Luis del Río - Academia.edu (original) (raw)

Papers by Luis del Río

Biologia Plantarum, 2011

The role of peroxisomes in the oxidative injury induced by the auxin herbicide 2,4-dichlorophenox... more The role of peroxisomes in the oxidative injury induced by the auxin herbicide 2,4-dichlorophenoxyacetic acid (2,4-D) in leaves of pea (Pisum sativum L.) plants was studied. Applications of (2,4-D) on leaves or to root substrate increased the superoxide radical production in leaf peroxisomes. Foliar application also increased H 2 O 2 contents in leaf peroxisomes. Reactive oxygen species (ROS) overproduction was accompanied by oxidative stress, as shown by the changes in lipid peroxidation, protein carbonyls, total and protein thiols, and by the up-regulation of the activities of superoxide dismutase, ascorbate peroxidase, glutathione reductase, catalase, glucose 6-phosphate dehydrogenase and NADP +-dependent isocitrate dehydrogenase. Foliar or root 2,4-D applications also induced senescence symptoms in pea leaf peroxisomes, as shown by the decrease of protein content and glycolate oxidase and hydroxypyruvate reductase activities, and by the increase of endopeptidase, xanthine oxidase, isocitrate lyase and acyl-CoA oxidase activities as well as of 3-ketoacyl-CoA thiolase and thiol-protease protein contents. 2,4-D did not induce proliferation of pea leaf peroxisomes but induced senescence-like morphological changes in these organelles. Results suggest that peroxisomes might contribute to 2,4-D toxicity in pea leaves by overproducing cell-damaging ROS and by participating actively in 2,4-D-induced leaf senescence.

Nitric oxide ( • NO) is a gaseous free radical which can have two opposite physiological roles in... more Nitric oxide ( • NO) is a gaseous free radical which can have two opposite physiological roles in higher plants. While a high cellular production ofNO can bring about extensive cellular damage, at low levels this compound is involved as a signal molecule in many important physiological processes. • NO and a family of related molecules, including peroxynitrite (ONOO - ), S-nitrosoglutathione (GSNO) and nitrotyrosine (nTyr), among others, are designated with the term reactive nitrogen species (RNS). Under stress conditions, plants can undergo a de-regulated synthesis or overproduction ofNO and NO-derived products that can have toxic physiological consequences. This situation is known as nitrosative stress, due to its similarities with animal systems, and can produce important changes in plant cells. In this article the current knowledge of these effects of RNS on the physiology of plants under stress conditions is briefly reviewed.

Annals of Botany, 2015

Background and Aims Pepper (Capsicum annuum) contains high levels of antioxidants, such as vitami... more Background and Aims Pepper (Capsicum annuum) contains high levels of antioxidants, such as vitamins A and C and flavonoids. However, information on the role of these beneficial compounds in the physiology of pepper fruit remains scarce. Recent studies have shown that antioxidants in ripe pepper fruit play a key role in responses to temperature changes, and the redox state at the time of harvest affects the nutritional value for human consumption. In this paper, the role of antioxidant metabolism of pepper fruit during ripening and in the response to low temperature is addressed, paying particular attention to ascorbate, NADPH and the superoxide dismutase enzymatic system. The participation of chloroplasts, mitochondria and peroxisomes in the ripening process is also investigated. Scope and Results Important changes occur at a subcellular level during ripening of pepper fruit. Chloroplasts turn into chromoplasts, with drastic conversion of their metabolism, and the role of the ascorbate-glutathione cycle is essential. In mitochondria from red fruits, higher ascorbate peroxidase (APX) and Mn-SOD activities are involved in avoiding the accumulation of reactive oxygen species in these organelles during ripening. Peroxisomes, whose antioxidant capacity at fruit ripening is substantially affected, display an atypical metabolic pattern during this physiological stage. In spite of these differences observed in the antioxidative metabolism of mitochondria and peroxisomes, proteomic analysis of these organelles, carried out by 2-D electrophoresis and MALDI-TOF/TOF and provided here for the first time, reveals no changes between the antioxidant metabolism from immature (green) and ripe (red) fruits. Conclusions Taken together, the results show that investigation of molecular and enzymatic antioxidants from cell compartments, especially chloroplasts, mitochondria and peroxisomes, is a useful tool to study the physiology of pepper fruit, particularly in the context of expanding their shelf-life after harvest and in maintaining their nutritional value.

