Plastid polarity and meiotic spindle development in microsporogenesis ofSelaginella

Summary Microsporogenesis inSelaginella was studied by fluorescence light microscopy and transmission electron microscopy. As in other examples of monoplastidic meiosis the plastids are involved in determination of division polarity and organization of microtubules. However, there are important differences: (1) the meiotic spindle develops from a unique prophase microtubule system associated with two plastids rather than from a typical quadripolar microtubule system associated with four plastids; (2) the division axes for first and second meiotic division are established sequentially, whereas as in all other cases the poles of second division are established before those of first division; and (3) the plastids remain in close contact with the nucleus throughout meiotic prophase and provide clues to the early determination of spindle orientation. In early prophase the single plastid divides in the plane of the future division and the two daughter plastids rotate apart until they lie on opposite sides of the nucleus. The procytokinetic plate (PCP) forms in association with the two slender plastids; it consists of two spindle-shaped microtubule arrays focused on the plastid tips with a plate of vesicles at the equatorial region and a picket row of microtubules around one side of the nucleus. Second plastid division occurs just before metaphase and the daughter plastids remain together at the spindle poles during first meiotic division. The meiotic spindle develops from merger of the component arrays of the PCP and additional microtubules emanating from the pair of plastid tips located at the poles. After inframeiotic interphase the plastids migrate to tetrahedral arrangement where they serve as poles of second division.Abbreviations AMS axial microtubule system - FITC fluorescein isothiocyanate - MTOC microtubule organizing center - PCP procytokinetic plate - QMS quadripolar microtubule system - TEM transmission electron microscope (microscopy)     

Monoclonal antibodies that depress a specific subset of multiple components of the glutathione-induced response ofHydra

Summary Reduced glutathione evokes a feeding response, the tentacle-ball formation inHydra japonica. This response consists of at least 5 components (R1–R5). We raised 6 monoclonal antibodies (mAbs), each of which depressed a specific subset of these components, and we also examined the immunocytochemical localization of antigens with these mAbs at light microscopic level. The 2 mAbs that depressed R2 and R4 bound to the cnidocils of the desmoneme and the stenotele nematocytes; the 3 mAbs that depressed R5 bound to the apical surface adjacent to the cnidocils of the nematocytes; and the 2 mAbs that depressed R1 and R3 bound to the apical spot structures of unidentified cells in the ectoderm.Together with the specificity of the action of the mAbs on the behavioral response, the correspondence between the effects on the response and the structures visualized with these mAbs suggests that these structures include components of the receptor-effector system relevant to chemoreception.     

Consumption of leaf detritus by a stream shredder: Influence of tree species and nutrient status

Four species of riparian vegetation (alder, birch, willow and poplar) were fertilized with nitrogen, phosphorus, nitrogen + phosphorus, or no fertilizer (control). The resulting leaf detritus (leached but not microbially colonized) was offered to a stream shredder, Hydatophylax variabilis (Trichoptera: Limnephilidae). In one experiment, shredder consumption of leaf detritus from different nutrient treatments (within tree species) was compared, and in a second experiment, consumption of different tree species (within nutrient treatments) was compared. Larvae preferred leaf detritus from nitrogen + phosphorus treatments (except in poplar where nitrogen treatment was preferred). Alder was preferred over other tree species for all treatments. Chemical and physical analyses of leaf litter showed differences between tree species and nutrient treatments in nutrient content, tannins and leaf toughness. Leaf consumption by larvae was positively associated with nitrogen content and negatively associated with condensed tannin content. Species composition and nutrient status of riparian vegetation may strongly influence detrital food webs in streams.     

Spatial patterns and feeding of meiobenthic harpacticoid copepods in relation to resident microbial flora

