Poplar (Populus nigra L.) plants transformed with aBacillus thuringiensis toxin gene: insecticidal activity and genomic analysis

Insect-resistant poplar (Populus nigra L.) plants have been produced by infecting leaves withAgrobacterium tumefaciens strains carrying a binary vector containing different truncated forms of aBacillus thuringiensis (B.t.) toxin gene under a duplicated CaMV 35S promoter. Putative transgenic plants were propagated by cuttings at two experimental farms (in Beijing and Xinjiang, China). At 2–3 years after transformation, 17 of them were selected on the bases of insect-tolerance and good silvicultural traits, and evaluated for insect resistance, for the presence of theB.t. toxin DNA fragment (Southern blots and PCR) and for the expression of the transgene (western and northern blots). Somaclonal variation, as suggested by the appearance of permanent changes in the shape of the leaves, was also investigated with molecular tools (RFLP (restriction fragment length polymorphism), RAPD (random amplified polymorphic DNA) and microsatellite DNA).Bioassays withApochemia cineraius andLymantria dispar on the leaves of the selected clones showed different and, in some cases, high levels of insecticidal activity. The molecular analysis demonstrated integration and expression of the foreign gene. Somatic changes were correlated to extensive genomic changes and were quantified in dendrograms, in terms of genomic similarity. The analysis of control plants suggested that genomic changes were correlated to thein vitro culture step necessary forA. tumefaciens-mediated gene transfer, rather than to the integration of the foreign genes.Three transgenic clones (12, 153 and 192), selected for insect resistance, reduced morphological changes and promising silvicultural traits, are now under large-scale field evaluation in six different provinces in China.     

Does photorespiration protect the photosynthetic apparatus in french bean leaves from photoinhibition during drought stress?

Dithiothreitol (DTT), an inhibitor of violaxanthin de-epoxidation and zeaxanthin formation in chloroplasts, inhibited blue-light-stimulated stomatal opening in epidermal peels of Vicia faba L. in a concentration-dependent fashion. Complete inhibition was observed at 3 mM DTT. The DTT effect was specific for the stomatal response to blue light, and the red-light-stimulated opening, which depends on photosynthetic reactions in the guard cells, was unaffected. Preirradiation of stomata in epidermal peels with increasing photon fluence rates of red light, prior to an incubation in 10 mol·m^-2·s^-1 of blue light and 100 mol·m^-2·s^-1 red light, resulted in a DTT-sensitive, blue-light-stimulated opening that was proportional to the fluence rate of the red light pre-treatment. Guard cells in epidermal peels and guard-cell protoplasts irradiated with red light showed increases in their zeaxanthin content that depended on the fluence rate of red light, or on the incubation time. The increases in zeaxanthin concentration were inhibited by DTT. The obtained results indicate that zeaxanthin could function as a photoreceptor mediating the stomatal responses to blue light.Abbreviation DTT dithiothreitol This work was supported by grants from the National Science Foundation and the US Department of Energy to E.Z.     

Laminin and Neuropeptide Y Are Increased by Synapsin Transfection in Cultured NG108-15 Neuroblastoma/Glioma Hybrid Cells

Abstract: We have investigated the presence and expression of laminin and neuropeptide Y (NPY) in several NG108-15 cell lines transfected with synapsin Ib, IIa, or IIb. The content of laminin, a basal membrane glycoprotein that promotes adhesion and induces neurite outgrowth and neuronal differentiation, was increased in all transfected cell lines examined. In cells that were chemically differentiated with prostaglandin E₁ plus 3-isobutyl-1-methylxanthine, laminin levels were increased even further. The content of NPY, suggested to be a neurotransmitter/neuromodulator in peripheral sympathetic neurons as well as in central neurons, was also increased in all transfected cell lines examined. Immunohistochemical analysis combined with confocal laser microscopy showed that NPY staining was granular and very often enriched in neuritic varicosities. The distribution and the staining pattern of NPY were consistent with storage of NPY in large dense-cored vesicles. The results indicate that, in differentiated neurons, the synapsins increase the levels of a neuropeptide transmitter stored in large dense-cored vesicles and of an extracellular matrix protein associated with neuronal maturation.     

