Showing papers by "University of Córdoba (Spain) published in 2001"

A MAP kinase of the vascular wilt fungus Fusarium oxysporum is essential for root penetration and pathogenesis

Abstract: Summary The soil-borne vascular wilt fungus Fusarium oxysporum infects a wide variety of plant species by directly penetrating roots, invading the cortex and colonizing the vascular tissue. We have identified fmk1, encoding a mitogen-activated protein kinase (MAPK) of F. oxysporum that belongs to the yeast and fungal extracellular signal-regulated kinase (YERK1) subfamily. Targeted mutants of F. oxysporum f. sp. lycopersici carrying an inactivated copy of fmk1 have lost pathogenicity on tomato plants but show normal vegetative growth and conidiation in culture. Colonies of the fmk1 mutants are easily wettable, and hyphae are impaired in breaching the liquid‐air interface, suggesting defects in surface hydrophobicity. Fmk1 mutants also show reduced invasive growth on tomato fruit tissue and drastically reduced transcript levels of pl1 encoding the cell wall-degrading enzyme pectate lyase. Conidia of the mutants germinating in the tomato rhizosphere fail to differentiate penetration hyphae, resulting in greatly impaired root attachment. The orthologous MAPK gene Pmk1 from the rice leaf pathogen Magnaporthe grisea complements invasive growth and partially restores surface hydrophobicity, root attachment and pathogenicity in an fmk1 mutant. These results demonstrate that FMK1 controls several key steps in the pathogenesis of F. oxysporum and suggest a fundamentally conserved role for the corresponding MAPK pathway in soil-borne and foliar plant pathogens.

Hydrogenolysis of organohalogen compounds over palladium supported catalysts

Abstract: Catalytic hydrodehalogenation is one of the most promising methods for detoxification of organohalogen waste. A variety of catalytic systems has been used, although palladium catalysts are the best choice because of their optimal catalytic properties. Catalytic hydrodehalogenation on palladium catalysts is carried out in both the liquid and gas-phases. The experimental procedures for the catalytic hydrodehalogenation are reviewed; the hydrogen source can be either molecular hydrogen or a hydrogen donor in the so called hydrogen-transfer hydrogenolysis. A wide variety of hydrogen donors has been used, including inorganic salts (phosphinates) organic salts (formates), alcohols, proaromatic compounds, etc. Kinetics and mechanisms proposed for hydrodehalogenation are presented, and both agreements and disagreements on the mechanistic aspects are discussed. Catalysts modification (base addition, support modification, second metal addition, etc.) as well as its transformation during the hydrogenolytic process (i.e. deactivation) are discussed.

A Mediterranean and a high-carbohydrate diet improve glucose metabolism in healthy young persons

Abstract: Aims/hypothesis: Insulin resistance usually precedes the diagnosis of Type II (non-insulin-dependent) diabetes mellitus. However, in most patients, the clinical expression of the disease could be prevented by dietary and lifestyle changes. We investigated the effects of a diet enriched in monounsaturated fatty acids (Mediterranean diet) and a low fat, high-carbohydrate diet on in vivo and in vitro glucose metabolism in 59 young subjects (30 men and 29 women).

Comparison of leaf construction costs in woody species with differing leaf life-spans in contrasting ecosystems

Abstract: ummary • The construction costs (CC) are reported of leaves from 162 wild woody species from 14 contrasting environments (desert to rain forest) and with different leaf life-spans. • Calorimetric methods were used to estimate the CC of deciduous, semideciduous and evergreen leaves. • Leaf CC showed a wide range (78%) between species, and deciduous species showed a slightly lower CC (6%) than both semideciduous and evergreen species. Mean leaf CC differed between ecosystems, with the highest and lowest CC in the tundra and rain forest, respectively. Leaf CC was positively correlated with lipid concentration. Leaf size (log) and specific leaf area (SLA, leaf area per leaf dry mass) were negatively correlated with leaf CC. Leaf CC did not show differences between different leaf life-spans or ecosystems when leaf size (log) or SLA were included as covariates. • The small differences in leaf CC among leaf life-span types and ecosystems (6% and 23%, respectively) suggest that SLA is more important in determining differences in the carbon balance between species than leaf CC. Leaf size is shown to be an important trait associated with other leaf characteristics. Abbreviations A, ash concentration; CC, construction cost per unit dry mass; CCA, construction cost per unit area; Eg, growth efficiency; Hc, ash free heat of combustion; N, nitrogen; SLA, specific leaf area.

