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.     

In vitro peptidergic neurons from the adult locust pars intercerebralis: Morphological and immunocytological studies

Primary cell cultures were prepared from a major neurosecretory center of the adult locust brain, the pars intercerebralis, in order to characterize neurosecretory cells growingin vitro. Individual pars intercerebralis could be removed free of surrounding tissue and dissociated by mechanical treatment. Mature neurosecretory neurons of different sizes regenerate new neurites during the initial three daysin vitro in serum-free medium. They show a tendency to sprout one primary neurite from which fine processes develop. By means of electron microscopy, we observed the integrity of the cellular organelles, indicating that cultured neurons are healthy, and we were able to distinguish three types of neurosecretory neurons on the basis of the ultrastructural aspects of the neurosecretory material. These three types have the same ultrastructural characteristics asin situ neuroparsin, ovary maturing parsin and locust insulin related peptide neurons. Immunogold labelling at the electron microscopic level, using the two available specific antibodies, anti-neuroparsin and anti-ovary maturing parsin, confirms the morphological characterization of neuroparsin and ovary maturing parsin cells. These results show for the first time that cultured locust neurosecretory neurons behave like thosein vivo, in terms of their ultrastructure and immunocytochemistry. Moreover, the presence of recently-formed neurosecretory material both in the Golgi zone of the perikaryon and in the neuronal processes indicates that cultured neurons have functional capacity since they are able to synthesizede novo and to transport the neurosecretory material along the neurite. Thus our well-characterized culture system provides a suitable invitro model to investigate the secretory mechanism of locust neurosecretory neurons.     

组建昆虫系统生物学的目的性和可行性探索

通过作在昆虫生物学方面近五十年研究工作的体会．提出了组建昆虫系统生物学的目的性和可行性探索．拟以昆虫分类序列目、科、属、种为单元,将一些已知种类的有关生物学环节．一一列叙．编成《中国农林昆虫系统生物学手册》．再以目、科、属、种为主体,探求彼此间在生物学各环节间的共性和个性,分别加以比较、归纳和分析．作为这—分支学科的蘸本。     

Gap junctional intercellular communication of bovine luteal cells from several stages of the estrous cycle: Effects of prostaglandin F2α, protein kinase C and calcium

Cellular interactions mediated by both contact-dependent and contact-independent mechanisms are probably important to maintain luteal function. The present studies were performed to evaluate the effects of luteotropic and luteolytic hormones, and also intracellular regulators, on contact-dependent gap junctional intercellular communication (GJIC) of bovine luteal cells from several stages of luteal development. Bovine corpora lutea (CL) from the early, mid and late luteal phases of the estrous cycle were dispersed with collagenase and incubated with no treatment, LH, PGF or LH + PGF (Experiment 1), or with no treatment, or agonists or antagonists of protein kinase C (TPA or H-7) or calcium (A23187 or EGTA; Experiment 2). After incubation, media were collected for determination of progesterone concentrations. Then the rate of GJIC was evaluated for small luteal cells in contact with small luteal cells, and large luteal cells in contact with small luteal cells by using the fluorescence recovery after photobleaching technique and laser cytometry. Luteal cells from each stage of the estrous cycle exhibited GJIC, but the rate of GJIC was least (P<0.05) for luteal cells from the late luteal phase. LH increased (P<0.05) GJIC between small luteal cells from the mid and late but not the early luteal phase. PGF increased (P<0.05) GjIC between small luteal cells from the mid luteal phase and diminished (P<0.05) LH-stimulatory effects on GjIC between small luteal cells from the late luteal phase. Throughout the estrous cycle, TPA decreased (P<0.05) the rate of GjIC between large and small, and between small luteal cells, and A23187 decreased (P<0.05) the rate of GJIC between large and small luteal cells. LH and LH + PGF, but not PGF alone increased (P<0.05) progesterone secretion by luteal cells from the mid and late luteal phases. Agonists or antagonists of PKC or calcium did not affect progesterone secretion by luteal cells. These data demonstrate that both luteal cell types communicate with small luteal cells, and the rate of communication depends on the stage of luteal development. LH and PGF affect GjIC between small luteal cells during the fully differentiated (mid-luteal) and regressing (late luteal) stages of the estrous cycle. In contrast, at all stages of luteal development, activation of PKC decreases GjIC between small and between large and small luteal cells, whereas calcium ionophore decreases GjIC only between large and small luteal cells. Luteotropic and luteolytic hormones, and intracellular regulators, may be involved in regulation of cellular interactions within bovine CL which likely is an important mechanism for coordination of luteal function.     