SOD) located in the matrix of peroxisomes from watermelon (Citrullus vulgaris Schrad.) cotyledons... more SOD) located in the matrix of peroxisomes from watermelon (Citrullus vulgaris Schrad.) cotyledons (L.M. Sandalio and L.A. del Rio [1988] Plant Physiol 88: 1215-1218). The amino acid composition of the enzyme was determined. Analysis by reversed-phase high-performance liquid chromatography of the peroxisomal CuZnSOD incubated with 6 M guanidine-HCI indicated that this enzyme contained a noncovalently bound chromophore group that was responsible for the absorbance peak of the native enzyme at 260 nm. The amino acid sequence of the peroxisomal CuZn-SOD was determined by Edman degradation. Comparison of its sequence with those reported for other plant SODs revealed homologies of about 70% with cytosolic CuZn-SODs and of 90% with chloroplastic CuZn-SODs. The peroxisomal SOD has a high thermal stability

Horticultura Revista De Industria Distribucion Y Socioeconomia Horticola Frutas Hortalizas Flores Plantas Arboles Ornamentales Y Viveros, 2007

Nitric Oxide in Plant Physiology

... Francisco J. Corpas, José M. Palma, Marina Leterrier, Luis A. del Rıo, and Juan B. Barroso Su... more ... Francisco J. Corpas, José M. Palma, Marina Leterrier, Luis A. del Rıo, and Juan B. Barroso Summary ... 76 Corpas, FJ, Chaki, M., Fernández-Ocaña, A., Valderrama, R., Palma, JM, Carreras, A., Begara-Morales, JC, Airaki, M., del Rıo, LA, and Barroso, JB (2008)Plant Cell Physiol. ...

Plant Tolerance to Abiotic Stresses in Agriculture: Role of Genetic Engineering, 2000

In this work the effect of growing pea (Pisum sativum L.) plants with a toxic CdC12 concentration... more In this work the effect of growing pea (Pisum sativum L.) plants with a toxic CdC12 concentration on the activated oxygen metabolism of leaf peroxisomes was studied. Pea plants were grown in the greenhouse under optimum conditions for 14 days, and then media were supplemented with 50 µM CdC12 and grown for 21 days. Peroxisomes were purified from pea leaves

Planta, 1980

A manganese-containing superoxide dismutase (EC 1.15.1.1) was purified to homogeneity from a high... more A manganese-containing superoxide dismutase (EC 1.15.1.1) was purified to homogeneity from a higher plant for the first time. The enzyme was isolated fromPisum sativum leaf extracts by thermal fractionation, ammonium sulfate salting out, ion-exchange and gel-filtration column chromatography, and preparative polyacrylamide gel electrophoresis. Pure manganese superoxide dismutase had a specific activity of about 3,000 U mg(-1) and was purified 215-fold, with a yield of 1.2 mg enzyme per kg whole leaf. The manganese superoxide dismutase had a molecular weight of 94,000 and contained one g-atom of Mn per mol of enzyme. No iron and copper were detected. Activity reconstitution experiments with the pure enzyme ruled out the possibility of a manganese loss during the purification procedure. The stability of manganese superoxide dismutase at-20°C, 4°C, 25°C, 50°C, and 60°C was studied, and the enzyme was found more labile at high temperatures than bacterial manganese superoxide dismutases and iron superoxide dismutases from an algal and bacterial origin.

Planta, 1978

The effect of different Mn levels on the isozyme pattern of superoxide dismutase was investigated... more The effect of different Mn levels on the isozyme pattern of superoxide dismutase was investigated. Pisum sativum L. plants were grown in nutrient solutions containing three Mn concentrations: 0.005 μg/ml (deficient), 0.05 μg/ml (low), and 0.5 μg/ml (optimum). Leaf extracts contained three electrophoretically distinct superoxide dismutases (SOD), two of which were inhibited by cyanide and were probably Cu-Zn-SODs, while the third one was CN-insensitive and could be either an Mn- or an Fe-SOD. At 0.005 μg/ml Mn supply the CN-insensitive SOD was significantly depressed at 15, 30, and 45 days of growth, whereas at 0.05 μg/ml Mn this isozyme was significantly decreased only at 45 days growth. The two CN-sensitive SODs were inversely related to the CN-resistant enzyme, the activities of the former enzymes being significantly increased at Mn-deficient levels throughout plant growth. Metal determinations of the plants showed that at low concentrations of Mn in the nutrient media, copper and zinc content of leaves increased: the lower the Mn level, the higher the increase produced. The CN-resistant SOD activity, as judged by its dependency on Mn, appears to be an Mn-SOD rather than an Fe-SOD. In the light of the results obtained, the use of the enzyme system superoxide dismutase for the study of the role and interactions between Mn, Cu, and Zn in the plant cell is proposed.