Sunflower (Helianthus annuus L.) response to broomrape (Orobanche cernua Loefl.) parasitism: induced synthesis and excretion of 7‐hydroxylated simple coumarins

Abstract: The interaction of the parasitic plant Orobanche cernua with resistant and susceptible cultivars of Helianthus annuus L. was investigated. Using different bioassays to evaluate the early stages of the parasite life cycle (germination, attachment, penetration, and establishment), differences were observed between O. cernua-resistant and O. cernua-susceptible sunflower varieties. Germination of O. cernua seeds in the presence of resistant sunflower roots was approximately half that of germination in the presence of susceptible roots, and germinated seeds displayed enhanced browning symptoms. Parasite radicles or host-tissue around the contact point turned brown after O. cernua attachment to sunflower roots, especially in the resistant varieties. These observations suggested the possible accumulation of toxic compounds as a defence strategy in the resistant sunflower varieties. Sunflower 7-hydroxylated simple coumarins may play a defensive role against O. cernua parasitism by preventing successful germination, penetration and/or connection to the host vascular system. This hypothesis is supported by the following data: (i) coumarins inhibited the in vitro germination of O. cernua seeds induced by the strigol analogue GR(24) and caused a browning reaction in germinated seeds and (ii) resistant sunflowers accumulated higher levels of coumarins in roots and excreted greater amounts than susceptible varieties in response to O. cernua infection.

Relative growth rate in phylogenetically related deciduous and evergreen woody species

Abstract: Relative growth rate (RGR) and other growth parameters were studied in eight pairs of closely related deciduous and evergreen species (within the same genus or family). The main objective of this study was to test the association between leaf turnover rate and RGR, specific leaf area (SLA, leaf area/leaf dry weight) and other growth variables. Plants were grown for 6 months in a greenhouse under favourable water and nutrient conditions. Variation in RGR among the 16 woody species was due mainly to differences in morphological parameters such as leaf area ratio (LAR, whole plant area/whole plant dry weight) and SLA). However, temporal variation in RGR within species was due mainly to variation in net assimilation rate. When phylogeny was not taken into account, analyses showed that deciduous species grew faster than evergreens. In contrast, when phylogeny was taken into account, the data analysis showed that a faster RGR is not consistently associated with the deciduous habit (in five pairs it was, but in the other three it was not). The faster growth of the deciduous trees (in the five positive contrasts) could be explained by their higher LAR and higher SLA relative to evergreens. The lack of differences in RGR between deciduous and evergreens (in three pairs) was due to the higher leaf mass ratio (LMR, leaf dry biomass/total dry biomass) for the evergreens, which offset the higher SLA of the deciduous species, resulting in a similar LAR in both functional groups (LAR=LMR×SLA). Deciduous species had consistently higher SLA than evergreens. We suggest that SLA, more than RGR, could be an important parameter in determining adaptive advantages of deciduous and evergreen species.

Molecular mechanisms of leptin action in adult rat testis: potential targets for leptin-induced inhibition of steroidogenesis and pattern of leptin receptor messenger ribonucleic acid expression

Abstract: Leptin, the product of the ob gene, is a pivotal signal in the regulation of neuroendocrine function and fertility. Although much of the action of leptin in the control of the reproductive axis is exerted at the hypothalamic level, some direct effects of leptin on male and female gonads have also been reported. Indeed, recent evidence demonstrated that leptin is able to inhibit testosterone secretion at the testicular level. However, the molecular mechanisms behind this effect remain unclear. The focus of this study was twofold: (1) to identify potential targets for leptin-induced inhibition of steroidogenesis, and (2) to characterize in detail the pattern of expression and cellular distribution of leptin receptor (Ob-R) mRNA in adult rat testis. In pursuit of the first goal, slices of testicular tissue from adult rats were incubated with increasing concentrations of recombinant leptin (10(-9)--10(-7 )M) in the presence of human chorionic gonadotropin (hCG; 10 IU/ml). In this setting, testosterone secretion in vitro was monitored, and expression levels of mRNAs encoding steroidogenic factor 1 (SF-1), steroidogenic acute regulatory protein (StAR), cytochrome P450 cholesterol side-chain cleavage enzyme (P450 scc) and 17 beta-hydroxysteroid dehydrogenase type III (17 beta-HSD) were assessed by Northern hybridization. In pursuit of the second goal, the pattern of cellular expression of the Ob-R gene in adult rat testis was evaluated by in situ hybridization using a riboprobe complementary to all Ob-R isoforms. In addition, testicular expression levels of the different Ob-R isoforms, previously identified in the hypothalamus, were analyzed by means of semi-quantitative RT-PCR. In keeping with our previous data, recombinant leptin significantly inhibited hCG-stimulated testosterone secretion. In this context, leptin, in a dose-dependent manner, was able to co-ordinately decrease the hCG-stimulated expression levels of SF-1, StAR and P450 scc mRNAs, but it did not affect those of 17 beta-HSD type III. In situ hybridization analysis showed a scattered pattern of cellular expression of the Ob-R gene within the adult rat testis, including Leydig and Sertoli cells. In addition, assessment of the pattern of expression of Ob-R subtypes revealed that the long Ob-Rb isoform was abundantly expressed in adult rat testis. However, variable levels of expression of Ob-Ra, Ob-Re, and Ob-Rf mRNAs were also detected, whereas those of the Ob-Rc variant were nearly negligible. In conclusion, our results indicate that decreased expression of mRNAs encoding several up-stream elements in the steroidogenic pathway may contribute, at least partially, to leptin-induced inhibition of testicular steroidogenesis. In addition, our data on the pattern of testicular expression of Ob-R isoforms and cellular distribution of Ob-R mRNA may help to further elucidate the molecular mechanisms of leptin action in rat testis.