Response of rice varieties to soil salinity and air humidity: A possible involvement of root-borne ABA

In a phytotron experiment four rice varieties (Pokkali, IR 28, IR 50, IR 31785-58-1-2-3-3) grown in individual pots were subjected to low (40/55% day/night) and high (75/90%) air humidity (RH), while soil salinity was gradually increased by injecting 0, 30, 60 or 120 mM NaCl solutions every two days. Bulk root and stem base water potential (SWP), abscisic acid (ABA) content of the xylem sap and stomatal resistance (rs) of the youngest fully expanded leaf were determined two days after each salt application. The SWP decreased and xylem ABA and rs increased throughout the 8 days of treatment. The effects were amplified by low RH. A chain of physiological events was hypothesized in which high soil electric conductivity (EC) reduces SWP, followed by release of root-borne ABA to the xylem and eventually resulting in stomatal closure. To explain varietal differences in stomatal reaction, supposed cause and effect variables were compared by linear regression. This revealed strong differences in physiological reactions to the RH and salt treatments among the test varieties. Under salt stress roots of IR 31785-58-1-2-3-3 produced much ABA under low RH, but no additional effect of low RH on rs could be found. By contrast, Pokkali produced little ABA, but rs was strongly affected by RH. RH did not affect the relationships EC vs. SWP and SWP vs. ABA in Pokkali, IR 28, and IR 50, but the relationship ABA vs. rs was strongly affected by RH. In IR 31785-58-1-2-3-3 RH strongly affected the relationship SWP vs. ABA, but had no effect on ABA vs. rs and EC vs. rs. The results are discussed regarding possible differences in varietal stomatal sensitivity to ABA and their implications for varietal salt tolerance.     

Gibbard's evolutionary theory of rationality and its ethical implications

Gibbard's theory of rationality is evolutionary in terms of its result as well as its underpinning argument. The result is that judgments about what is rational are analyzed as being similar to judgments of morality — in view of what Darwin suggests concerning the latter. According to the Darwinian theory, moral judgments are based on sentiments which evolve to promote the survival and welfare of human societies. On Gibbard's theory, rationality judgments should be similarly regarded as expressing emotional attachments to behavioral norms which originate and function to coordinate social interaction. Consequently, Gibbard's theory of rationality might be used to illuminate Darwin's theory of morality, and vice versa. Additionally, as argued in the present essay, both can be further elaborated, and defended, by developing related themes in philosophical ethics: viz., connected with Hume and 20th-century emotivists. The main problem is that this general Darwinian approach faces widespread opposition nowadays, not only in ethics but in philosophy of science. The purpose of this essay is to analyze Gibbard's theory, critically and constructively, with emphasis on the pertinent commonalities in Darwin, Hume and the emotivists, while also critically addressing their common enemies. The pervasive methodological orientation is to relate this analysis to (philosophy of) science in general, and biological science in particular.     

Hypothesis on adaptive features of the compound eye of bees: Flower-specific specializations

Summary This note surveys the current knowledge of the existence of acute zones in insects' eyes in relation to beeflower interactions.We suggest some broad generalizations correlating the size, use, structure and presentation of flowers with the foraging behaviour, body size and eye design of bees.Special emphasis is given to the differences between small and large bees and the architecture of flowers that they use.     