Plant Physiology and Biochemistry, 2002

... and plants which enters the environment mainly from industrial processes and phosphate fertil... more ... and plants which enters the environment mainly from industrial processes and phosphate fertilizers and then is ... Cadmium effect on growth and physiological parameters of pepper. ... ions in the nutrient solution produced a significant growth inhibition of pepper plants, measured as ...

New Phytologist, 1994

Page 1. Л' Phyto!. (I"44). 126, .17-44 Salt stress-induced changes in superoxide dismu... more Page 1. Л' Phyto!. (I"44). 126, .17-44 Salt stress-induced changes in superoxide dismutase isozymes in leaves and mesophyll protoplasts from Vigna unguiculata (L.) Walp. By JOSÉ A. HERNANDEZ1. LUIS A. DEL RIO* and ...

Journal of Proteomics, 2011

Fruit ripening is a developmental complex process which occurs in higher plants and involves a nu... more Fruit ripening is a developmental complex process which occurs in higher plants and involves a number of stages displayed from immature to mature fruits that depend on the plant species and the environmental conditions. Nowadays, the importance of fruit ripening comes mainly from the link between this physiological process in plants and the economic repercussions as a result of one of the human activities, the agricultural industry. In most cases, fruit ripening is accompanied by colour changes due to different pigment content and increases in sugar levels, among others. Major physiological modifications that affect colour, texture, flavour, and aroma are under the control of both external (light and temperature) and internal (developmental gene regulation and hormonal control) factors. Due to the huge amount of metabolic changes that take place during ripening in fruits from higher plants, the accomplishment of new throughput methods which can provide a global evaluation of this process would be desirable. Differential proteomics of immature and mature fruits would be a useful tool to gain information on the molecular changes which occur during ripening, but also the investigation of fruits at different ripening stages will provide a dynamic picture of the whole transformation of fruits. This subject is furthermore of great interest as many fruits are essential for human nutrition. Thus far different maturation profiles have been reported specific for each crop species. In this work, a thorough review of the proteomic database from fruit development and maturation of important crop species will be updated to understand the molecular physiology of fruits at ripening stages.

Journal of Plant Physiology, 2003

Pepper is a vegetable of importance in human nutrition. Currently, one of the most interesting pr... more Pepper is a vegetable of importance in human nutrition. Currently, one of the most interesting properties of natural products is their antioxidant content. In this work, the purification and characterisation of peroxisomes from fruits of a higher plant was carried out, and their antioxidative enzymatic and non-enzymatic content was investigated. Green and red pepper fruits (Capsicum annuum L., type Lamuyo) were used in this study. The analysis by electron microscopy showed that peroxisomes from both types of fruits contained crystalline cores which varied in shape and size, and the presence of chloroplasts and chromoplasts in green and red pepper fruits, respectively, was confirmed. Peroxisomes were purified by differential and sucrose density-gradient centrifugations. In the peroxisomal fractions, the activity of the photorespiration, β-oxidation and glyoxylate cycle enzymes, and the ROS-related enzymes catalase, superoxide dismutase, xanthine oxidase, glutathione reductase and NADP +-dehydrogenases, was determined. Most enzymes studied had higher specific activity and protein content in green than in red fruits. By native PAGE and western blot analysis, the localisation of a Mn-SOD in fruit peroxisomes was demonstrated. The ascorbate and glutathione levels were also determined in crude extracts and in peroxisomes purified from both green and red peppers. The total ascorbate content (200-220 mg per 100 g FW) was similar in crude extracts from the two types of fruits, but higher in peroxisomes from red peppers. The glutathione concentration was 2-fold greater in green pepper crude extracts than in red fruits, whereas peroxisomes from both tissues showed similar values. The presence in pepper peroxisomes of different antioxidative enzymes and their corresponding metabolites implies that these organelles might be an important pool of antioxidants in fruit cells, where these enzymes could also act as modulators of signal molecules (O 2˙-, H 2 O 2) during fruit maturation.

