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At least 19 recordsLinked to original sources

Effect of various essential oils isolated from Douglas fir needles upon sheep and deer rumen microbial activity.

The effects of essential oils isolated from Douglas fir needles on sheep and deer rumen microbial activity were tested by use of an anaerobic manometric technique. Rumen microorganisms were obtained from a sheep which had been fed mainly on alfalfa hay and dried range grass. One deer used in this study had access to Douglas fir trees the year around, whereas the other deer had no access to Douglas fir. All of the monoterpene hydrocarbons isolated from Douglas fir needles-alpha-pinene, beta-pinene, limonene, myrcene, camphene, Delta(3)-carene, and terpinolene-promoted only slightly or had no effect on deer rumen microbial activity, whereas all of them promoted activity in sheep rumen microbes, except Delta(3)-carene and terpinolene, which inhibited activity. Of the oxygenated monoterpenes, all monoterpene alcohols-alpha-terpineol, terpinen-4-ol, linalool, citronellol, and fenchyl alcohol-strongly inhibited the rumen microbial activity of both sheep and deer. Monoterpene esters (bornyl acetate) produced mild inhibition for both sheep and deer microbes, and citronellyl acetate inhibited rumen microbial activity in sheep, whereas it promoted activity in both deer. Monoterpene aldehyde (citronellal) inhibited the activity of rumen microbes from both sheep and deer having no access to Douglas fir from the Hopland Field Station, whereas they produced no effect upon the deer having access to Douglas fir from the Masonite forest. Rumen microbial activity for sheep and deer was promoted slightly with aliphatic ester (ethyl-n-caproate). There was a marked difference between sheep and deer rumen microbes as affected by addition of the various essential oils. The monoterpene hydrocarbons promoted activity more on sheep rumen microbes than on deer, and the monoterpene alcohols inhibited sheep rumen microbial activity more than that of deer. Furthermore, the deer rumen microbes from Hopland Field Station were affected more than the deer from Masonite forest.

Animals

Time-course transcriptome and proteomic dynamics during the de novo shoot organogenesis in Chinese fir (Cunninghamia lanceolata).

De novo shoot organogenesis (DNSO) enables plants to regenerate shoots from various explants, offering valuable opportunities for research and plant biotechnology applications. While significant progress has been made in understanding regeneration in angiosperms, the regulatory mechanisms in gymnosperms, particularly Chinese fir (Cunninghamia lanceolata), remain poorly understood, despite its importance as a key timber species in China. This study successfully established an efficient DNSO protocol for Chinese fir, identifying six distinct stages in the process through cellular-level analysis. Time-course transcriptome and proteomics analyses revealed dynamic changes in mRNA and protein levels during regeneration. Notably, proteins showed more significant alterations across a broad range of biological processes, often independent of corresponding mRNA changes. Key pathways associated with ethylene metabolism and abiotic stress responses were enriched, highlighting their critical roles in regeneration. Further experiments confirmed that moderate osmotic stress treatments (150 mm mannitol) and ethylene treatment (100 μm ACC and 5 μm AgNO3) substantially enhanced DNSO efficiency. In summary, this study uncovers the molecular mechanisms underlying Chinese fir DNSO, providing valuable insights into improving plant regeneration efficiency in this economically important species. These findings contribute to advancements in plant biotechnology and sustainable forestry practices.

Cunninghamia

Dispersed repetitive sequences in the chloroplast genome of Douglas-fir.

Restriction mapping and DNA sequencing were used to characterize dispersed repetitive DNA in the chloroplast genome of Douglas-fir [Pseudotsuga menziesii (Mirb.) Franco]. To map repeat families, chloroplast DNA (cpDNA) clones were hybridized at high stringency to one another and to cpDNA cut with restriction enzymes. Repeats are clustered in four regions of the genome and comprise at least six families. Sequence analysis of one repeat family shared among three XbaI fragments indicated the presence of a 633 bp inverted repeat which contains a complete tRNA-Serine (GCU) gene and a highly conserved open reading frame (ORF 3.6). Both ends of this 633 bp dispersed repeat have a transposon-like combination of short direct and inverted repeats. One copy of the repeat flanks one of the endpoints of a major inversion which differentiates Douglas-fir from tobacco cpDNA. Dispersion of repetitive DNA by transposition, coupled with loss of the large inverted repeat, appears to have predisposed conifer cpDNA to a number of inversions. An 8 bp (CATCTTTT) direct repeat in tobacco is located between two inverted sections in Douglas-fir; it may be a target sequence for homologous recombination.