Eukaryotic nitrate and nitrite transporters.

Abstract: Nitrate transport is the key step controlling the amount of nitrate incorporated by the cells and subsequent of storage, reduction or export. Molecular, genetic and biochemical approaches to the study of eukaryotic nitrate/nitrite transporters allow an initial understanding of this step, which is much more complex and structured than previously suspected. At the plasma membrane level, two gene families, Nrt1 and Nrt2, account for high- and low-affinity nitrate transporters. Functionality of NRT1 from Arabidopsis and NRT2 proteins from Aspergillus and Chlamydomonas has been demonstrated. However, redundancy of these systems makes it difficult to assign particular physiological roles to each. Data on genes involved in the regulation of nitrate transport and reduction are still scarce. Information on nitrite transporters to the chloroplast is biased by the belief that in vivo nitrous acid diffuses freely to this organellum. The recent progress on these aspects is discussed in this review.

Model for forecasting Olea europaea L. airborne pollen in South-West Andalusia, Spain.

Abstract: Data on predicted average and maximum airborne pollen concentrations and the dates on which these maximum values are expected are of undoubted value to allergists and allergy sufferers, as well as to agronomists. This paper reports on the development of predictive models for calculating total annual pollen output, on the basis of pollen and weather data compiled over the last 19 years (1982–2000) for Cordoba (Spain). Models were tested in order to predict the 2000 pollen season; in addition, and in view of the heavy rainfall recorded in spring 2000, the 1982–1998 data set was used to test the model for 1999. The results of the multiple regression analysis show that the variables exerting the greatest influence on the pollen index were rainfall in March and temperatures over the months prior to the flowering period. For prediction of maximum values and dates on which these values might be expected, the start of the pollen season was used as an additional independent variable. Temperature proved the best variable for this prediction. Results improved when the 5-day moving average was taken into account. Testing of the predictive model for 1999 and 2000 yielded fairly similar results. In both cases, the difference between expected and observed pollen data was no greater than 10%. However, significant differences were recorded between forecast and expected maximum and minimum values, owing to the influence of rainfall during the flowering period.

Parton densities and dipole cross-sections at small x in large nuclei

Abstract: Unintegrated gluon densities in nuclei, dipole-nucleus cross-sections and quark densities are numerically investigated in the high-color limit, with the scattering on a heavy nucleus exactly described by the sum of fan diagrams of BFKL pomerons. The initial condition for the evolution in rapidity is quickly forgotten, and the gluon density presents a “supersaturation” pattern, as previous studies indicated. Both dipole-nucleus cross-sections and quark densities present the expected saturation features. Identifying the position in transverse momentum l of the maximum of the gluon distribution with the saturation momentum $Q_{\mathrm{s}}(y,b)$ , at large rapidities all distributions depend on only one variable, $l/Q_{\mathrm{s}}(y,b)$ or $rQ_{\mathrm{s}}(y,b)$ .

Assessment of mechanisms of thyroid hormone action in mouse Leydig cells: regulation of the steroidogenic acute regulatory protein, steroidogenesis, and luteinizing hormone receptor function.

