Showing papers by "Institut national de la recherche agronomique published in 1989"

The Effects of Dietary α-Linolenic Acid on the Composition of Nerve Membranes, Enzymatic Activity, Amplitude of Electrophysiological Parameters, Resistance to Poisons and Performance of Learning Tasks in Rats

Abstract: Feeding rats diets containing oils that have a low alpha-linolenic acid [18:3(n-3)] content, such as sunflower oil, results in reduced amounts of docosahexaenoic acid [22:6(n-3)] in all brain cells and organelles compared to rats fed a diet containing soybean oil or rapeseed oil. During the period of cerebral development there is a linear relationship between the n-3 fatty acid content of the brain and that of food until alpha-linolenic acid represents approximately 200 mg/100 g food [0.4% of the total dietary energy for 18:3(n-3)]. Beyond that point brain levels reach a plateau. Similar values are also found for other organs. The level of 22:6(n-3) in membranes is little affected by the dietary quantity of linoleic acid [18:2(n-6)] if 18:3(n-3) represents approximately 0.4% of energy. In membranes from rats fed diets containing sunflower oil, Na+, K(+)-ATPase activity in nerve terminals was 60%, 5'-nucleotidase in whole brain homogenate was 80%, and 2',3'-cyclic nucleotide 3'-phosphodiesterase was 88% of that in membranes from rats fed diets containing soybean oil. A diet low in alpha-linolenic acid leads to anomalies in the electroretinogram, which partially disappear with age. It has little effect on motor activity, but it seriously affects learning tasks as measured with the shuttle box test. Rats fed a diet low in alpha-linolenic acid showed an earlier mortality in response to an intraperitoneal injection of a neurotoxin, triethyltin, than did rats fed a normal soybean oil diet.

Comparison of butyrylcholinesterase and acetylcholinesterase

Abstract: Acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) are compared, the first focus being on the high homologies of the molecular forms, and the homologies in protein sequences. Cholinesterases are the prototype of a new family of related serine hydrolases. Secondly, the distribution and regulation of AChE and BChE is reviewed. In this context the proposed noncholinergic roles for cholinesterases are described. Finally, comparison of the structure and base composition of the genes may give clues to understanding the origin and evolution of AChE and BChE

Structure and activity of proteins from pathogenic fungi Phytophthora eliciting necrosis and acquired resistance in tobacco.

Abstract: The phytopathogenic fungi Phytophthora cryptogea and Phytophthora capsici cause systemic leaf necrosis on their non-host tobacco; in culture they release proteins, called cryptogein and capsicein, which elicit similar necrosis. In addition, both proteins protect tobacco against invasion by the pathogen Phytophthora nicotianac, the agent of the tobacco black shank, that is unable to produce such an elicitor. Cryptogein causes visible leaf necrosis starting at about 1 microgram/plant, whereas 50-fold as much capsicein is required for the same reaction. Capsicein induces protection even in near absence of leaf necrosis. The activities of both elicitors are eliminated upon pronase digestion. They are proteins of similar Mr (respectively 10,323 and 10,155) and their complete amino acid sequences were determined. They consist of 98 residues, with some internal repetitions of hexapeptides and heptapeptides. 85% identity was observed between both sequences: only two short terminal regions are heterologous, while the central core is entirely conserved. Secondary structure predictions, hydropathy and flexibility profiles differ only around position 15 and at the C-terminus; these modifications could play a role in the modulation of their biological activities. After a search of the sequence data bases, they appear to be novel proteins.

Protoporphyrinogen oxidase as a molecular target for diphenyl ether herbicides.