Base Sequence

Antigenic cross-reactivity among monoterpene cyclases from grand fir and induction of these enzymes upon stem wounding.

A major wound response in grand fir (Abies grandis) sapling stems is the rapid increase in monoterpene production at the site of injury. Monoterpene cyclases (synthases) catalyze the formation of monoterpenes from geranyl pyrophosphate, and total cyclase activity increases markedly on wounding. At least six distinct cyclases, producing different monoterpene products, have been isolated from wounded grand fir saplings and characterized. The predominant wound-inducible cyclase produces both alpha- and beta-pinene. This pinene cyclase was purified, and polyclonal antibodies were generated in rabbits against the sodium dodecyl sulfate-denatured protein. The antibody preparation was found to cross-react by Western blotting with other grand fir monoterpene cyclases that produce different olefinic products, but not with monoterpene cyclases from related conifer species (Pinus contorta and P. ponderosa) or from angiosperms (Mentha piperita and M. spicata). The increase in monoterpene cyclase activity after wounding was closely correlated with the appearance of new cyclase protein as determined by immunoblotting. These results indicate that the wound-dependent increase in monoterpene cyclase activity is a consequence of de novo synthesis of cyclase protein.

Antibodies

Two-dimensional gel electrophoresis of proteins for genetic studies in Douglas fir (Pseudotsuga menziesii).

A method of two-dimensional gel electrophoresis of proteins from Douglas fir needles is described. Extraction in the presence of sodium dodecyl sulfate (SDS) and 2-mercaptoethanol followed by heating at 100 degrees C produces reliable two-dimensional gels which are convenient for genetic studies. Three genotypes from different geographical origins have been compared: among 225 loci expressed, 22 display regulatory variations and 7 show allelic variations. Thus it is now possible to undertake the genetic study of Douglas fir using this powerful technique.

Genetic Variation

Force interval relationship (FIR) related to the global function of the left ventricle: a computer study.

A model which relates the left ventricular (LV) geometry, structure and sarcomere properties to its global function, recently proposed by the authors, is extended to account for contractility changes which are a function of the heart rate, prematurity of the beat and calcium transients within the cell. To characterise LV function and relate it to fibre function under varying rhythm conditions, a model of muscle force restitution, based on calcium kinetics, was used to calculate the maximum fibre stress at the optimum sarcomere length sigma o as the parameter which depends on the heart rate, the test pulse interval TPI, the action potential duration APD and the restitution time constant. The global LV force interval relationship FIR was then calculated, and by comparing the calculated FIR to the experimental measurement (in dogs) at the ventricular level, the constants of the restitution of force at the fibre level were derived. Based on these constants, the LV function under ejecting conditions at various rhythm disturbances was calculated and related to the local, distributed parameters. This approach provides a tool to describe ventricular function as well as transmural distribution of stress and sarcomere length at a wide variety of loading and rhythm conditions based on given 'muscle level' parameters.

Biomechanical Phenomena

Genetic analysis of forest tree populations: isolation of DNA from spruce and fir apices.

A method is described for the isolation of DNA from spruce and fir, starting with 3 to 5 apices (5 mg material). Apices are prepared manually from dormant buds harvested in summer and autumn, which are homogenized in 30 microliters buffer containing 1% SDS. The DNA is extracted with phenol and precipitated with ethanol. Agarose gel electrophoresis and Southern hybridization show that its molecular length is ca. 30-40 kb and that it is readily digested with various restriction enzymes. The method is very fast, it does not need CsCl centrifugation and is therefore suited for the analysis of large numbers of individual trees. Moreover, the buds can be collected all over the year. The yield of the method is up to 30 micrograms of high molecular weight DNA, enough to do several digests and hybridizations.

DNA

Organogenesis in in vitro cultures of embryonic shoots of Abies balsamea (balsam fir).