Abstract: Recently, we demonstrated that triiodothyronine (T3) stimulated steroid hormone biosynthesis and steroidogenic acute regulatory (StAR) protein expression in mLTC-1 mouse Leydig tumor cells through the mediation of steroidogenic factor 1 (SF-1). We now report a dual response mechanism of T3 on steroidogenesis and StAR expression, and on LH receptor (LHR) expression and binding in mLTC-1 cells. T3 acutely (8 h), induced a 260% increase in StAR messenger RNA (mRNA) expression over the basal level which was coincident with an increase in progesterone (P) production. In contrast, chronic stimulation with T3 (beyond 8 h), resulted in an attenuation of StAR expression and P production. This attenuation was most likely caused by a decrease in cholesterol delivery to the inner mitochondrial membrane as demonstrated by incubations with the hydrophilic steroid precursors, 22R hydroxycholesterol and pregnenolone, which restored P synthesis. In similar studies, chronic treatment with T3 increased the levels of cytochr...

Study of the degradation of the herbicides 2,4-D and MCPA at different depths in contaminated agricultural soil.

Abstract: Two phenoxyacid herbicides (2,4-D and MCPA) and their six corresponding phenols were determined in soil by using gas chomatography with electron impact mass spectrometry (GC/MS) for confirmation/quantitation. An automatic extraction (leaching), preconcentration, and cleanup (sorption) module was developed to extract the eight compounds from soil. The average recovery of all species, spiked to soil at microg/kg-mg/kg levels, was 95% (average standard deviation +/- 5%). A plot of agricultural clayey soil (approximately 12 m2) was contaminated with both herbicides (approximately 96 g/m3, depth 10 cm, density 1.23 g/cm3) and irrigated with (17 mm) at variable time intervals. Both herbicides and their corresponding phenol compounds were monitored at different soil depths over a 50 day period. The degradation of both herbicides in the surface layer (t(1/2) approximately 5 days) is a result of photodecomposition and microbial action; in the deeper layers, the degradation products occur in lower proportions by effect of leaching and are also the result of microbial action. The six phenol metabolites are only detected in the surface layer as they form preferentially by photodecomposition. The main metabolites (viz. 2,4-DCP for 2,4-D and 4-C-2-MP for MCPA) are formed within 24 h after the soil is contaminated; their concentration peaks are at day 8 in the absence of irrigation.

Developmental and Hormonal Regulation of Leptin Receptor (Ob-R) Messenger Ribonucleic Acid Expression in Rat Testis

Abstract: In target tissues, leptin receptor (Ob-R) gene expression results in an array of alternatively spliced isoforms (Ob-Ra to ObRf) with different functional features. Recent evidence has pointed to a direct role of leptin in the control of testicular function. However, complete elucidation of the pattern of Ob-R gene expression in the male gonad is still pending. The focus of this study was to characterize in detail the developmental pattern of expression and hormonal regulation of Ob-R gene in rat testis. To this end, the overall expression of Ob-R mRNA was compared to that of the fully functional, long Ob-Rb isoform in different experimental settings, using semiquantitative reverse transcription-polymerase chain reaction. Expression of Ob-R mRNA was detected in testes from 15-, 30-, 45-, and 75-day-old rats at rather constant relative levels. In contrast, testicular expression of Ob-Rb mRNA was higher in pubertal testes (15- to 30-day-old rats) and declined in adulthood. In testes from 30-day-old animals, analysis of isoform distribution revealed that, in addition to abundant Ob-Rb mRNA levels, expression of Ob-Ra, Ob-Rf, and, to a lesser extent, Ob-Rc and Ob-Re messages is detected. Testicular Ob-R mRNA expression appeared sensitive to neonatal imprinting as neonatal treatment with estradiol benzoate (500 mg/rat; Day 1 postpartum) resulted in a persistent increase (P , 0.01) in the relative expression level of Ob-R mRNA, a phenomenon only partially mimicked by neonatal suppression of serum gonadotropins by means of LHRH-antagonist administration. In addition, neonatal estrogenization differentially altered the pattern of expression of Ob-R isoforms in adult rat testis, as expression of Ob-Rb mRNA was decreased to undetectable levels, whereas that of Ob-Rc remained unaltered, and Ob-Ra, Ob-Rf, and, to a lesser extent, Ob-Re mRNA levels were significantly increased (P , 0.01) by neonatal exposure to estrogen. Finally, down-regulation of testicular Ob-R gene expression by homologous and heterologous signals was demonstrated as relative levels of Ob-R and Ob-Rb mRNAs were significantly decreased (P , 0.01), in a coordinate manner, in rat testis after exposure to human recombinant leptin in vitro, and after stimulation with hCG and FSH in vivo. In conclusion, our results indicate that testicular Ob-R gene expression is developmentally regulated, imprinted by the neonatal endocrine milieu, and sen

Molecular characterization of an endopolygalacturonase from Fusarium oxysporum expressed during early stages of infection.