Abstract: Diphenyl ether herbicides induce an accumulation of protoporphyrin IX in plant tissues. By analogy to human porphyria, the accumulation could be attributed to decreased (Mg or Fe)-chelatase or protoporphyrinogen oxidase activities. Possible effects of acifluorfen-methyl on these enzymes were investigated in isolated corn (maize, Zea mays) etioplasts, potato (Solanum tuberosum) and mouse mitochondria, and yeast mitochondrial membranes. Acifluorfen-methyl was strongly inhibitory to protoporphyrinogen oxidase activities whatever their origins [concn. causing 50% inhibition (IC50) = 4 nM for the corn etioplast enzyme]. By contrast, it was roughly 100,000 times less active on (Mg or Fe)-chelatase activities (IC50 = 80-100 microM). Our results lead us to propose protoporphyrinogen oxidase as a cellular target for diphenyl ether herbicides.

Nucleotide sequence of potato virus Y (N Strain) genomic RNA.

Abstract: SUMMARY The complete nucleotide sequence of the genomic RNA of the potyvirus potato virus Y strain N (PVYn) was obtained from cloned cDNAs. This sequence is 9704 nucleotides long and can encode a polyprotein of 3063 amino acids. The positions of the cleavage sites at the N terminus of the capsid and cytoplasmic inclusion proteins have been determined. Other putative protein cleavage sites have been deduced by searching for consensus sequences and by analogy with the polyprotein of the tobacco vein mottling virus and of the tobacco etch virus. Comparison of the PVY polyprotein sequence with that of other potyvirus polyproteins shows similarities in genome organization and a high level of identity along most of the polyprotein, except for the putative proteins flanking the helper component. A search for specific protein motifs has revealed the existence of a potential metal-binding site at the putative N terminus of the helper component in potyviruses. The possible functions of this structure are discussed.

Functional evidence for an auxin receptor at the plasmalemma of tobacco mesophyll protoplasts.

Abstract: Tobacco mesophyll protoplasts were previously shown to respond to naphthaleneacetic acid by modifying their transmembrane potential difference. In the present work, evacuolated protoplasts were used to show that this response resides only at the plasmalemma. This electrical response was investigated by using polyclonal antibodies directed against plasma membrane antigens presumably involved in the reception and transduction of the auxin signal. An IgG fraction from an antiserum directed against the membrane auxin-binding protein from maize coleoptile completely inhibited the naphthaleneacetic acid-induced response of tobacco protoplasts. The suppression of the auxin-induced variation in the transmembrane potential difference by an IgG preparation directed against the plasmalemma ATPase from yeast demonstrated the involvement of the ATPase in the electrical response. Variation induced by fusicoccin in the transmembrane potential difference of tobacco protoplasts was unaffected by the anti-auxin-binding protein IgG fraction but was completely suppressed by the anti-ATPase IgG preparation. These results demonstrate the presence of a membrane receptor for auxin at the plasmalemma, the binding of the hormone to this receptor leading to the activation of the proton-pumping ATPase. They also show that at least the primary steps of activation by naphthaleneacetic acid are distinct from those of the fusicoccin-induced response.

Estimation of sow milk nutrient output.

Abstract: Ten replicates of two littermate gilts were used during a 21-d lactation in order to calculate relationships between milk nutrient intake and piglet growth rate and composition of gain. Gilts were fed 14.2 or 10.4 Mcal ME/d and litter size was standardized to 9 or 10 piglets. Piglets had no access to creep feed. Milk production was measured on 10 sucklings over 12 h on d 1, 5, 9, 13, 17 and 21 by the weigh-suckle-weigh method. Heat production of the piglets was measured (RQ method) on the same days in a confinement chamber. Milk composition was determined on the days following milk production measurements. Four to ten piglets/litter were slaughtered at weaning and their body composition was determined. Milk nutrient production during part of lactation was related closely to piglet weight gain and body weight (R2 = .80 to .96). Milk DM, energy and N output over the entire lactation were predicted from piglet ADG (R2 = .87 to .90) when, for each litter, the difference between energy in piglet daily weight gain measured by the slaughter technique and energy in piglet daily weight gain estimated by the RQ method was included in the model. This variable corrects for milk production measurement errors. The relationships were slightly improved, especially for energy output, when the composition of piglet weight gain was taken into account (R2 = .93 to .97).

