Whole-embryo culture of Drosophila : development of embryonic tissues in vitro

Summary We have devised techniques to culture whole, dissected embryos of Drosophila melanogaster. We examine multiple aspects of the morphological and physiological development of the epidermis, musculature, nervous system, and internal organs in this cultured preparation, and show that in vitro development closely parallels normal embryogenesis. These techniques permit a wide range of experimental manipulations during embryogenesis and allow us to extend observations through late embryonic stages, after cuticle deposition. Applications of this technique are presented.     

Abscisic acid biosynthesis during corn embryo development

In this study we examined the biosynthesis of abscisic acid (ABA) by developing corn (Zea mays L.) embryos. Three comparisons were made: ABA biosynthesis in embryos isolated from kernels grown in vitro with those grown in the field; the developmental profile of ABA content with that of biosynthesis; and ABA biosynthesis in corn embryos lacking carotenoid precursors with ABA biosynthesis in normal embryos. Embryos were harvested at various times during seed development and divided into two groups. Endogenous levels of ABA were measured in one group of embryos and ABA biosynthetic capacity was measured in the other group. The ABA biosynthetic capacity was measured with and without tetcyclacis (an inhibitor of ABA degradation) in embryos from both field-grown and in-vitro-grown corn kernels. Reduced-carotenoid (either fluridone-treated or genetically viviparous) embryos were also included in the study. Corn kernels developing under field and in-vitro conditions differed from each other in their responses to tetcyclacis and in their profiles of ABA biosynthesis during development. Therefore, in-vitro kernel culture may not be an appropriate substitute for field conditions for studies of embryo development. The developmental profiles of endogenous ABA content differed from those of ABA biosynthesis in isolated embryos of both in-vitro-and field-grown kernels. This indicated that ABA levels in the developing embryos were determined by import from the maternal tissues available to the embryos rather than by in-situ biosynthesis. In embryos with reduced levels of carotenoids, either fluridone-treated or genetically viviparous embryos, ABA biosynthesis was low or nonexistent. This result is expected for the presence of an indirect pathway of ABA biosynthesis and in the absence of ABA precursors.Abbreviations ABA abscisic acid - DAP days after pollination     

Actin acetylation in Drosophila tissue culture cells

In a permanent cell line derived from Drosophila embryos, cytoplasmic actin is produced as an unstable precursor, which is subsequently converted to a stable form. This conversion results in a reduction in isoelectric point, with no apparent change in molecular weight. The conversion involves an enzymatic acetylation, and results in an insensitivity to aminopeptidase digestion, suggesting N-terminal blockage. Both the acetylated and unacetylated actins can participate in the assembly of F-actin, but with different efficiencies.This work was supported by a grant from the NIH (GM 22866).     

Nitrogen assimilation in field-grown winter wheat: Direct measurements of nitrate reduction in roots using15N

Summary Nitrate fertiliser labelled with¹⁵N was applied to a field grown crop of winter wheat. Uptake and assimilation of fertiliser nitrate was studied by monitoring the appearance of labelled nitrate and labelled amino acids in the xylem sap. Shortly after applying¹⁵N-nitrate to the soil about 30 per cent of recently absorbed¹⁵N was in the reduced form, indicating that roots of cereal crops can make a substantial contribution in reducing nitrate. Seasonal changes in crop growth andin vivo NRA are also described.     

Agroinfection and nucleotide sequence of cloned wheat dwarf virus DNA

Cloned DNA of the geminivirus wheat dwarf virus (WDV) was successfully used to infect seedling wheat plants. The clone was derived from circular double-stranded viral DNA isolated from naturally infected tissue. The initiation of infection was mediated by Agrobacterium tumefaciens using cloned dimeric WDV genomes in a binary Agrobacterium vector. The WDV DNA which comprised the infectious clone was sequenced and is compared with the published sequence of a Swedish isolate of the same virus. The results confirm that the single WDV genome component of 2.75 kb carries all the information necessary for production of viral symptoms, virus particles and viral double- and single-stranded DNA forms.     