Journal of Plant Physiology, 1993

Summary The effect of NaCl salinity on the metabolism of activated oxygen was studied in peroxiso... more Summary The effect of NaCl salinity on the metabolism of activated oxygen was studied in peroxisomes purified from leaves of two cultivars of Pisum sativum L. with different sensitivity to NaCl. Salinity caused a decrease in urate oxidase and hydroxypyruvate reductase activities in peroxisomes from NaCl-sensitive plants, whereas the activity of glycollate oxidase was stimulated by NaCl in salt-tolerant plants. Catalase activity was decreased by NaCl in both cultivars of Pisum sativum L. Xanthine oxidase was the most abundant form of xanthine oxidoreductases in pea leaf peroxisomes. Both xanthine oxidase and xanthine dehydrogenase were differentially influenced by NaCl depending on the pea cultivar, but there was no concomitant increase in the activity of xanthine oxidase, indicating that salinity does not affect the interconversion of xanthine dehydrogenase into xanthine oxidase. In peroxisomal membranes, neither NADH-dependent generation of O 2 - nor lipid peroxidation was altered by salinity. However, the endogenous concentration of H 2 O 2 in leaf peroxisomes was decreased by NaCl in both salt-tolerant and salt-sensitive plants. Thus, in NaCl-tolerance of Pisum sativum L. both the photorespiratory glycollate pathway and the urate oxidase-catalyzed biosynthesis of allantoin appear to be involved at the peroxisomal level.

Journal of Plant Nutrition, 1987

The evaluation of nutritional status of plants through the use of biological indicators has been ... more The evaluation of nutritional status of plants through the use of biological indicators has been found useful when chemical analysis of the leaf nutrient concentrations does not reveal certain nutrient imbalances. In this work, the responses of Superoxide dismutases (SOD) isoenzymes, chlorophylls and photosystem II activity of leaves from soybean plants grown under different Fe and Mn nutrient levels were

Journal of Inorganic Biochemistry, 1982

Homogenous preparations of a manganese superoxide dismutase from a higher plant (Pisum sarivunz L... more Homogenous preparations of a manganese superoxide dismutase from a higher plant (Pisum sarivunz L.) were studied by epr and optical spectroscopies. The visible spectrum of manganese superoxide dismutase shows a weak and broad band in the range 350-700 nm with two shouIden at about 480 and 600 nm. Reduction with dirhionite brought about a considerable disappearance of the visible component of the spectrum_ The epr spectra of the native and dithionite-treated enzyme did not show any signal attributable to Mn(I1) that only was visible after acid hydrolysis of rhe protein. The lack of epr signal both in the native and reduced superoxide dismutase can be attributed to the presence of Mn(II1) in the former and of Mn(I1) strongly bound to the protein in the latter. The results obtained with the manganese superoxide dismutase from leaves of the higher plant Pisum sativum are consistent with the general catalytic mechanism of action postulated for superoxide dismutases from other sources studied so far.

Free Radical Research Communications, 1991

A cyanide-insensitive superoxide dismutase was purified to apparent homogeneity from lemon leaves... more A cyanide-insensitive superoxide dismutase was purified to apparent homogeneity from lemon leaves (Citrus limonum R). The enzyme was isolated from leaf extracts by ammonium sulfate salting-out, and ion-exchange, gel filtration and hydroxylapatite column chromatography. The purified Fe-SOD had a specific activity of about 1,500 U/mg and represents approximately 1.6% of the total soluble protein in lemon leaf extracts. A molecular weight of 47,500 was determined for the enzyme. Analytical gel electro-focusing of the purified preparation revealed the presence of two isozymes with pI values of 5.13 and 4.98. Metal analysis showed the presence of 1 g-atom of iron and 0.5 g-atom of manganese per mol of enzyme. The visible and UV absorption spectra of the Citrus enzyme were similar to those reported for other iron-containing SODs from different origins. The significance of the presence of Fe-SOD in higher plants is briefly discussed.