A simulation model of the three-dimensional architecture of the maize root system

Abstract: In order to study the nutrient and water uptake of roots in situ, we need a quantitative three-dimensional dynamic model of the root system architecture. The present model takes into account current observations on the morphogenesis of the maize root system. It describes the root system as a set of root axes, characterised by their orders and their inter-node of origin. The evolution of the simulated pattern is achieved by three processes, occuring at each time step: emission of new primary root axes from the shoot, growth and branching of existing root axes. The elongation of an axis depends on its order, inter-node and local growing conditions. Branches appear acropetally at a specified distance from the apex and from former branches, along ranks facing xylem poles, with a branching angle specific to their order and inter-node. From the three-dimensional branched patterns simulated by the model, various outputs, such as root profiles or cross-section maps can be computed, compared to observed data and used as inputs in uptake models. A number of examples of such possible outputs are presented.

Biphasic response in the secretion of gonadotrophin-releasing hormone in ovariectomized ewes injected with oestradiol.

Abstract: In ovariectomized ewes, an injection of oestrogen initially inhibits the tonic secretion of LH, and then induces a large release of LH similar to the preovulatory surge in intact ewes. The pattern of hypothalamic secretion of gonadotrophin-releasing hormone (GnRH) into the pituitary portal blood during this biphasic response to oestrogen was investigated in conscious, unrestrained, ovariectomized adult Ile-de-France ewes during the breeding season. The ewes were ovariectomized and implanted with cannulae for portal blood collection on the same day. Seven days later, portal and peripheral blood samples were collected simultaneously every 5 min for 25 h. The ewes were injected with oestradiol-17 beta (25 micrograms i.v. and 25 micrograms i.m.) 6.25 h after the start of sampling. GnRH and LH were measured by radioimmunoassay in portal and jugular plasma samples respectively. A clear pulsatile pattern of LH secretion was observed before the oestradiol injection in all the ewes, followed by the typical biphasic decrease (negative feedback) and increase (positive feedback) in mean concentrations. The sampling period was divided, for analysis, into pretreatment, negative feedback and positive feedback phases. Before injection with oestradiol, the GnRH pulses were clearly defined in portal blood and were always synchronized with LH pulses in the peripheral circulation. The frequency was 5.9 +/- 0.6 pulses/6 h (mean +/- S.E.M.), and the amplitude was 31.6 +/- 7.6 pmol/l. During negative feedback, both the frequency (4.2 +/- 0.5 pulses/6 h, P less than 0.01) and amplitude (15.2 +/- 4.6 pmol/l, P less than 0.05) of the GnRH pulses decreased.(ABSTRACT TRUNCATED AT 250 WORDS)

Identification and sequence analysis of a second form of prolactin receptor by molecular cloning of complementary DNA from rabbit mammary gland.

Abstract: Two lambda gt11 clones containing fragments of cDNA encoding the prolactin receptor from rabbit mammary gland were isolated using a rat liver prolactin receptor cDNA probe. An 1848-base-pair open reading frame encodes a mature prolactin-binding protein of 592 amino acids that contains three domains: (i) the extracellular, amino-terminal, prolactin-binding region of 210 residues; (ii) the transmembrane region of 24 residues; and (iii) the intracellular, carboxyl-terminal domain of 358 residues. This latter domain is much longer than the cytoplasmic domain (57 residues) previously described for the rat liver prolactin receptor. In addition, the sequence identity of this form of prolactin receptor with the growth hormone receptor is extended in the cytoplasmic domain.

Influence of a fungal polysaccharide, scleroglucan, on clay microstructures

Abstract: The microorganization of complexes formed between clay minerals (Ca-kaolinite, Ca-montmorillonite) and a fungal polysaccharide (scleroglucan) was studied with water content and apparent volume measurements. X-ray diffractometry and scanning electron microscopy. Scleroglucan did not appear to intercalate montmorillonite. The fabric of the complexes was a card-house structure for koalinite and a three-dimensional network of quasicrystals for montmorillonite, both with enhanced porosities. The present results indicate that the strong water-stabilizing effect of fungal polysaccharides occurs without major microstructural rearrangements but could be related to the formation of stable organo-mineral networks.

