Linkage analysis of the myosin heavy chain gene in Dictyostelium discoideum using a mutation generated by homologous recombination

Summary A mutation (mhcA1 in strain HMM) created by insertional gene inactivation was used to map the Dictyostelium discoideum myosin heavy chain gene (mhcA) to linkage group IV. Three phenotypic traits associated with this mutation (slow colony growth, inability of the mutant to develop past aggregation, and the presence of five to ten integrated vector copies) cosegregated as expected for the consequences of a single insertional event. This linkage was confirmed using a restriction fragment length polymorphism. The mhcA1 mutation was recessive to wild type and was nonallelic with mutations at the following loci on linkage group IV: aggJ, aggL, couH, minA, phgB and tsgB. This work demonstrates the ability to apply standard techniques developed for D. discoideum parasexual genetic analyses to mutants generated by transformation, which is of particular relevance to analysis of genes for which no classical mutations or restriction fragment length polymorphisms are available.     

Substrate specifity of the hexose carrier in the plasmalemma of Chenopodium suspension cells probed by transmembrane exchange diffusion

Substrate specifity of the proton-driven hexose cotransport carrier in the plasmalemma of photoautotrophic suspension cells of Chenopodium rubrum L. has been studies through the short-term perturbation of ¹⁴C-labelled efflux of 3-O-methyl-d-glucose. Efflux, occurring exclusively via carrier-mediated exchange diffusion, is trans-stimulated by the substrate and trans-inhibited by the glucose-transport inhibitors phlorizin (K _1/2=7.9 mM) and its aglucon phloretin (K _1/2=84 μM); with both inhibitors, 3-O-methyl-d-glucose efflux may be blocked completely. Trans-stimulation of efflux (up to fourfold) by a variety of the d-enantiomers of neutral hexoses, including glucose (K _1/2=48 μM), 3-O-methyl-d-glucose (K _1/2=139 μM), and fructose (K _1/2=730 μM), but not by, for instance, d-allose, and l-sorbose, shows that carrier-substrate interaction critically involves the axial position at C-1 and C-3, respectively. We suggest that substrate binding by the Chenopodium hexose carrier involves both hydrophobic interaction with the pyran-ring and hydrogen-ion bonding at C-1 and C-3 of the d-glucose conformation.     

Evolutionary trends of the hairy-faceSaguinus in terms of the dental and cranial morphology

Shape variations in the dentition and the cranium were analyzed for sevenSaguinus forms of the hairy-face tamarin by applying the factor analysis method. The results obtained for the dental and cranial measurements were almost consistent with each other. The magnitude of the difference in shape factors between theS. nigricollis group and theS. midas group is appreciably larger than that between the former group and theS. mystax group. If the ancestral geographic centre of origin is postulated as being within the region which is inhabited by the livingS. nigricollis group, the morphological distances between any pairs of groups correlate well with the geographic distances between them. Concerning the dental and cranial morphologies, the physical changes in the three species group probably took place in two directions; that is, from theS. nigricollis group to theS. mystax group, and from theS. nigricollis group to theS. midas group. The forms belonging to each species group are more closely related to each other, with the exception ofS. imperator in theS. mystax group. The uniqueness ofS. imperator was clearly demonstrated by factor analysis and distance analysis. In theS. mystax group, although still hypothetical,S. imperator may have been related only through the basic ancestral stock toS. labiatus andS. mystax.     

Effects of phorbol esters on endothelial cell microfilaments: Laser scanning confocal microscopy and quantitative morphometry of dose dependent changes

Phorbol esters are known to alter microfilaments but it is not clear if the changes correspond to modulation of the phosphoinositide turnover/protein kinase C system. The novel technique of laser scanning confocal epifluorescence was used to study fiber orientation in phorbol ester treated cells. We treated endothelial cells with control agents and agents known to stimulate protein kinase C: 4 alpha-phorbol, phorbol 12-myristate 13-acetate (PMA), phorbol dibutyrate (PDB), or lipopolysaccharide. After incubation with the test agents, the endothelial cell microfilaments were stained with rhodamine pholloidin and viewed by conventional epifluorescence and by laser scanning confocal epifluorescence microscopy. The images obtained by the confocal microscopy corresponded to a thin optical section through the cells, 300 nm or more in thickness. The microfilaments extended predominantly in the plane of focus. After exposure of the cells to phorbol esters, the stress fibers became more nearly parallel in arrangement or were shortened, but remained in the plane of focus. The modification of microfilaments in response to phorbol esters was quantitated by a single blind analysis. In order to compare the morphological changes with a biochemical action of the phorbol esters, we measured phosphoinositide turnover. The dose-dependence of morphological changes was compared and contrasted to the dose-dependent effect of phorbol esters on bradykinin-stimulated phosphoinositide turnover. PMA had about the same EC50 (1-5 nM) for both biochemical and morphological processes. PDB was less potent in inducing the disruption of microfilament structure than in inhibiting phosphoinositide turnover. Lipopolysaccharide was ineffective in inducing a morphological change under these conditions. A simple activation of protein kinase C is insufficient to explain the dose-dependent effects of phorbol esters. Thus a morphometric analysis can help distinguish the potency of cytoskeleton modulators.     