Distributional and feeding relationships of harpacticoid copepods and their microbial prey were examined in a tidal channel at Great Sippewissett Marsh. A horizontal zonation of photosynthetic microorganisms was composed of: 1) a diatom area; 2) a purple sulfur bacterial (Thiocapsa sp.) area; and 3) a clear area. Four species of harpacticoid copepods were associated with given areas. Leptocaris brevicornis occurred in very high densities in the diatom area but in relatively low densities in other areas. Mesochra lilljeborgi occurred in significantly higher densities in the purple and clear areas.Feeding experiments, using resident microbial flora labelled with NaH¹⁴CO₃ and ³H-thymidine, were conducted to determine which foods are 1) ingested but simply pass through the gut and 2) ingested, and retained. These experiments indicated that L. brevicornis ingested diatoms and the heterotrophs associated with the diatoms, but only retained the heterotrophic portion. Microscopic examination indicated that diatoms were passed out intact in feces. Oscillatoria sp. (cyanobacterium) was not ingested. Mesochra lilljeborgi ingested Spirulina sp. (cyanobacterium), Thiocapsa sp., and the heterotrophs associated with Thiocapsa but only retained the Thiocapsa label.These data for harpacticoids suggest that spatial distributions of meiofauna may be closely coupled with microbial food organisms which they consume. Also, that while several microbial foods may be ingested, only certain microbes are digested and assimilated as a food resource, further indicating the complexity of feeding relationships among the meiofauna.     

Enhanced formation of mouse hybridomas without hat treatment in a serum-free medium

Summary A newly developed, serum-free medium (NYSF-404) selects for antibody-producing hybridomas after fusion of antigen-sensitized mouse spleen cells with myeloma cell lines P3-X63-Ag8-U1 (P3-U1), P3-X63-Ag8-6.5.3 (Ag8.653), or P3-NSI/1-Ag4-1 (NS-1). Without the need for hypoxanthine-aminopterinthymidine (HAT) selection of hybrid cells, frequency of hybridoma formation in medium NYSF-404 is higher (twice) than that in serum- and HAT-containing medium. Colonies developed upon limiting dilution in the presence of the mortal parent myeloma cells in medium NYSF-404 and pure culture of antibody-secreting cells could be subsequently established. The results suggest that fusions can be done in serum-free medium and that the clonal growth of hybridomas is dependent on factors produced by parent myeloma cells under serum-free culture conditions. Such factors seem deficient in serum- and HAT-containing medium or are masked by serum.     

Nippostrongylus brasiliensis: feeding activity in the mouse

Nippostrongylus brasiliensis incorporated the fluorescent dye, Rhodamine B, while feeding in vivo. Uptake of dye in both sexes of helminth increased linearly from 30 to 120 min after the hosts were given dye per os. Feeding by female N. brasiliensis significantly exceeded that of the male at 4 and 5 days postinfection. Feeding declined in older helminths of both sexes. The density of helminths had no effect on their incorporation of dye in vivo. Feeding in male- and female-only groups of worms was similar to that seen in populations of mixed sexes. Feeding by helminths decreased during the first 36 h of food deprivation in the host, but increased during subsequent fasting of the host. Both sexes of N. brasiliensis resumed feeding within 15 min after the fasted hosts were fed. Growth of N. brasiliensis increased linearly from 4 to 7 days postinfection, based on dry weight. Seven-day-old and older females were significantly heavier than males.     

Effects of Pratylenchus zeae and Quinisulcius acutus Alone and in Combination on Sorghum

Host-parasite relationships of Pratylenchus zeae and Quinisulcius acutus, alone or in combination, were studied on sorghum in the greenhouse and laboratory. Q. acutus at 1,000 or 5,000 nematodes per 15-cm-d pot and P. zeae at 500 nematodes per pot significantly suppressed plant height and fresh and oven dry shoot and root weights. A mixture of 1,000 Q. acutus and 500 P. zeae per pot resulted in greatest suppression of growth. Roots of plants inoculated with Q. acutus alone were reduced in number and size and showed lesions and discoloration. Reproduction of this nematode 42 days after inoculation was much greater in treatments of 100 or 1,000 than 5,000 nematodes. The population density of the two species at 6 weeks after inoculation was significantly less when combined than for each species alone. When the two species were combined, reproduction of P. zeae was greater than that of Q. acutus, but the final populations per gram of root weight were the same. Q. acutus fed ectoparasitically on epidermal cells of sorghum roots in the zone of elongation and differentiation when observed under in vitro conditions.     

