles; Sabater Jara, Ana Belén; Biología Vegetal
Tocopherols, a class of terpenoid compounds with vitamin-E activity, are produced by photosynthetic organisms, including cyanobacteria, algae, and plants. The chemical structure of tocopherols consists of a hydrophobic prenyl tail linked to a polar chromanol head. Depending on the position and number of methyl groups on the chromanol head, tocopherols are classed as a, b, c, and d isomers. Tocopherols play a key role in plant defense, and have bioactive properties beneficial for human health, being reported to prevent atherosclerosis, Parkinson’s and Alzheimer’s diseases, eyesight degeneration, diabetes, and coronary heart disease. The high market demand for tocopherols calls for new strategies to improve their supply from sustainable sources, and a promising approach is to develop biotechnological factories with tocopherol-producing plant and microalgae in vitro cultures. This review provides a detailed overview of different strategies devised to enhance tocopherol production using such systems, with particular focus on elicitation and feeding, culture conditions, and the application of metabolic engineering.
Open Access
Changes in the secretome of Vitis vinifera cv. Monastrell cell cultures treated with cyclodextrins and methyl jasmonate
(ELSEVIER FRANCE-EDITIONS SCIENTIFIQUES MEDICALES ELSEVIER, 2019-02) Bru, R.; Almagro Romero, Lorena; Belchí Navarro, Sarai; Pedreño García, María Ángeles; Biología Vegetal
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New strategies for the use of Linum usitatissimum cell factories for the production of bioactive compounds
(Elsevier, 2015-12-18) Almagro Romero, Lorena; Belchí Navarro, Sarai; Sánchez-Pujante, Pedro Joaquín; Pedreño García, María Ángeles; García Pérez, Pascual; Tecnología de Alimentos, Nutrición y Bromatología; Facultad de Veterinaria
In this work, suspension-cultured cells of Linum usitatissimum L. were used to evaluate the effect of two types of cyclodextrins, β-glucan and (Z)-3-hexenol separately or in combination on phytosterol and tocopherol production. Suspension-cultured cells of L. usitatissimum were able to produce high levels of phytosterols in the presence of 50 mM methylated-β-cyclodextrins (1325.96 ± 107.06 μg g dry weight−1) separately or in combination with β-glucan (1278.57 ± 190.10 μg g dry weight−1) or (Z)-3-hexenol (1507.88 ± 173.02 μg g dry weight−1), being cyclodextrins able to increase both the secretion and accumulation of phytosterols in the spent medium, whereas β-glucan and (Z)-3-hexenol themselves only increased its intracellular accumulation. Moreover, the phytosterol values found in the presence of hydroxypropylated-β-cyclodextrins were lower than those found in the presence of methylated-β-cyclodextrins in all cases studied. However, the results showed that the presence of methylated-β-cyclodextrins did not increase the tocopherols production and only an increase in tocopherol levels was observed when cells were elicited with 50 mM hydroxypropylated-β-cyclodextrins in combination with β-glucan (174 μg g dry weight−1) or (Z)-3-hexenol (257 μg g dry weight−1). Since the levels of tocopherol produced in the combined treatment were higher than the sum of the individual treatments, a synergistic effect between both elicitors was assumed. To sum up, flax cell cultures elicited with cyclodextrins alone or in combination with β-glucan or (Z)-3-hexenol were able produce phytosterols and tocopherols, and therefore, these elicited suspension-cultured cells of L. usitatissimum can provide an alternative system, which is at the same time more sustainable, economical and ecological for their production.
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Biotechnological approaches to enhance the biosynthesis of ginkgolides and bilobalide in Ginkgo biloba
(Springer, 2013-02-09) Souliman Youssef, Sabry; Novo Uzal, Esther; Almagro Romero, Lorena; Belchí Navarro, Sarai; Pedreño García, María Ángeles; Sabater Jara, Ana Belén; Biología Vegetal
Ginkgo biloba is one of the oldest living tree species and its extracts or powdered leaves are one of the best selling herbal preparations. The main bioactive constituents are flavonoids and the terpene trilactones, ginkgolides and bilobalide, which are responsible for their pharmacological activity. However, there are many difficulties for ginkgo leaves supply and the chemical synthesis is far from of being applicable for commercial-scale production. G. biloba cell cultures have arisen as a useful alternative source of pharmacologically active terpene trilactones. This review sheds light on the chemistry and biosynthesis of terpene trilactones with the aim of increasing the production of these high value compounds by biotechnological approaches. Different biotechnological strategies to improve ginkgolides and bilobalide production will be discussed, including screening and selection of in vitro ginkgo cultures, cell differentiation levels of these cultures, optimization of culture conditions, feeding and elicitation strategies. Special attention will be paid in developing new methodologies to enhance ginkgo cell biomass and provide high amounts of these bioactive terpene trilactones using large-scale cell cultures.