Ein bakterielles Kulturfiltrat mit antiphytoviraler Aktivität an Gurke/Gurkenmosaik-Virus (cucumber mosaic virus) und Tabak/Tabakmosaik-Virus (tobacco mosaic virus): (Kurze Mitteilung)

Es wurde die Intra‐ und Interpopulationsvariation untersucht, die durch Adaptationsreaktionen der Individuen (Klone) der Gemeinen Quecke (Elytrigia repens (L.) DESV.) am Versuchsort zum Ausdruck kommt. Die Klone stammten aus Populationen verschiedener Höhenlagen vom Territorium der östlichen Bundesländer Deutschlands. Die Ergebnisse zeigen, daß sich die Populationen in der Variabilität der Enzymaktivität (saure Phosphatase, Peroxidase, Polyphenoloxidase) sowie in der Variabilität der Isoenzymmuster (Peroxidase, Esterase) unterscheiden. Die höchste Variabilität der genannten Parameter wurde für Populationen nachgewiesen, die aus einer Höhenlage von ca. 400 m ü. NN stammten. Ausgehend von 400 m ü. NN nimmt die Variabilität in Richtung größerer und niedrigerer Höhenlagen von 400 m hin ab. Die Variabilitätsunterschiede, die unter gleichen Bedingungen nachgewiesen wurden, weisen auf die Existenz eines Zusammenhanges zwischen den typischen Adaptationsreaktionen von Populationen und der Gesamtheit der Selektionsfaktoren des natürlichen Milieus hin, von dem sie stammten. Es gibt einen Zusammenhang zwischen der nachgewiesenen Variabilität und der Überlebensrate nach einer Adaptationsdauer von fünf Jahren.     

Tracing lake trophic history with a chironomid–total phosphorus inference model

1. In the absence of historical water chemistry data, predictive biological indicator groups preserved in lake sediments can be employed to reconstruct the history of lake eutrophication. Diatoms are well established in this role, but to augment diatom‐based inferences of nutrient status we investigate the potential use of chironomid midges (Insecta: Chironomidae). 2. Canonical correspondence analysis (CCA) of modern chironomid assemblages in surface sediments from 44 lakes in the English Midlands and Wales, U.K., shows that five environmental variables (total phosphorus (TP), bottom dissolved oxygen, maximum lake depth, Secchi depth and surface water temperature) make a statistically significant (P < 0.05) contribution to explaining the variance in the chironomid data, of which TP makes the largest contribution (29%). 3. The relationship is used to develop a series of weighted averaging (WA) and partial least squares (PLS), (WA‐PLS) models to infer log₁₀TP. The models are evaluated by leave‐one‐out (jack‐knifing) cross‐validation. The simplest minimal adequate model is provided by WA with unweighted inverse deshrinking of root mean square error of prediction (RMSEP_jack=0.34 and r²_jack=0.60). 4. Using this model, the trophic history of Betton Pool, Shropshire, U.K., is reconstructed from the mid‐19th century to the present day and the results from the chironomid‐TP model are compared with inferences from a diatom‐TP model ( 13 ). Both reconstructions suggest that there was a gradual rise in TP since 1850 AD until about 1974, followed by a more pronounced and rapid increase that has continued until the present. Inferred TP values from the WA chironomid inference model agree with diatom‐inferred values. 5. The study demonstrates that fossil chironomid assemblages can be used to investigate quantitatively the trophic history of lakes.     

Concentration dependence of the subunit association of oligomers and viruses and the modification of the latter by urea binding.

A theoretical model is presented that accounts for the facilitation of the pressure dissociation of R17 phage, and for the partial restoration of the concentration dependence of the dissociation, by the presence of subdenaturing concentrations of urea. As an indifferent osmolyte urea should promote the stability of the protein aggregates under pressure, and the decrease in pressure stability with urea concentration demonstrates that such indirect solvent effects are not significant for this case, and that the progressive destabilization is the result of direct protein-urea interactions. By acting as a "homogenizer" of the properties of the phage particles, urea addition converts the pressure-induced deterministic dissociation of the phage into a limited stochastic equilibrium. The model establishes the origin of the uniform progression from the stochastic equilibrium of dimers, to the temperature-dependent and partially concentration-dependent association of tetramers, to the fully deterministic equilibrium observed in many multimers and in the virus capsids.