Stylet penetration by the bayberry whitefly, as affected by leaf age in lemon, Citrus limon

Bayberry whitefly (Parabemisia myricae [Kuwana]) crawlers were placed on young and mature lemon leaves and were allowed 7–9 days to settle. Afterwards, the nymphs were fixed and sectioned in situ on the leaves and the area of leaf under each whitefly was examined at 1 000 x for stylet penetration. Both stylets and stylet tracks were readily visible in the sections. The path of penetration was mostly intercellular and the objective appeared to be the phloem. Passage of the stylets through the plant tissue did not cause detectable damage to most cells; however, damaged plant cells occasionally were noticed. Nymphs that had moulted during the 7–9 day settling period reached the phloem significantly more often than those that were still in their first instar. In each of the three replicates, penetration in the mature leaf occurred significantly less often than penetration in the young leaf (4% vs. 72%, p<0.01, ²). Penetration appears to be inhibited in the mature leaf either by the leaf cuticle or by factors detected by the nymphs after very shallow penetration into the leaf. The cuticle of mature leaves was much thicker than the cuticle of young leaves and may have been a barrier to stylet penetration.

---

Résumé Des larves de premier stade (avant la fixation) de l'aleurode, Parabemisia myricae (Kuwana), ont été placées, pour qu'elles se fixent, pendánt 2 à 9 jours, sur des feuilles jeunes ou mûres de citronnier. Des morceaux de feuilles avec des larves fixées ont été trempés dans 2% d'agar; ce procédé a permis de maintenir les larves à leurs sites de fixation finale. Des pièces d'agar contenant les morceaux de feuilles avec les larves ont ensuite été trempées dans de la paraffine, et sectionnées en séries de 10 . Il était nécessaire de maintenir une basse température pendant l'opération afin d'obtenir de bonnes sections des feuilles mûres durcies. Les stylets et leurs traces étaient faciles à voir dans les sections teintées aux safranins, et fast green. Presque toutes les traces des stylets étaient intercellulaires et leur destination semblaient être de phloème. En général, la plupart des cellules ne semblaient pas endommagées, les larves qui avaient mué pendant la période de 7 à 9 jours de fixation avaient atteint significantivement plus le phloeme que les larves du premier stade. Dans trois essais, la pénétration des feuilles mûres était significativement moins fréquente que celle des feuilles jeunes (p<0.01, ²).La pénétration dans des feuilles mûres semble être empêchée par la cuticule ou par des facteurs perçus par les nymphes après une pénétration superficielle. La cuticule des feuilles mûres était beaucoup plus épaisse que la cuticule des jeunes feuilles et pourrait donc représenter une barrière à la pénétration des stylets.

     

Aedes aegypti: model for blood finding strategy and prediction of parasite manipulation

Aedes aegypti mosquitoes salivate during intradermal probing of vertebrate prey before ingesting blood (Griffiths and Gordon 1952). Nonsalivating mosquitoes locate blood more slowly; this difference was ascribed to an anti-platelet activity found in the mosquito's saliva (Ribeiro et al. 1984). Mosquitoes infected with Plasmodium gallinaceum suffer pathology that specifically impairs saliva anti-hemostatic activity but without reducing volume of output (Rossignol et al. 1984). The complexity of the feeding apparatus of mosquitoes provides opportunity for a variety of strategies in which pathogens may produce specific lesions that enhance their transmission, but the variables that affect the duration of probing by mosquitoes have not been defined. We sought to resolve this complexity by identifying and quantifying relevant parameters of probing behavior. Mosquitoes thrust their mouthparts repeatedly through their host's skin while searching for blood. Female A. aegypti thrust at 7-sec intervals. If this search results in success, feeding ensues. Alternatively, the mosquito "desists," the mouthparts stylets are withdrawn, and the mosquito attempts to feed at another site. Even after previous desistance, the probability of finding blood remains undiminished. Functions for the probability of feeding success and desistance over time were derived using data from observations on 300 mosquitoes. The probability of feeding success was interpreted as being a function of the density of vessels in the skin, their geometric distribution, and the conditions locally affecting hemostasis. During each probe, the probability of desisting increased linearly with time, and after desisting once, mosquitoes tended to desist more rapidly. A model was developed incorporating Monte Carlo simulation which closely fit observed data. By changing values for the several parameters of the probability functions, we predicted modes in which parasites may manipulate their hosts to enhance transmission, both to and from the vector. In particular, parasite strategies in the vector would include induced salivary pathology; increased duration of probing thrusts; decreased desistance time; and inhibited phagoreception. Predicted parasite strategies in the reservoir host would include increased skin vascular volume and impaired host hemostasis. Our model supports the hypothesis of a mutualistic interaction of malaria and mosquitoes.