Stage-specific cuticular proteins of Ostertagia circumcincta and Ostertagia ostertagi

In this study we have shown that NHS-biotin and I¹²⁵-streptavidin can detect cuticular polypeptides of Ostertagia spp. The labelled polypeptide profile of intact nematodes is simple compared to the profile obtained by labelling homogenates. None of the major internal polypeptides are labelled and the subset of proteins labelled in intact nematodes appears to be mainly surface associated. The results presented here demonstrate that NHS-biotin may be used as a reagent for the analysis of surface polypeptides. The surface polypeptide profiles of the five major developmental stages (L1, L2, L3, L4 and adult) of Ostertagia circumcincta show a series of stage-specific molecules with no polypeptides common to all stages, indicating that the cuticle is a dynamic structure which changes throughout the life cycle. Similarity comparison of Ostertagia ostertagi L3 and L4 stage surface profiles showed that each stage is clearly distinct; comparison of these stages between the two species shows an overall similarity.     

The role of leaf surface wetness in larval behaviour of the sorghum shoot fly, Atherigona soccata

The susceptibility of sorghum to the shoot fly Atherigona soccata Rondani, (Diptera: Muscidae) is affected by seedling age and is highest when seedlings are 8–12 days old. This corresponds with high moisture accumulation on the central leaf which is the path of newly hatched larva as it moves downwards from the oviposition site, towards the growing apex. Studies showed that leaf surface wetness (LSW) of the central shoot leaf was higher in 10-day old seedlings than in seedlings of other ages. Similarly, LSW was much higher in the susceptible sorghum genotype CSH 1 than in the resistant genotype IS 2146. Larvae moved faster towards the growing point and produced deadhearts much earlier in CSH 1 than in IS 2146. They also moved faster in 10-day old seedlings than in seedlings of other ages. It was also shown that the leaf surface wetness of the central shoot leaf is a more reliable parameter of resistance than the glossy leaf trait or trichome density.

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L'influence de la humidité de la surface foliaire sur le comportement de la mouche des pousses du sorgho

Résumé La sensibilité du sorgho à la mouche des pousses du sorgho, Atherigona soccata Rondani, est liée à l'âge de la plantule. Elle est plus forte lorsque la plantule est âgée de 8 à 12 jours et la sensibilité est maximale à 10 jours. A ce stade de croissance on observe une forte accumulation d'humidité sur la feuille centrale de la tige. Les jeunes larves traversent cette zone humide lorsqu'elles descendent vers la zone de croissance à partir des pontes déposées sur la face ventrale des feuilles déroulées.Des études ont été menées à l'ICRISAT (Inde) sur la relation entre l'humidité de la feuille centrale de la tige des plantules du sorgho et les dégâts provoqués par la mouche des pousses. L'humidité de la surface des feuilles (HSF) a été estimée grâce à une échelle visuelle graduée 1 à 5 où, 1 = pas d'humidité apparente et 5 = surface de la feuille recouverte de gouttes d'eau. La HSF est plus élevée sur des pousses de sorgho âgées de 10 j que sur les pousses appartenant à d'autres classes d'âge. Les valeurs observées sont également plus fortes pour les variétés non résistantes à ce ravageur (CSH 1,4.8) que pour les variétés résistantes (IS 2146, (2)). La vitesse du déplacement larvaire entre le cornet et la zone de la croissance varie en fonction de l'âge de la plante et des cultivars. Les larves migrent plus rapidement vers la zone de croissance et provoquent la mort du coeur du sorgho plus tôt dans la variété CSH 1 que dans IS 2146. Les larves se déplacent plus rapidement dans les pousses âgées de 10 j que dans les pousses appartenant à d'autres classes d'âge.Des études ont également démontré que la HSF n'est pas directement liée au caractère feuille lisse où à la densité des trichomes. La HSF est faible pour les génotypes résistants présentent où non le caractère feuille lisse. Par contre la HSF est élevée pour les génotypes non résistants présentant le caractère feuille lisse ou non. Aucune relation directe entre la densité des trichomes et les dégâts provoqués par la mouche des pousses n'a pu être mise en évidence. L'analyse des correlations établie pour les caractères de surface des feuilles avec la mort du cur des sorghos indique que les correlations sont faibles et non-significatives pour le caractère feuille lisse (0.49) et la densité des trichomes (0.39 et 0.2). Par contre les correlations sont fortes et significatives pour la HSF (0.82).On conclue que la HSF de la feuille centrale de la tige est un facteur important dans le déterminisme de la résistance du sorgho vis à vis de la mouche des pousses. Les relations entre les processus physiologiques de la plante et les facteurs impliquées dans l'accumulation d'eau sur la surface des feuilles font actuellement l'objet d'études détaillées.