Acta Physiologiae Plantarum, 2011

Oxidative stress and senescence have been shown to participate in the toxicity mechanism of auxin... more Oxidative stress and senescence have been shown to participate in the toxicity mechanism of auxin herbicides in the leaves and roots of sensitive plants. However, their role in stem toxicity has not been studied yet. In this work, we report the effect of foliar or root applications of the auxin herbicide 2,4-dichlorophenoxyacetic acid (2,4-D) on the parameters of oxidative stress and senescence of stems of pea (Pisum sativum L.) plants. Contrary to their effect on the pea leaves, in the stems 2,4-D applications did not cause oxidative stress, as shown by the parameters of lipid peroxidation, protein carbonyls, and total and protein thiols. Moreover, they inhibited the superoxide radical (O 2 .-)-producing xanthine oxidase (XOD) activity and stimulated the antioxidant activities of catalase (CAT), guaiacol peroxidase (GPOX), ascorbate peroxidase (APX), glutathione reductase (GR), glutathione S-transferase (GST) and Krebs cycle NAD ?-isocitrate dehydrogenase (IDH). Applications of 2,4-D also did not induce senescence in the pea stems, as shown by the increase of proteins, the lack of stimulation of proteolytic activity, and the inhibition of senescence-related isocitrate lyase (ICL) activity. However, they stimulated the H 2 O 2-producing acyl-CoA oxidase (ACOX) activity of fatty acid beta oxidation. Results suggest that oxidative stress and senescence are not involved in the mechanism of toxicity of 2,4-D in the stems of pea plants, and that these phenomena are not whole-plant toxicity mechanisms for auxin herbicides in susceptible plants. Results also suggest that the effect of 2,4-D on the oxidative metabolism of pea plants might be organ-specific. Keywords Auxin herbicides Á 2,4-Dichlorophenoxyacetic acid Á Oxidative stress Á Pea Á Senescence Á Stems Abbreviations AA Ascorbic acid ACOX Acyl-CoA oxidase APX Ascorbate peroxidase CAT Catalase C=O Protein carbonyls F4d 4-day foliar treatment R4d 4-day root treatment R7d 7-day root treatment

Journal of Integrative Plant Biology, 2019

Plant peroxisomes are subcellular compartments involved in many biochemical pathways during the l... more Plant peroxisomes are subcellular compartments involved in many biochemical pathways during the life cycle of a plant but also in the mechanism of response against adverse environmental conditions. These organelles have an active nitro-oxidative metabolism under physiological conditions but this could be exacerbated under stress situations. Furthermore, peroxisomes have the capacity to proliferateand also undergo biochemical adaptations depending on the surrounding cellular status. An important characteristic of peroxisomes is that they have a dynamic metabolism of reactive nitrogen and oxygen species (RNS and ROS) which generates two key molecules, nitric oxide (NO) and hydrogen peroxide (H 2 O 2). These molecules can exert signaling functions by means of post-translational modifications that affect the functionality of target molecules like proteins, peptides or fatty acids. This review provides an overview of the endogenous metabolism of ROS and RNS in peroxisomes with special emphasis on polyamine and uric acid metabolism as well as the possibility that these organelles could be a source of signal molecules involved in the functional interconnection with other subcellular compartments.

Acta Physiologiae Plantarum, 2017

Peroxiredoxins (Prxs) constitute a group of thiolspecific antioxidant enzymes which are present i... more Peroxiredoxins (Prxs) constitute a group of thiolspecific antioxidant enzymes which are present in bacteria, yeasts, and in plant and animal cells. Although Prxs are mainly localized in the cytosol, they are also present in mitochondria, chloroplasts, and nuclei, but there is no evidence of the existence of Prxs in plant peroxisomes. Using soluble fractions (matrices) of peroxisomes purified from leaves of pea (Pisum sativum L.) plants, the immunological analysis with affinity-purified IgG against yeast Prx1 revealed the presence of an immunoreactive band of about 50 kDa. The apparent molecular mass of the peroxisomal Prx was not sensitive to oxidizing and reducing conditions what could be a mechanism of protection against the oxidative environment existing in peroxisomes. Postembedment, EM immunocytochemical analysis with affinity-purified IgG against yeast Prx1 antibodies, confirmed that this protein was present in the peroxisomal matrix, mitochondria, and chloroplasts. In pea plants grown under oxidative stress conditions, the protein level of peroxisomal Prx was differentially modulated, being slightly induced by growth of plants with 50 lM CdCl 2 , but being significantly reduced by treatment with the herbicide 2,4-dichlorophenoxyacetic acid (2,4-D). The presence in the matrix of peroxisomes of a protein immunorelated to Prx of about 50 kDa, which is in the range of molecular mass of the dimeric form of other Prxs, opens new questions on the molecular properties of Prxs, but also on their function in the metabolism of reactive oxygen and nitrogen species (ROS/RNS) in these plant cell organelles, where they could be involved in the regulation of hydrogen peroxide and/or peroxynitrite.