Fernando Prado - Academia.edu (original) (raw)

Papers by Fernando Prado

Research paper thumbnail of Cloning, structure and expression of a pea cDNA clone coding for a photosynthetic fructose-1,6-bisphosphatase with some features different from those of the leaf chloroplast enzyme

Planta, 1994

A positive clone against pea (Pisum sativum L.) chloroplast fructose-1,6-bisphosphatase (FBPase; ... more A positive clone against pea (Pisum sativum L.) chloroplast fructose-1,6-bisphosphatase (FBPase; EC 3.1.3.11) antibodies was obtained from a copy DNA (cDNA) library in lambda gt11. The insert was 1261 nucleotides long, and had an open reading frame of 1143 base pairs with coding capability for the whole FBPase subunit and a fragment of a putative processing peptide. An additional 115 base pairs corresponding to a 3'-untranslated region coding for an mRNA poly(A)+ tail were also found in the clone. The deduced sequence for the FBPase subunit was a 357-amino-acid protein of molecular mass 39,253 daltons (Da), showing 82-88% absolute homology with four chloroplastic FBPases sequenced earlier. The 3.1-kilobase (kb) KpnI-SacI fragment of the lambda gt11 derivative was subcloned between the KpnI-SacI restriction sites of pTZ18R to yield plasmid pAMC100. Lysates of Escherichia coli (pAMC100) showed FBPase activity; this was purified as a 170-kDa protein which, upon sodium dodecyl sulfate-polyacrylamide gel electrophoresis, displayed a 44-kDa band. As occurs with native FBPases, this indicates a homotetrameric structure for the expressed FBPase. When assayed under excess Mg2+ (10 mM), the expressed enzyme had a higher affinity for the substrate than the native pea leaf FBPase; this parameter appears to be substantiated by a tenfold higher specific activity than that of the native enzyme. However, when activated with dithiothreitol plus saturating concentrations of pea thioredoxin (Td) f, both FBPase had similar activities, with a 4:1 Td f-FBPase stoichiometry. In contrast to the native pea chloroplast FBPase, the E. coli-expressed enzyme did not react with the monoclonal antibody GR-PB5.(ABSTRACT TRUNCATED AT 250 WORDS)

Research paper thumbnail of Antigenic Relationships between Chloroplast and Cytosolic Fructose-1,6-Bisphosphatases

Plant Physiology, 1994

Cytosolic fructose-1,6-biphosphatases (FBPase, EC 3.1.3.1 1) from pea (Pisum safivum 1. cv Lincol... more Cytosolic fructose-1,6-biphosphatases (FBPase, EC 3.1.3.1 1) from pea (Pisum safivum 1. cv Lincoln) and spinach (Spinacia oleracea 1. cv Winter Ciant) did not cross-react by double immu

Research paper thumbnail of Cloning and Sequencing of a Pea cDNA Fragment Coding for Thioredoxin m

Research paper thumbnail of Seasonal variability of physiological and biochemical aspects of chromium accumulation in outdoor-grown Salvinia minima

Chemosphere, 2010

Seasonal variations in physiological and biochemical parameters of the aquatic fern Salvinia mini... more Seasonal variations in physiological and biochemical parameters of the aquatic fern Salvinia minima exposed to different Cr(VI) concentrations were studied. Growth, photosynthetic pigments, soluble carbohydrates, sucrose-related enzymes, lipid peroxidation, phenolics, and Cr accumulation in floating and submerged leaves were analyzed. Cr content was lower in winter than in summer, indicating that active metabolic events occurred in metal uptake. Leaf number and metal concentration factor were higher in summer than in winter. Relative growth rate (R n) indicated that growth was more affected by Cr in winter than in summer. Biochemical parameters showed great seasonal variations under increasing Cr. Hexose, starch, malondialdehyde and phenolic contents were greatest in winter, but R n and protein values were lowest. Sucrose content was highest in summer floating leaves. A great seasonal variability was observed in sucrose-related enzymes with the highest activities occurring in winter lipoxygenase was much higher in winter than in summer, indicating a strong lipid peroxidation. Results indicate that in Salvinia Cr causes seasonal perturbations in carbohydrate metabolism and oxidative stress by altering both sucrose-related enzymes and lipoxygenase activities. Variability in physiological and biochemical parameters seems to indicate that in outdoor conditions different mechanisms, in terms of Cr accumulation and tolerance, may occur in S. minima during summer and winter.