Tungstate, a Molybdate Analog Inactivating Nitrate Reductase, Deregulates the Expression of the Nitrate Reductase Structural Gene

Abstract: Nitrate reductase (NR, EC 1.6.6.1) from higher plants is a homodimeric enzyme carrying a molybdenum cofactor at the catalytic site. Tungsten can be substituted for molybdenum in the cofactor structure, resulting in an inactive enzyme. When nitratefed Nicotiana tabacum plants were grown on a nutrient solution in which tungstate was substituted for molybdate, NR activity in the leaves decreased to a very low level within 24 hours while NR protein accumulated progressively to a level severalfold higher than the control after 6 days. NR mRNA level in molybdate-grown plants exhibited a considerable day-night fluctuation. However, when plants were treated with tungstate, NR mRNA level remained very high. NR activity and protein increased over a 24-hour period when nitrate was added back to N-starved molybdate-grown plants. NR mRNA level increased markedly during the first 2 hours and then decreased. In the presence of tungstate, however, the induction of NR activity by nitrate was totally abolished while high levels of NR protein and mRNA were both induced, and the high level of NR mRNA was maintained over a 10-hour period. These results suggest that the substitution of tungsten for molybdenum in NR complex leads to an overexpression of the NR structural gene. Possible mechanisms involved in this deregulation are discussed.

Distribution of bacteria in the rumen contents of dairy cows given a diet supplemented with soya-bean oil

Abstract: 1. Liquid-associated bacteria (LAB) were harvested from the liquid phase (LAB1) and from the solid phase of rumen contents after washing and manual shaking (LAB2). Solid-adherent bacteria (SAB) were recovered after washing and pummelling the total particles (SAB1). The distribution and the chemical composition of these three bacterial compartments were investigated in four dairy cows fitted with rumen fistulas. The animals received successively a diet consisting of one part hay and one part barley-based concentrate (diet C) and the same diet containing free soya-bean oil (79 g/kg dry matter (DM); diet So).2. The efficiency of removal of SABI from total particles of rumen digesta collected I h after feeding, was calculated from the diaminopimelic acid content in particles and of the corresponding detached bacteria. It was 24% on diet C and 18% on diet So (P < 0.05), using a combination of homogenizing and ‘stomaching’ treatments in saline (9 g sodium chloride/1) (reference treatment). For diets C and So respectively it was lowered by Tween in saline solution (1 g/l; 22.7 and 17.8 %, not significant), but was increased when using a previous chilling (6 h at 4°) of homogenized particles before stomaching in saline (28.8 and 24.7%, P < 0.05) and in Tween 80 in saline (1 g/l; 26.6 and 20.8%, P < 0.05).3. The extent of removal of SABI from the solid fraction of rumen digesta by the reference treatment decreased with decreasing particle size; it was at the highest for particles retained on 4 and 2 mm sieves (62.1–82.1 %) and still elevated for particles retained on 0.8, 0.4 and 0.1 mm sieves (41.3–57.9%). It was very much reduced for particles smaller than 0.1 mm (11.7–14.5 %), suggesting the occurrence of favourable conditions for the adhesion of SAB firmly resistant to removal (SAB2).4. The concentration of total SAB (SABI +SAB2) in particles collected l h after feeding was lower (P < 0.05) in diet C (190 g/kg DM) than in diet So (234 g/kg DM). Values averaged 595–645 g/kg DM for particles smaller than 0.1 mm, but only 61 and 81–98 g/kg DM for particles retained on 4 and 0.4 mm sieves, and on a 0.1 mm sieve respectively. No significant differences were noted between diets but the effect of particle size was highly significant (< 0.1 mm v. others).5. Postprandial variations of concentrations of total SAB on total particles exhibited a large increase I h after feeding in diet So (P < 0.05). Similar but amplified variations were observed for LAB in both diets (P < 0.05).6. Total bacterial mass amounted to 213 and 231 g DM/kg whole-rumen contents DM in diets C and So respectively 6 h after feeding. Mean percentages of total SAB (69.8), LAB1 (7.3) and LAB2 (22.9) in total rumen contents were not significantly modified by the lipid level of the diet.