Swim Stress Increases the Potency of Glycine at the N-Methyl-d-Aspartate Receptor Complex

Abstract: We have previously demonstrated that chronic administration of antidepressants results in a reduction in the potency of glycine to displace 5,7-[³H]dichlorokynurenic acid (5,7-[³H]-DCKA) from the strychnine-insensitive glycine recognition site of the NMDA receptor complex. We now report that exposure of rats to the forced swim test results in a 56% increase in the potency of glycine to displace 5,7-[³H]DCKA from frontal cortical homogenates. These data are consistent with the hypothesis that the forced swim test, a preclinical screen sensitive to acute administration of antidepressant drugs and NMDA receptor antagonists, also results in adaptation of the NMDA receptor complex. Moreover, these data lend further support to the hypothesis that glutamatergic pathways are involved in the neurobiological response to stress and, potentially, in the pathophysiology of depression.     

Intramolecular domain-domain interactions and intermolecular self-association in bovine prothrombin. A potentiometric and laser light-scattering study

The interaction of bovine prothrombin with Ca²⁺ and Mg²⁺ ions was investigated by following H⁺ release as a function of metal ion concentration at pH 6 and pH 7.4 at high and low ionic strength. Prothrombin Ca²⁺ and Mg²⁺ binding is characterized by high- and low-affinity sites. M²⁺ binding at these sites is associated with intramolecular conformational changes and also with intermolecular self-association. The pH dependence of H⁺ release by M²⁺ is bell shaped and consistent with controlling pK_a values of 4.8 and 6.5. At pH 6 and low ionic strength, both Ca²⁺ and Mg²⁺ titrations following H⁺ release clearly show independent low- and high-affinity binding sites. Laser light scattering reveals that at pH 7.4 and low ionic strength, and at pH 6.0 and high ionic strength, the prothrombin molecular weight is between 73 and 98 kD. At pH 7.4 and high ionic strength, prothrombin is monomeric in the absence of metal ions, but appears to dimerize in the presence of M²⁺. At pH 6.0 and low ionic strength prothrombin exists as a dimer in the absence of metal ions and is tetrameric in the presence of Ca²⁺ and remains dimeric in the presence of Mg²⁺. These results and those for metal ion-dependent H⁺ release indicate that H⁺ release occurs concomitantly with association processes involving prothrombin.Abbreviations GLA -carboxyglutamic acid; fragment 1. amino terminal residues 1–156 of bovine prothrombin - MES 2-(N-morpholino) ethanesulfonic acid - MOPS 3-(N-morpholino) propanesulfonic acid - PS/PC phosphatidylserine/phosphatidylcholine vesicles - ionic strength     

Behavioral adaptations of two phytophagous insects feeding on two species of phototoxic Asteraceae

Two phototoxic plants of the Asteraceae family were studied in relation to species of phytophagous insects for which they are hosts:Argyrotaenia velutinana Wlk. feeding onChrysanthemum leucanthemum L. andChlorochlamys chloroleucaria (Guenée) colonizingRudbeckia hirta L. The toxicity of these two plants is related to the presence of acetylenes and thiophenes that induce a light-mediated production of deleterious singlet oxygen and other free radicals (phototoxicity). Results showed that females ofA. velutinana laid their eggs preferentially in the shade and the larvae adopted hiding behaviors, such as bending of ligulate corollas and silk spinning to build opaque shelters. By avoiding direct exposure to the sun, both behaviors may reduce phototoxicity associated with ingested plant materials. Furthermore, larvae ofC. chloroleucaria demonstrated a preference in the field for pollen, which constitutes a nonphototoxic tissue of their host plant. Experimental alterations of these specific behaviors induced important biological consequences for larvae of both insects such as mortality or reduction of larval growth rate. These results reinforce the idea that behavior may constitute an efficient adaptation to avoid phototoxicity.     

In vitro autoradiographic localization of vasoactive intestinal peptide (VIP) binding sites in the rat central nervous system

This paper describes the autoradiographic distribution of VIP binding sites in the rat central nervous system using monoiodinated ¹²³I-labeled VIP. High densities of VIP binding sites are observed in the granular layer of the dorsal dentate gyrus of the hippocampus, the basolateral amygdaloid nucleus, the dorsolateral and median geniculate nuclei of the thalamus as well as in the ventral part of the hypothalamic dorsomedial nucleus.     