     

Wood excavations of three species of subterranean termites

We compared the feeding excavations on wood blocks of three species of subterranean termites, Coptotermes formosanus Shiraki, Reticulitermes flavipes (Kollar), and R. virginicus (Banks). Feeding rate followed the order C. formosanus > R. flavipes > R. virginicus. Wood surface area (mm²) exposed per unit feeding was higher for C. formosanus and R. flavipes than for R. virginicus. This was caused by the tendency of C. formosanus and R. flavipes to make internally penetrating tunnels, thereby increasing surface area, whereas R. virginicus made trough- and bowl-like depressions on the outside of blocks, sometimes decreasing the size of blocks outwardly without a corresponding high increase in surface area typical with the tunnels of the other species. Consequently, wood surface area was sometimes reduced, rather than increased as a result of feeding by R. virginicus. Different patterns of wood excavation suggest that these termites have divergent roles in wood decay processes.

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Résumé Les organismes pionniers qui modifient le bois et le rendent acceptable par les insectes qui le perforent sont généralement des champignons du bois pourri. Cependant, une fois que les termites ou autres insectes perforant le bois ont pénétré, leurs galeries favorisent les bactéries fixatrices d'azote, permettent l'invasion d'autres organismes décomposeurs, et de ce fait régularisent la décomposition du bois (Ausmus, 1977). L'exposition de la surface à l'intérieur des perforations jouant un rôle très important dans le processus de pourrissement, il est souhaitable de pouvoir quantifier la surface des galeries dues à l'alimentation des termites. Une courbe type permettant de prédire l'aire de la surface perforée a été construite en perçant 109 morceaux de bois de trous cylindriques de différents diamètres, en calculant l'aire de la surface des morceaux de bois, en appliquant et pesant une couche de vernis pour bois au polyuréthane, et en divisant la masse de polyuréthane par l'aire de la surface. Le modèle prédictif qui en découle est: Y=0,01443×-3,51825 (P=0,0001; r=0,68), y étant la masse de polyuréthane (en g) et x la surface (en mm²) du morceau de bois. En traitant de la même façon au polyuréthane les morceaux de bois perforés par les termites, nous pourrions déduire leur surface.Une expérience a été effectuée avec 3 espèces de rhinotermitides,- Coptotermes formosanus Shiraki, Reticulitermes flavipes (Kollar) et R. virginicus (Banks). Des groupes de chaque espèce se sont alimentés pendant 11 ou 12 jours sur des morceaux de bois non contaminés par des champignons. Nous avons déterminé la survie, la consommation, la modification de la surface du morceau de bois (par utilisation du modèle prédictif) et le changement de surface par terminte.La survie est la même, mais la consommation est dans l'ordre suivant: C. formosanus > R. flavipes > R. virginicus. L'aire de la surface exposée par unité d'alimentation était plus élevée pour C. formosanus et R. flavipes que pour R. virginicus (Tab. 1). Ceci est dû à la tendance de C. formosanus et R. flavipes de creuser des galeries vers l'intérieur, tandis que R. virginicus fait des cuvettes à la surface du bois. Les attaques superficielles de R. virginicus réduisent parfois le volume du morceau de bois sans accroître proportionnellement la surface comme le font les espèces creusant des galeries. Ainsi, avec R. virginicus la surface peut être réduite au lieu d'augmenter. Des différences entre colonies s'observent avec toutes les variables (Tab. 2).Nos résultats suggèrent que C. formosanus et R. flavipes contribuent plus que R. virginicus à exposer le bois aux autres organismes décomposeurs. Cependant, ces résultats peuvent être modifiés par un conditionnement préalable du bois par des champignons.