Research paper thumbnail of La fertirrigación con vinaza de caña de azúcar limita la tasa fotosintética de soja (Glycine max, Leguminosae)

Boletín de la Sociedad Argentina de Botánica

Introducción y Objetivos: En las provincias argentinas productoras de alcohol se genera anualment... more Introducción y Objetivos: En las provincias argentinas productoras de alcohol se genera anualmente un gran volumen de vinazas. Se ha propuesto que las mismas podrían ser utilizada para el fertirriego de la caña de azúcar u otras especies. Sin embargo, las investigaciones básicas para conocer el efecto de las mismas sobre el suelo, morfología y fisiología de las especies a fertirrigar son escasas en el noroeste argentino. M&M: Se aportan datos sobre el efecto de la vinaza diluida (1:1, 1:5 y 1:10 v/v) sobre la asimilación fotosintética de soja cultivada bajo condiciones semicontroladas (invernáculo). Resultados: La asimilación fotosintética máxima (Amax) se afecta fuertemente en todas las diluciones utilizadas. En la menor dilución (1:1) la Amax disminuye el 35 % respecto al control, mientras que en la dilución 1:10 dicho porcentaje se reduce al 15,9 %. Los cambios en la Amax se correlacionaron con disminuciones en la conductancia estomática, contenido de nitrógeno foliar (Nf) y clor...

Research paper thumbnail of Cloning, structure and expression of a pea cDNA clone coding for a photosynthetic fructose-1,6-bisphosphatase with some features different from those of the leaf chloroplast enzyme

Planta, 1994

A positive clone against pea (Pisum sativum L.)

Research paper thumbnail of Cloning, structure and expression of a pea cDNA clone coding for a photosynthetic fructose-1,6-bisphosphatase with some features different from those of the leaf chloroplast enzyme

Planta, 1994

A positive clone against pea (Pisum sativum L.) chloroplast fructose-1,6-bisphosphatase (FBPase; ... more A positive clone against pea (Pisum sativum L.) chloroplast fructose-1,6-bisphosphatase (FBPase; EC 3.1.3.11) antibodies was obtained from a copy DNA (cDNA) library in lambda gt11. The insert was 1261 nucleotides long, and had an open reading frame of 1143 base pairs with coding capability for the whole FBPase subunit and a fragment of a putative processing peptide. An additional 115 base pairs corresponding to a 3'-untranslated region coding for an mRNA poly(A)+ tail were also found in the clone. The deduced sequence for the FBPase subunit was a 357-amino-acid protein of molecular mass 39,253 daltons (Da), showing 82-88% absolute homology with four chloroplastic FBPases sequenced earlier. The 3.1-kilobase (kb) KpnI-SacI fragment of the lambda gt11 derivative was subcloned between the KpnI-SacI restriction sites of pTZ18R to yield plasmid pAMC100. Lysates of Escherichia coli (pAMC100) showed FBPase activity; this was purified as a 170-kDa protein which, upon sodium dodecyl sulfate-polyacrylamide gel electrophoresis, displayed a 44-kDa band. As occurs with native FBPases, this indicates a homotetrameric structure for the expressed FBPase. When assayed under excess Mg2+ (10 mM), the expressed enzyme had a higher affinity for the substrate than the native pea leaf FBPase; this parameter appears to be substantiated by a tenfold higher specific activity than that of the native enzyme. However, when activated with dithiothreitol plus saturating concentrations of pea thioredoxin (Td) f, both FBPase had similar activities, with a 4:1 Td f-FBPase stoichiometry. In contrast to the native pea chloroplast FBPase, the E. coli-expressed enzyme did not react with the monoclonal antibody GR-PB5.(ABSTRACT TRUNCATED AT 250 WORDS)