Limited chloroplast gene transfer via recombination overcomes plastomegenome incompatibility between Nicotiana tabacum and Solanum tuberosum

Green cybrids with a new nucleus-chloroplast combination cannot be selected after protoplast fusion in the intersubfamilial Nicotiana-Solanum combination. As an approach to overcome the supposed plastomegenome incompatibility, a partial plastome transfer by genetic recombination has been considered. After fusions of protoplasts of a light-sensitive Nicotiana tabacum (tobacco) plastome mutant and lethally irradiated protoplasts of wild-type Solanum tuberosum (potato), a single green colony was recovered among 2.5×10⁴ colonies. The regenerated plants had tobacco-like (although abnormal) morphology, but were normally green, and sensitive to tentoxin, demonstrating chloroplast markers of the potato parent. Restriction enzyme analysis of the chloroplast DNA (cpDNA) revealed recombinant, nonparental patterns. A comparison with physical maps of the parental cpDNA demonstrated the presence of a considerable part of the potato plastome flanked by tobacco-specific regions. This potacco plastome proved to be stable in backcross and backfusion experiments, and normally functional in the presence solely of N. tabacum nucleus.     

Value of a control decision rule for leek moth infestations in leek

Fourteen commercial leek fields with first and second generation Leek moth, Acrolepiopsis assectella Z., (LM) injury were sampled during 1986 in The Netherlands. For both generations, plant injury was more prevalent in the perimeter of the field than in interior portions. A sequential sampling program for use in making treatment decisions for LM was developed. An economic analysis of the value of sampling information derived from the sequential sampling program was performed. The parameters used for the analysis were crop yield and value, expected level of LM infestation, potential loss of value due to LM infestation, effectiveness of insecticide application, and cost of sampling. Due mainly to the high value of the crop and low cost of treatment, analysis indicates that there is little difference between a sampling-based management plan and prophylactic application of insecticides in terms of pest control costs. Additionally, such a sampling-based management plan is relatively insensitive to changes in the parameters used in the model. Thus, development of a threshold linked to a sampling procedure will not reduce pest control costs. Use of the sampling-based management plan will also not significantly increase pest control costs and will likely result in reduced insecticide use compared with a prophylactic treatment program.

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Zusammenfassung In den Niederlanden wurde 1986 der durch den Befall der ersten und zweiten Generation der Lauchmotte (Acrolepiosis assectella Z.) verursachte Schaden auf 14 kommerziell genutzten Porreefeldern ermittelt. Für beide Generationen wurde an den Feldrändern ein deutlich höherer Befall als in dem inneren Bereich eines Feldes festgestellt. Zur Bekämpfung der Lauchmotte wurde als Entscheidungsmodell ein sequentielles Probenahmeverfahren entwickelt. Der Informationsgehalt der sequentiellen Befallserhebung wurde einer ökonomischen Bewertung unterzogen. Hierbei wurden folgende Parameter zugrunde gelegt: Erntemenge und Ertrag, erwarteter Befallsgrad sowie hierdurch verursachter möglicher Ertragsverlust, Wirkungsgrad einer Insektizidbehandlung und Kosten der Stichprobennahme. Die Analyse der einzelnen Parameter ergab nur eine geringe Kostendifferenz zwischen einer gezielten Bekämpfung nach sequentieller Probenahme und prophylaktischen Insektizidspritzungen, bedingt durch den hohen Wert der Kultur und die niedrigen Kosten einer Bekämpfung. Darüber hinaus its das Verfahren einer gezielten Lauchmottenbekämpfung gegeüber Veränderungen der obengenannten Parameter relativ unempfindlich. Deshalb wird die Entwicklung eines Schwellenwertes der mit Probenahmeverfahren verbunden ist die Bekämpfungskosten nicht reduzieren. Die anwendung der Probenahmeplans würde Bekämpfungskosten auch nicht erhöhen und würde wahrscheinlich Insecktizidverbrauch reduzieren im Vergleich zu einen prophylaktischen Behandlungsprogramm.

     

Microscopic observations on the interaction of the mycoparasite Verticillium biguttatum with Rhizoctonia solani and other soil-borne fungi

The mycoparasitic interactions of Verticillium biguttatum with Rhizoctonia solani and with a variety of other soil-borne fungi were investigated in dual cultures. V. biguttatum interacted with various soil fungi by appressed growth along the host hyphae and infrequent penetrations. Intracellular growth and subsequent sporulation, however, only occurred with R. solani, a few binucleate Rhizoctonia and Ceratobasidium spp., and Sclerotinia sclerotiorum. Effective mycoparasitism on sclerotia was restricted to those belonging to R. solani.Electron-microscopic observations revealed that V. biguttatum can penetrate the host cell with infection tubes. This process is probably mediated by enzymatic hydrolysis of the cell wall. Subsequently, trophic hyphae develop within the host cytoplasm, ultimately resulting in death of the host cell.     

Test of Phenice's technique for determining sex from the os pubis

Pubic bone morphology was examined to test the accuracy of Phenice's visual method for determining sex from the os pubis. Twelve participants scored 50 pubic bones from individuals of known sex aged 52-92 years. The sample is of modern males and females, all presumed whites. An accuracy of approximately 83% in determining sex was recorded, compared to 95% reported by Phenice. This accuracy difference may reflect different age distributions of the two samples. Through replication of test results on two series of 25 specimens, the technique was found to be reliable. Previous experiences in human osteological analysis was shown to have no effect on accuracy in this test, confirming Phenice's assertion that the technique does not require extensive experience to yield accurate results. Results suggest that there is a moderate negative correlation between accuracy in determining the sex of an individual and that individual's age.