     

Early electrochemical events in lectin-lymphocyte interaction

Concanavalin A, at extremely low concentrations, will produce significant increases in the electrophoretic mobility of murine splenic T lymphocytes. It has been established that the alteration in cellular surface charge is mediated by a factor produced by those lymphocytes that have reacted directly with con A. We originally conjectured that the mobility change might be the consequence of an alteration in the distribution of the charged moieties of membrane glycoproteins. The results of experiments conducted at low temperature raise some questions about this mechanism. Further experiments have been performed to establish the nature of the physicochemical alterations in the peripheral zone of the factor-stimulated lymphocytes that are manifest as changes in cellular surface charge. The results of these studies indicate that, subsequent to the interaction of T lymphocytes with con A, there is a reduction in the number of positively charged amino groups effective at the electrophoretic surface of the cells.     

Isolation of Cell Surface Membranes from Cultured C6 Glioblastoma Cells

Abstract: Plasma membranes were isolated from C6 glioblastoma cells by two methods. In the first method cells were treated with concanavalin A and lysed in hypotonic medium. After partial separation of plasma membranes from other cell material, the lectin was displaced with a-methyl-D-mannoside. In the second method untreated cells or cells iodinated in a lactoperoxidase-catalyzed reaction were homogenized in isotonic medium. Membrane fractions obtaincd by either homogenization procedure were further purified by rate zonal and equilibrium centrifugations into linear density gradients. Disruption of the glioblastoma cell membrane gives rise to heterogeneous assemblies of mem- brane fragments. Two populations of plasma membranes were isolated from untreated and from iodinated cells: a "lighter")membrane fraction characterized by relatively lower sedimentation velocity and buoyant density, and a "heavier" membrane fraction of relatively faster sedimentation velocity and higher buoyant density. Both fractions showed electrophoretic patterns similar to those of ¹²⁵I-labeled cell surface proteins. Their specific (Na⁺+ K⁺)-ATPase activity was seven- to eightfold the homogenate activity (recovery, 13.1%). Both fractions were, however, still contaminated by smooth endo- plasmic reticulum, as judged from the activity 0: NADPH-dependent cytochrome c reductase (recovery, 2.4%). It is suggested that plasma membrane fragments present in the two fractions might differ in the organization of their structures, e.g., membrane vesicle intactness and membrane orientation.     

The mechanism of voltage-sensitive dye responses on sarcoplasmic reticulum

Summary The mechanism of voltage-sensitive dye responses was analyzed on sarcoplasmic reticulum vesicles to assess the changes in membrane potential related to Ca²⁺ transport. The absorbance and fluorescence responses of 3,3-diethyl-2,2-thiadicarbocyanine, 3,3-dimethyl-2,2-indodicarbocyanine and oxonol VI during ATP-dependent Ca²⁺ transport are influenced by the effect of accumulated Ca²⁺ upon the surface potential of the vesicle membrane. These observations place definite limitations on the use of these probes as indicators of ion-diffusion potential in processes which involve large fluctuations in free Ca²⁺ concentrations. Nile Blue A appeared to produce the cleanest optical signal to negative transmembrane potential, with least direct interference from Ca²⁺, encouraging the use of Nile Blue A for measurement of the membrane potential of sarcoplasmic reticulumin vivo andin vitro. 1,3-dibutylbarbituric acid (5)-1-(p-sulfophenyl)-3 methyl, 5-pyrazolone pentamethinoxonol (WW 781) gave no optical response during ATP-induced Ca²⁺ transport and responded primarily to changes in surface potential on the same side of the membrane where the dye was applied. Binding of these probes to the membrane plays a major role in the optical response to potential, and changes in surface potential influence the optical response by regulating the amount of membrane-bound dye. The observations are consistent with the electrogenic nature of ATP-dependent Ca²⁺ transport and indicate the generation of about 10 mV inside-positive membrane potential during the initial phase of Ca²⁺ translocation. The potential generated during Ca²⁺ transport is rapidly dissipated by passive ion fluxes across the membrane.     