Rhizobium meliloti fixGHI sequence predicts involvement of a specific cation pump in symbiotic nitrogen fixation.

Abstract: We present genetic and structural analyses of a fix operon conserved among rhizobia, fixGHI from Rhizobium meliloti. The nucleotide sequence of the operon suggests it may contain a fourth gene, fixS. Adjacent open reading frames of this operon showed an overlap between TGA stop codons and ATG start codons in the form of an ATGA motif suggestive of translational coupling. All four predicted gene products contained probable transmembrane sequences. FixG contained two cysteine clusters typical of iron-sulfur centers and is predicted to be involved in a redox process. FixI was found to be homologous with P-type ATPases, particularly with K+ pumps from Escherichia coli and Streptococcus faecalis but also with eucaryotic Ca2+, Na+/K+, H+/K+, and H+ pumps, which implies that FixI is a pump of a specific cation involved in symbiotic nitrogen fixation. Since prototrophic growth of fixI mutants appeared to be unimpaired, the predicted FixI cation pump probably has a specifically symbiotic function. We suggest that the four proteins FixG, FixH, FixI, and FixS may participate in a membrane-bound complex coupling the FixI cation pump with a redox process catalyzed by FixG.

Laboratory experimental study of soil crusting: Relation between aggregate breakdown mechanisms and crust stucture

Abstract: Crusts formation on cores of air-dry and prewetted aggregates was studied, using a laboratory rainfall simulator. Samples were taken out during the experiment, in order to study changes in water content and aggregates size distribution inside the crust and under the crust. Pore space geometry of the crusts is described using light optical microscopy and mercury porosimetry. In the case of air-dry aggregates, aggregates become micro-cracked. Macropores at the surface are quickly closed and ponding occurs rapidly. With prewetted aggregates, microcracking does not occur, and only a very slow abrasion of the aggregates is observed. Porosity remains high and no ponding excess occurs. The discussion of these results shows that it is the way of the wetting and the initial water content which determine the breakdown mechanism, and therefore, the behaviour of the aggregates submitted to rainfall.

Phenotypic and genomic studies of "Cytophaga psychrophila" isolated from diseased rainbow trout (Oncorhynchus mykiss) in France

Abstract: Five strains of gliding bacteria were isolated in France from farmed diseased rainbow trouts reared at low water temperature. The resemblance of these bacteria to the known fish pathogen "Cytophaga psychrophila" led to their comparative study with reference strain NCMB 1947 and with an American isolate. Morphological, physiological, and biochemical characteristics of the seven strains proved to be similar. Comparison of their DNA by the S1 nuclease DNA-DNA hybridization method showed that the seven strains formed a tight genomic species with DNA relatedness above 90%. This is the first identification of this fish pathogen in a European country. The main phenotypic characteristics differentiating this bacterium from other nonpathogenic gliding bacteria of fish origin include a poor gliding movement, yellow compact or weakly rhizoid colonies on solid media, and the presence of flexirubin-type pigments. The inability to metabolize any carbohydrates, the strong proteolytic activity, the absence of growth in more than 0.5% NaCl, and the tolerance to a maximum temperature of 25 degrees C are also useful characteristics of this group of bacteria.

Effect of Sugar Transport Inactivation in Saccharomyces cerevisiae on Sluggish and Stuck Enological Fermentations.

Abstract: Sluggish and stuck (i.e., very delayed or incomplete) fermentations have been often observed in wine making. Some of them appeared to be associated with insufficient levels of yeast nutrients such as assimilable nitrogen. In these conditions, sugar transport catabolite inactivation, which is triggered by the protein synthesis arrest, may account in part for the inhibition of fermentation. Moreover, this mechanism of inhibition may explain the failure of added ammoniacal nitrogen to nitrogen-limited musts to restore or elevate rate of fermentation after the early yeast growth phase.