Embryonic shoots of 15- to 20-year-old Abies balsamea (balsam fir) trees were soaked in (a) water for 15 min or 24 hr and (b) water with 1000 mg per 1 indolebutyric acid (IBA), N-dimethylaminosuccinamic acid (Alar-85), or 1-phenyl-3-methyl-5-pyrazolone (PPZ), singly or in combination with 100 mg per 1 caffeic acid, for 15 min. After the soaking, the embryonic shoots were transferred to a nutrient medium. Nonsoaked (control) embryonic shoots elongated and often formed a basal callus but never showed organogenesis. The soaked embryonic shoots formed new apical buds, with or without bud scales, adventitious dwarf needles or shoots, and root- and embryo-like structures. One of the embryos germinated and formed an irregular shoot. No differences were found between the various soak treatments, except that the 15-min water soak was ineffective. The 24-hr water soak was as effective as the 15-min growth regulator treatments.

Butyrates

Characterization of the constitutive and wound-inducible monoterpene cyclases of grand fir (Abies grandis).

Monoterpene cyclase activity is greatly increased in grand fir (Abies grandis) sapling stems in response to wounding and the composition of the cyclic olefin mixture generated differs from that produced constitutively as determined by radio gas-liquid chromatography. Cell-free extracts from wounded stems and from non-wounded controls were systematically compared for monoterpene cyclase activities following partial purification and separation of these enzymes by anion-exchange chromatography (Mono Q FPLC) and native PAGE. The increase in monoterpene cyclase activity following wounding represents both the apparent enhancement of constitutive cyclase activities and the appearance of novel cyclization enzymes that are absent in nonwounded controls. A pinene cyclase was shown to be the major wound-inducible enzyme directly responsible for oleoresin monoterpene formation and was tentatively identified as a 62-kDa protein by SDS-PAGE.

Enzyme Induction

Wound-inducible pinene cyclase from grand fir: purification, characterization, and renaturation after SDS-PAGE.

The major wound-inducible monoterpene synthase (cyclase) of grand fir (Abies grandis) stems transforms geranyl pyrophosphate to both (-)-alpha-pinene (40%) and (-)-beta-pinene (60%). The enzyme was purified to apparent homogeneity by anion-exchange and hydrophobic interaction chromatography, coupled to discontinuous native polyacrylamide gel electrophoresis at neutral pH and polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate (also at neutral pH) followed by renaturation in 1% Tween 20 (polyoxyethylenesorbitan monolaurate). The renatured enzyme produced a mixture of isomeric pinenes from geranyl pyrophosphate identical to that generated by the native form. The protein exhibited a molecular weight of 63,000 by gel permeation chromatography and of 62,000 by denaturing gel electrophoresis, indicating that the monomer is active. The enzyme required Mn2+ (Km = 30 microM) for activity, exhibited a Km value of 6 microM for the substrate geranyl pyrophosphate, showed a pH optimum at 7.8 and temperature optimum at 42 degrees C, and was inhibited by pyrophosphate (I50 = 0.17 mM), orthophosphate (I50 = 51 mM), and alpha-pinene, as well as by the histidine-directed reagent diethylpyrocarbonate (I50 = 0.64 mM) and the cysteine-directed reagent p-hydroxymercuribenzoate (I50 = 1.9 microM). Although similar in many respects to constitutive monoterpene cyclases of herbaceous species, this inducible cyclase, the first enzyme of this type to be purified to homogeneity from a conifer, is distinguished by the relatively high pH optimum, and the strict specificity and high affinity for the divalent metal ion cofactor.

Cations, Divalent

Changes in enzymic activities of nucleoside diphosphate sugar interconversions during differentiation of cambium to xylem in pine and fir.

A protein fraction [precipitate obtained between 40 and 65% (NH4)2SO4 satn.] prepared from cambial cells, differentiating xylem cells and differentiated xylem cells of pine and fir trees contained all the enzymes required for the nucleoside diphosphate sugar interconversions. By using UDP-D-[U-14C]glucose or UDP-D-[U-14C]galactose, UDP-D-[U-14C-]glucuronic acid and UDP-D-[U-14C]xylose as substrates, the activities of UDP-D-galactose 4-epimerase (DC 5.1.3.2), UDP-D-xylose 4-epimerase(EC 5.1.3.5), UDP-D-glucose dehydrogenase (EC 1.1.1.22) and UDP-D-glucuronate 4-epimerase (EC5.1.3.6), UDP-d-glucuronate decarboxylase (EC 4.1.1.35) were measured at different stages of cell-wall development. The specific activities and the activities per cell of these enzymes varied during differentiation of cambium to xylem according to the type polysaccharide synthesized. Variations were also found between the two species investigated. These data, compared with those obtained in out previous work on angiosperms [see the preceding paper, Dalessandro & Northcote (1977) Biochem. J. 162, 267-279], suggest that some control of polysaccharide synthesis operates at the level of the formation of the precursors of pectin and hemicellulose syntheses.