Cyclic AMP induces rapid increases in gap junction permeability and changes in the cellular distribution of connexin43

The rapid effects of cAMP on gap junction-mediated intercellular communication were examined in several cell types which express different levels of the gap junction protein, connexin43 (Cx43), including immortalized rat hepatocyte and granulosa cells, bovine coronary venular endothelial cells, primary rat myometrial and equine uterine epithelial cells. Functional analysis of changes in junctional communication induced by 8-bromo-cAMP was monitored by a fluorescence recovery after photobleaching assay in subconfluent cultures in the presence or absence of 1.0 mm 1-octanol (an agent which uncouples cells by closing gap junction channels). Communicating cells treated with 1.0 mm 8-bromo-cAMP alone exhibited significant increases in the percent of fluorescence recovery which were detected within 1–3 min depending on cell type, and junctional communication remained significantly elevated for up to 24 hr. Addition of 1.0 mm 8-bromo-cAMP to cultured cells, which were uncoupled with 1.0 mm octanol for 1 min, exhibited partial restoration of gap junctional permeability beginning within 3–5 min. Identical treatments were performed on cultures that were subsequently processed for indirect immunofluorescence to monitor Cx43 distribution. The changes in junctional permeability of cells correlated with changes in the distribution of immunoreactive Cx43. Cells treated for 2 hr with 10 m monensin exhibited a reduced communication rate which was accompanied by increased vesicular cytoplasmic Cx43 staining and reduced punctate surface staining of junctional plaques. Addition of 1.0 mm 8-bromo-cAMP to these cultures had no effect on the rate of communication or the distribution of Cx43 compared to cultures treated with monensin alone. These data suggest that an effect of cyclic AMP on Cx43 gap junctions is to promote increases in gap junctional permeability by increasing trafficking and/or assembly of Cx43 to plasma membrane gap junctional plaques.We acknowledge the technical assistance of Richard Lewis and Meghan Abella. We thank Dr. Hugh Dookwah for contributions to the myometrial cell isolation protocol and Drs. Stephen H. Safe, Timothy D. Phillips, and Evelyn Tiffany-Castiglioni for helpful discussions. This work was funded by NIH (HD-26182, P42-ES04917, ES05871-01A1), the March of Dimes Birth Defects Foundation Basic Research grant #1-0796, and USDA 92-37203-7952.     

Preparation,characterization, and structure of half gap junctional layers split with urea and EGTA

Gap junctions, collections of membrane channels responsible for intercellular communication, contain two paired hemichannels (also called connexons). We have investigated conditions for splitting the membrane pair using urea. We have developed a protocol which consistently splits the gap junction samples with 60–90% efficiency. Our results indicate that hydrophobic forces are important in holding the two connexons together but that Ca²⁺ ions are also important in the assembly of the membrane pair. Greater yields and better structural integrity of split junctions were obtained with a starting preparation of gap junctions which had been detergent treated. Image analysis of edge views of single connexon layers reveal an asymmetry in the appearance of the cytoplasmic and extracellular surface. Cryo-electron microscopy and image analysis of split junctions show that the packing and structural detail of membranes containing arrays of single connexons are the same as for intact junctions, and that the urea treatment causes no gross structural changes in the connexon assembly.The antibodies used to analyze the protein composition in our samples were the generous gift of Dr. David Paul. We thank Dr. Camillo Peracchia for sending us a preprint of his chapter on molecular mechanisms of connexon gating and docking which helped guide our thinking about interconnexon forces and John Badger for information on divalent cation sites in protein structure. We thank Amani Thomas-Yusuf for technical assistance. The photographic expertise of Marie Craig is gratefully acknowledged. This work was funded by National Institutes of Health GM43217 to G.E.S. and GM18974 to D.A.G.     

Flight initiation byProstephanus truncatus in relation to time of day, temperature, relative humidity and starvation

Studies were carried out in the laboratory on the influences of time of day, temperature, relative humidity and starvation on flight initiation byProstephanus truncatus (Horn) (Coleoptera: Bostrichidae). Flight occurred throughout the 12 h photophase and at the beginning of the scotophase but peaked at 2–0 h before darkness. Temperature exerted a significant effect on flight. The frequency of flight take-off increased with temperature over the range 20–30°C but declined sharply at 35°C. Flight activity increased with starvation up to a maximum at 2 days after which it began to decline.