Nutritional requirements for conidial germination and hyphal growth of Beauveria bassiana

Nutritional requirements for germination and growth of the entomopathogenic fungus Beauveria bassiana are not complex. For germination to occur, a utilizable source of carbon must be present; however, a nitrogen source is needed for continued hyphal growth, otherwise lysis ensues. Compounds that can serve as utilizable carbon-energy sources for germination include glucose, N-acetylglucosamine, glucosamine, chitin, starch, lanolin, hydrocarbons in crude oil, and some longer-chain fatty acids. Both organic and inorganic sources of nitrogen are readily utilized for growth. Conidia undergo active metabolism soon after being placed in a suitable growth medium, indicating that conidia are released from their state of dormancy several hours before emergence of the germ tube can be observed. Because of the nutritional versatility of B. bassiana, this fungus should be able to survive and be infective in several types of natural environments.     

Plate-Induced tumors of BALB/3T3 cells exhibiting foci of differentiation into pericytes,chondrocytes, and fibroblasts

The BALB/3T3 clone A31 mouse embryo cell line has been used by many investigators as a model “normal” “fibroblast” line for a variety of in vitro studies. It has been shown, however, that these cells are not “normal” because they will produce tumors within 2–4 months if 3 × 10⁴ cells are implanted subcutaneously in BALB/c mice attached to 0.2 × 5 × 10-mm plastic plates. Previous studies also suggested that these cells were not fibroblasts because they gave rise to tumors with the characteristics of vascular endothelium not fibroblasts. We now report that BALB/3T3 (clone A31), BALB/3T3-T, a proadipocyte subclone of clone A31 cells, and six recent subclones of BALB/3T3-T cells show additional differentiation patterns when tumors derived by implantation of these cells attached to plastic plates are examined. Differentiation into pericytes, chondrocytes, and fibroblasts was observed. We conclude that the BALB/3T3 clone A31 cell line and related lines are multipotent mesenchymal cells which are capable of differentiation into a variety of cell types.     

The maximum leaf surface temperatures of the higher plants observed in the Inland Sea area

The maximum leaf surface temperatures (MLSTs) of 126 species of higher plants were measured by means of an infrared thermometer, in the Inland Sea area, southwest of Honshu-Island, Japan, where plants suffered from severe environmental conditions due to an abnormally small amount of precipitation during the summer of 1978. The MLSTs of plants in the summer of 1978 were greater than or equal to those of 1979, when the environmental conditions were not so severe. The MLST measured in this study was 50.4 C for a non-succulent plant (Liriodendron), and 53.1 C for a succulent plant (Agave). Plants with different life forms appeared to have different MLSTs. The average of the MLSTs of conifers deciduous trees, and evergreens were 36.4, 37.7, and 40.3 C, respectively. This order corresponds to the distribution of forests from high to low, latitudes. Also the MLSTs were higher for woody plants than for herbaceous plants. Relatively high leaf temperatures were observed for climbing plants, both herbaceous and woody. Plants with narrow leaves had lower leaf surface temperatures than those with borad leaves. Herbaceous dicotyledonous plants actively growing at the end of the summer of 1978, in full sun at Hiroshima Castle were exclusively those with relatively high leaf temperatures.     

The mechanism of bicarbonate assimilation by the polar leaves of Potamogeton and Elodea. CO2 concentrations at the leaf surface

Abstract. Photosynthetic utilization of HCO, in leaves of Poiamogeton and Elodea occurs at the lower leaf side, with subsequent OH∼ release at the upper side. It is accompanied by transport of cations, in the present experiment K +, across the leaf. The resulting pH and K+ concentration changes near the leaf surface were recorded with miniature electrodes. From the pH and K+ concentration the concentrations of the different inorganic carbon species were calculated and compared with photosynthetic O, production. HCO⁻₃ utilization is accompanied by a drastic increase in the free CO₂ concentration near the lower epidermis. Experiments with CO₂− and HCO₃⁻free solutions showed an oscillating acidification near the lower epidermis and alkalinization near the upper epidermis. It is concluded that the acidification results from the activity of light-dependent H+ pumps. The finding that an increase in pH at the upper side always coincided with a decrease at the lower in these experiments shows that the H+ pumps and the OH− extruding mechanism are coupled although occurring in different cell layers. Previously we have suggested that the first step in the process of photosynthetic HCO₃⁻ utilization is external conversion of HCO₃⁻" by acidification caused by light-dependent H+ pumps. The present results strongly support this hypothesis. Two possible pathways for the accompanying K + transport are discussed. The model presented here explains the known inhibiting effects of buffers and high pH on photosynlhetic HCO₃⁻ utilization.