Factors affecting the spectral response of forest canopies: A review

Abstract: Interpreting remote sensing data on forest canopies demands an adequate knowledge of factors affecting their optical properties. After a short analysis of the optical properties of a forest canopy, a review of the factors acting on the forest reflectance is presented. These factors can be external or internal. The five external factors considered are: size of the viewed area, orientation and inclination of the view axis, sun elevation, nebulosity and wind speed. Three internal factors can also affect forest reflectance: row orientation (for young artificial forests), optical properties of the background (soil and understory), and canopy geometry. The effects of these different factors are analysed and discussed.

Catecholamine-containing neurons in the sheep brainstem and diencephalon: immunohistochemical study with tyrosine hydroxylase (TH) and dopamine-beta-hydroxylase (DBH) antibodies.

Abstract: The present study describes the distribution and morphological characteristics of neurons and nerve fibers containing the catecholamine-synthesizing enzymes, tyrosine hydroxylase and dopamine-beta-hydroxylase, in the sheep brainstem and diencephalon on the basis of immunohistochemical procedures. Neurons and fibers were considered to be dopaminergic if they showed anti-tyrosine hydroxylase immunoreactivity, without corresponding anti-dopamine-beta-hydroxylase immunoreactivity. The structures labeled with both antisera were considered noradrenergic or adrenergic. The distribution of catecholaminergic neurons corresponds to that described by other authors with similar methods in the rat and in primates. The noradrenergic neurons belong to cell groups A1 to A7 and the dopaminergic neurons to cell groups A8 to A15. In almost all studied areas, the catecholaminergic innervation is similar to that observed in the other species. However, the central catecholaminergic systems of the sheep showed some specific characteristics: (1) groups A3 and A4, described in the rat, were not found, (2) group A14 contains fewer neurons than in the rat, (3) group A15 does not contain a dorsal but only a ventral portion, (4) there is a larger dispersion of neurons within each group, especially A6 and A7, than in rodents, and (5) there is a larger noradrenergic innervation of the catecholaminergic groups than in the other species.

Sequential changes in the expression of mitochondrial protein mRNA during the development of brown adipose tissue in bovine and ovine species. Sudden occurrence of uncoupling protein mRNA during embryogenesis and its disappearance after birth.

Abstract: Samples of adipose tissue were obtained from different sites in bovine and ovine foetuses and newborns. RNA was isolated and analysed using bovine cDNA and ovine genomic probe for uncoupling protein (UCP), cDNA for subunits III and IV of cytochrome c oxidase and cDNA for ADP/ATP carrier. UCP mRNA was characterized for the first time in foetal bovine and ovine adipose tissue. It appeared later than mRNA of cytochrome c oxidase subunit III, and increased dramatically at birth (10-fold). ADP/ATP carrier mRNA was expressed at a lower level but also increased 10-fold at birth. It was demonstrated that UCP mRNA reached its highest level at birth in all bovine adipose tissues studied, except subcutaneous tissue. It disappeared quickly afterwards, being no longer detectable two days after birth. Similar variations were observed in newborn lambs. ADP/ATP carrier mRNA showed the same pattern of expression as UCP mRNA; although it was still lightly expressed two days after birth, it disappeared soon afterwards. Only mRNAs for cytochrome c oxidase subunits III and IV remained at the same level during the first postnatal week. On the basis of these data and of observations reported in the literature a sequence of events for the development of brown adipose cells in vivo is proposed. Soon after birth the perirenal adipose tissue of ruminants, which still contains mitochondria of typical brown adipose tissue morphology and high levels of cytochrome c oxidase mRNA, lacks UCP mRNA. Can it still be considered as brown fat? Ruminant species appear to be attractive models to study both the differentiation of brown adipose tissue and its possible conversion to white fat in large animals.