Abstract: The tomato vascular wilt pathogen Fusarium oxysporum f. sp. lycopersici produces an array of pectinolytic enzymes that may contribute to penetration and colonization of the host plant. Here we report the isolation of pg5, encoding a novel extracellular endopolygalacturonase (endoPG) that is highly conserved among different formae speciales of F. oxysporum. The putative mature pg5 product has a calculated molecular mass of 35 kDa and a pI of 8.3 and is more closely related to endoPGs from other fungal plant pathogens than to PG1, the major endoPG of F. oxysporum. Overexpression of pg5 in a bacterial heterologous system produced a 35-kDa protein with endoPG activity. Accumulation of pg5 transcript is induced by citrus pectin and D-galacturonic acid and repressed by glucose. As shown by reverse transcription-PCR, pg5 is expressed by F. oxysporum in tomato roots during the initial stages of infection. Targeted inactivation of pg5 has no detectable effect on virulence toward tomato plants.

A fruit‐specific and developmentally regulated endopolygalacturonase gene from strawberry (Fragaria x ananassa cv. Chandler)

Abstract: A fruit-specific and developmentally regulated poly- galacturonase gene (spG gene) from strawberry (Fragaria 3ananassa cv. Chandler) has been cloned and characterized at a molecular and physiological level. Comparison analysis of the corresponding deduced sPG protein have shown that this straw- berry gene is similar to Clade A endopolygalac- turonase genes. Moreover, the spatio-temporal and hormonal gene expression pattern suggests a close relationship between the expression of this gene and the onset of the strawberry fruit ripening process and agrees with that of the production of oligosac- charins which have already been described as active molecules involved in fruit ripening. The results are discussed in terms of a putative role of this enzyme in the release of oligosaccharins from the strawberry fruit cell wall.

Detection of the nondefoliating pathotype of Verticillium dahliae in infected olive plants by nested PCR

Abstract: Spread of Verticillium wilt into newly established olive orchards in Andalucia, southern Spain, has caused concern in the olive industry in the region. This spread may result from use of Verticillium dahliae-infected planting material, which can extend distribution of the highly virulent, defoliating (D) pathotype of V. dahliae to new areas. In this study, a molecular diagnostic method for the early in planta detection of D V. dahliae was developed, aimed especially at nursery-produced olive plants. For this purpose, new primers for nested PCR were designed by sequencing a 992-bp RAPD marker of the D pathotype. The use of the specific primers and different nested-PCR protocols allowed the detection of V. dahliae pathotype D DNA in infected root and stem tissues of young olive plants. Detection of the pathogen was effective from the very earliest moments following inoculation of olive plants with a V. dahliae pathotype D conidia suspension as well as in inoculated, though symptomless, plants.

Epoxidation of limonene over hydrotalcite-like compounds with hydrogen peroxide in the presence of nitriles

Abstract: This paper reports the synthesis of an Mg/Al double layered hydroxide (LDH) by coprecipitation, and of its dodecylsulphate (DS) and dodecylbenzenesulphonate (DBS) intercalates by rehydration. The resulting LDHs, also known as hydrotalcite-like compounds, were characterized by using various instrumental techniques including X-ray diffraction (XRD), IR spectroscopy and solid-state NMR spectroscopy. The solids obtained were tested as catalysts in the epoxidation of limonene with hydrogen peroxide. Because the presence of a nitrile was found to be indispensable for the reaction to develop to an acceptable extent, the influence of various nitriles on catalytic activity was examined. The epoxidation reaction yielded limonene oxide.

Nuclear structure functions and heavy flavour leptoproduction off the nucleus at small x in perturbative QCD

Abstract: Although their coefficient functions often make the studies very hard, the variable-coefficient nonlinear evolution equations (vcNLEEs) are of current interests since they are able to model the real world in many fields of physical and engineering sciences. In this paper, based on the computerized symbolic computation, a variable-coefficient balancing-act method is proposed. Being concise and straightforward, it can be applicable to certain vcNLEEs, to get the solitonic features out, along with other exact analytic solutions, all beyond the travelling waves. By virtue of the method, such new solutions are demonstrated for a variable-coefficient KdV equation arising from fluid dynamics, plasmas and other fields. Special attention is paid to the one- and two-soliton-type solutions. Sample applications and physical interests are discussed, such as coastal waters of the world oceans, plasma physics, liquid drops and bubbles. Nonlinear interaction hallmarked by the phase shifts is pictured. Comparisons are made with other results in the literature.