Carbohydrate Epimerases

[Impact of the use of larvivorous fish Poecilia reticulata on the transmission of malaria in FIR of Comoros].

Field tests were conducted in the Grande Comore Island, Federal Islamic Republic of Comoros, in order to evaluate the potential of the larvivorous fish Poecilia reticulata for the control of the malaria vector Anopheles gambiae s.s. Due to the high permeability of soil, Anopheles breeding sites in all island occurs only in the man-made water reservoirs. The study was carried out from November 1987 to November 1988 within a framework of a malaria and filariasis control programme, supported by WHO and UNDP in collaboration with the Government of the FIR of Comoros. All larval breeding places of An. gambiae existing in the village of Hantsambou were recorded (59 ablutions basins and 61 cisterns) and provided initially with 3-5 specimens of P. reticulata/m3 in November 1987, after the importation of the larvivorous species from Mayotte Island. The percentage of breeding places positive for An. gambiae decreased from 41% to 6% after one year. Pyrethrum spray catch showed a reduction of indoor resting density from 5.5 to 0.3, while the ma value, number of Anopheles bites/man/night, obtained by night-biting catches, decreased from 6.3 to 1.2. At the same time of the reduction of entomological indices parasite index for P. falciparum and spleen rate drop steadily in 5-9 years school children. The tested vector control method, well accepted by the community, could be implemented in malaria control through primary health care, being the ecological conditions in the entire island very peculiar.

Animals

[Antibody levels against circumsporozoitic protein of Plasmodium falciparum and their use as epidemiological indicators of malaria transmission in FIR of Comoros].

Epidemiological survey on prevalence of Plasmodium falciparum anti-circumsporozoite antibodies (Ab-CS) was carried out in 21 villages of Comoros FIR, at the beginning of 1988 rainy season. Evaluation of anopheline indoor resting densities was also carried out at the same time. Frequencies of antibody-positive subjects vary considerably in the different villages, according to the Anopheles gambiae and A. funestus densities per room, which are determined by ecological factors. Ab-CS prevalence varies in the sample population from 5.5% in 3-4 years children to 40% in those of 5 years. Starting from 6-7 years group, prevalence increased steadily reaching a plateau by 30 years of age. The detection of Ab-CS levels in a sample population is a good tool to evaluate malaria transmission levels, especially where the epidemiological situation does not allow a reliable entomological evaluation.

Adolescent

Cardiopulmonary responses after spontaneous inhalation of Douglas fir smoke in goats.

In the majority of clinical cases, smoke inhalation results in a self-limited lung injury mostly confined to the airways. In this study, an animal model of inhalation injury was developed that reflected similar pathophysiology. Cardiopulmonary parameters were studied in awake, instrumented goats following spontaneous inhalation of characterized Douglas fir smoke. Peak carboxyhemoglobin levels averaged 37% during a mean exposure time of 33 minutes. All animals survived the 24-hour study period, and showed only transient abnormalities in lung fluid balance and gas exchange, with no change in lung mechanics or plasma eicosanoid (TxB2 and 6-keto-PGF1 alpha) levels. However, extravascular lung water at 24 hours was increased 33%, suggesting the presence of some airway edema and retained secretions. We feel this model fairly represents the majority of clinical smoke inhalation cases. This model is compared to other large animal inhalation injury models producing more severe lung injury.

6-Ketoprostaglandin F1 alpha

Enrichment of cadmium-mediated antibiotic-resistant bacteria in a Douglas-fir (Pseudotsuga menziesii) litter microcosm.

A set of Douglas-fir needle litter microcosms was amended with cadmium, acid, a combination of both, or neither. After 2 weeks of incubation, bacterial colony counts were made of litter homogenates inoculated onto agar media containing an antibiotic (streptomycin, chloromycetin, ampicillin, or gentamicin), cadmium, both, or neither. In all microcosms bacterial abundance was similar but the quality was very dissimiliar. Cadmium-treated microcosms had populations enriched for cadmium and gentamicin resistance and streptomycin and chloramphenicol sensitivity. Acid amendment had no consistent effect on the microcosm populations except that which could be attributed to the cadmium treatment amendment alone.

Anti-Bacterial Agents