Interaction of peanut agglutinin,a lectin specific for nonreducing terminal d-galactosyl residues,with embryonal carcinoma cells

Peanut agglutinin (PNA), a lectin specific for terminal d-galactosyl residues, was found to react with embryonal carcinoma cells, but not with their differentiated derivatives. Receptors for PNA were detectable at the surface of all cells of the quasinullipotent F9 line and on only 50% of the multipotent PCC3/A/1 line. The fraction of the PCC3/A/1 population which expresses the F9 antigen was found to be included in the subpopulation carrying the PNA receptors. PNA⁺ and PNA^? subpopulations of PCC3/A/1 were separated by a PNA-mediated reversible agglutination of PNA⁺ cells with rabbit erythrocytes. These subpopulations were essentially F9⁺ and F9^?, respectively.     

Potential of ultraviolet radiation for control of american cockroach populations

Populations of Periplaneta americana (L.) were exposed for 8–20 week periods in specially designed rooms to 254 nm UV at low intensity (50–115 ergs sec^–1cm^–2), high intensity (160–220 ergs sec^–1cm^–2), or to white light. The rooms contained tables and chairs to simulate occupied space, with food and water placed in positions exposed to UV radiation. General irradiation (where the whole room was exposed to UV) at 115 ergs sec^–1cm^–2 and above was effective in producing high mortality in all stages except 8–10th instar nymphs and adults. Hot-spots irradiation (where UV lamps were placed behind table and chair harborages) produced high mortality only in 1 st-3rd instar nymphs which would result in slower elimination of a population. Crude aggregation pheromone was not successful in holding cockroaches close to radiation sources or substantially increasing mortality under the conditions of the experiments.

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Zusammenfassung Populationen von Periplaneta americana (L.), die hinsichtlich ihrer Alterszusammensetzung (2.–3.; 5–6.; 8.–10. und adultes Stadium) und der Anzahlen in jedem Stadium festgelegt waren, wurden für 8–20 Wochenperioden in speziell dafür entworfenen Räumen einer 254 nm UV-Bestrahlung mit geringer (50–115 erg sec^–1cm^–2) oder hoher (160–220 erg sec^–1cm^–2) Intensität oder weißem Licht (als Kontrolle) ausgesetzt. Die Räume enthielten Tische und Stühle, um bewohnten Raum mit natürlichen Zufluchtsstätten mit Nahrung und Wasser an Stellen, die der UV-Bestrahlung unterlagen, zu simulieren. Ganzraumbestrahlung mit 115 erg sec^–1cm^–2 und darüber erzeugte hohe Mortalität bei 1.–3. und 5.–6.-Larvenstadien, örtliche Bestrahlung (UV-Lampen hinter Tisch- und Stuhl-Zufluchtsstätten) dagegen nur beim 1.–3.-Stadium, was zu einer langsameren Ausrottung einer Population führen würde. Ungereinigtes Aggregationspheromon als Zusatz, um Schaben dicht an die UV-Quellen zu locken und sie hier zu halten, war offenbar unwirksam, da eben die Mortalität nicht signifikant zunahm. Dieses Versagen war in erster Linie auf die Konkurrenz mit der Fülle von natürlichem Pheromon, das von den gewohnten Zufluchtsstätten ausging, zurückzuführen, verbunden mit der dem UV-Licht innewohnenden Abschreckung. Dennoch darf man annehmen, daß UV-Bestrahlung einen bedeutsamen Wert für die Verhinderung eines Populationswachstums (durch Ausschalten junger Larvenstadien) besitzt, besonders dort, wo chemische Bekämpfung aus Gesundheits- und Sicherheitsgründen oder wegen gesetzlichen Einschränkungen nur begrenzt möglich ist.

     

Extremely rapid degradation of [3H] methionine-enkephalin by various rat tissues in vivo and in vitro

Thirty sec after the intrajugular injection of [³H] methionine-enkephalin (met-enkephalin) in the rat, the radioactivity was already distributed in an apparent volume of 53 ml and the metabolic clearance rate calculated from the characteristics of the plasma disappearance curve was 10 ml/min. As shown by partition chromatography plasma extracts obtained 15 sec after injection of [³H] met-enkephalin, only 5% of the total radioactivity migrated as the intact pentapeptide, while no detectable intact pentapeptide remained 2 min after injection, thus indicating a half-life of [³H] met-enkephalin of the order of 2 to 4 sec. Incubation of rat cerebral tissue with [³H] met-enkephalin indicates that the first step in the breakdown of met-enkephalin in both plasma and brain tissue is cleavage of the Tyr-Gly amide bond. These data offer an explanation for the low activity of met-enkephalin after intraventricular or intravenous administration.