Research paper thumbnail of Antigenic Relationships between Chloroplast and Cytosolic Fructose-1,6-Bisphosphatases

Plant Physiology, 1994

Cytosolic fructose-1,6-biphosphatases (FBPase, EC 3.1.3.1 1) from pea (Pisum safivum 1. cv Lincol... more Cytosolic fructose-1,6-biphosphatases (FBPase, EC 3.1.3.1 1) from pea (Pisum safivum 1. cv Lincoln) and spinach (Spinacia oleracea 1. cv Winter Ciant) did not cross-react by double immu

Research paper thumbnail of Cloning and Sequencing of a Pea cDNA Fragment Coding for Thioredoxin m

Research paper thumbnail of Seasonal variability of physiological and biochemical aspects of chromium accumulation in outdoor-grown Salvinia minima

Chemosphere, 2010

Seasonal variations in physiological and biochemical parameters of the aquatic fern Salvinia mini... more Seasonal variations in physiological and biochemical parameters of the aquatic fern Salvinia minima exposed to different Cr(VI) concentrations were studied. Growth, photosynthetic pigments, soluble carbohydrates, sucrose-related enzymes, lipid peroxidation, phenolics, and Cr accumulation in floating and submerged leaves were analyzed. Cr content was lower in winter than in summer, indicating that active metabolic events occurred in metal uptake. Leaf number and metal concentration factor were higher in summer than in winter. Relative growth rate (R n) indicated that growth was more affected by Cr in winter than in summer. Biochemical parameters showed great seasonal variations under increasing Cr. Hexose, starch, malondialdehyde and phenolic contents were greatest in winter, but R n and protein values were lowest. Sucrose content was highest in summer floating leaves. A great seasonal variability was observed in sucrose-related enzymes with the highest activities occurring in winter lipoxygenase was much higher in winter than in summer, indicating a strong lipid peroxidation. Results indicate that in Salvinia Cr causes seasonal perturbations in carbohydrate metabolism and oxidative stress by altering both sucrose-related enzymes and lipoxygenase activities. Variability in physiological and biochemical parameters seems to indicate that in outdoor conditions different mechanisms, in terms of Cr accumulation and tolerance, may occur in S. minima during summer and winter.

Research paper thumbnail of La fertirrigación con vinaza de caña de azúcar limita la tasa fotosintética de soja (Glycine max, Leguminosae)

Boletín de la Sociedad Argentina de Botánica

Introducción y Objetivos: En las provincias argentinas productoras de alcohol se genera anualment... more Introducción y Objetivos: En las provincias argentinas productoras de alcohol se genera anualmente un gran volumen de vinazas. Se ha propuesto que las mismas podrían ser utilizada para el fertirriego de la caña de azúcar u otras especies. Sin embargo, las investigaciones básicas para conocer el efecto de las mismas sobre el suelo, morfología y fisiología de las especies a fertirrigar son escasas en el noroeste argentino. M&M: Se aportan datos sobre el efecto de la vinaza diluida (1:1, 1:5 y 1:10 v/v) sobre la asimilación fotosintética de soja cultivada bajo condiciones semicontroladas (invernáculo). Resultados: La asimilación fotosintética máxima (Amax) se afecta fuertemente en todas las diluciones utilizadas. En la menor dilución (1:1) la Amax disminuye el 35 % respecto al control, mientras que en la dilución 1:10 dicho porcentaje se reduce al 15,9 %. Los cambios en la Amax se correlacionaron con disminuciones en la conductancia estomática, contenido de nitrógeno foliar (Nf) y clor...

Research paper thumbnail of Cloning, structure and expression of a pea cDNA clone coding for a photosynthetic fructose-1,6-bisphosphatase with some features different from those of the leaf chloroplast enzyme

Planta, 1994

A positive clone against pea (Pisum sativum L.)