Oviducal Contribution to Alteration of the Vitelline Coat in the Frog, Rana japonica. An Electron Microscopic Study

The vitelline coat (VC) surrounding coelomic eggs of the frog, Rana japonica , comprises bundles of filaments running both parallel and perpendicular to the egg surface. The coat gives little or no staining reaction with PA-CrA-Silver methenamine. In contrast, in the VC of uterine eggs the filament bundles are less conspicuous. and the interstices between the filament bundles stain strongly for carbohydrate. This alteration occurs during passage of the eggs down the first 1/20 th of the oviduct, the pars recta. The epithelium of the p. recta contains secretory cells, which contain electron-dense granules distinct from those in the jelly-secreting cells in more caudal portions of the oviduct. Treatment of coelomic eggs with an extract of p. recta followed by exposure to a sperm suspension resulted in marked swelling and softening of the VC. These results indicate that the contents of the granules secreted from the epithelial cells in the p. recta are deposited in the VC to increase its susceptibility to a fertilizing sperm.     

Increase in DNA Content of Primary Leaves of Phaseolus vulgaris upon Decapitation

Changes in DNA content of bean (Phaseolus vulgaris) primaryleaves after decapitation were investigated. When apical budswere removed at 11 d, DNA content per leaf increased by about20% at 15 d and then decreased in parallel with the controls.The RNA and chlorophyll contents, fresh weight, and leaf areaexpressed on a single leaf basis changed in the same manneras the DNA content in response to decapitation. But when bothapical and lateral buds were removed, all these values continuedincreasing during the test period. Thus, growing lateral budsand apical buds have the same effect on the DNA change in primaryleaves as that due to ageing of the leaves. Cell number perleaf was not increased by any treatment, indicating that theobserved increase in the DNA content of primary leaves is ascribableto an increase in DNA per cell. Next, the whole shoots above the nodes of primary leaves wereremoved at various ages. The response of primary leaves to decapitationvaried according to their age. With age, they lost the abilityto increase their fresh weight, area, and chlorophyll contentbut not their DNA and RNA contents in response to decapitation.Decapitation stimulated chloroplast replication only withinthe period in which chloroplasts were replicating in controlleaves, but it induced chloroplast enlargement at any age. Therefore,the increase in DNA content after decapitation may be partiallydue to an increase in the amount of chloroplast DNA. When stems were heat-girdled above the nodes of the primaryleaves, these leaves showed responses similar to but smallerthan those to decapitation. The senescence of primary leavesseems to be controlled by the distribution of substances whichare transported from the roots.     

Experiments Pertaining to the Suppression of Accessory Sperm in Fertilized Newt Eggs*

In the physiologically polyspermic eggs of the newt, Cynops pyrrhogaster, a number of accessory sperm undergo pronuclear formation along with a concomitant DNA synthesis, but degenerate after zygote nucleus formation. When denuded eggs were divided into two halves at various post-fertilization stages, the andromerogons produced before zygote nucleus formation but not after that stage cleaved at a high frequency. The accessory sperm were unable to participate in the cleavage when they were located in the half of the egg which was connected with the diploid merogon by a cytoplasmic bridge higher than 100 μm in height. The removal of the egg nucleus or the retardation of early post-fertilization nuclear events by treatment with cycloheximide resulted in the induction of multipolar cleavage. Continuous exposure of the fertilized eggs to aphidicolin showed that in the appreciable absence of the DNA synthesis many eggs underwent a first cleavage cytokinesis of a mostly abortive type, but failed to initiate the following cytokinesis at all. Cytological examinations in association with these experiments suggest that the observed suppression of accessory sperm includes the inhibition of centriolar replication under the influence of the zygote nucleus, resulting in the failure of cytasters corporating with nuclear-independent activity of cortical cytoplasm.     

COMPLETE ABROGATION OF IMMUNE RESPONSE TO SKIN ALLOGRAFTS AND RABBIT ERYTHROCYTES IN THE EARLY THYMECTOMIZED XENOPUS

Larvae of the clawed toad, Xenopus laevis , were thymectomized at the N ieuwkoop -F aber stages 45–47 (4- to 7-day-old) and 51 (18-day-old), and raised beyond metamorphosis to study their immune reactivities. The toads thymectomized at stage 51 and the unoperated and sham-operated controls rejected allografted skins in 16–37 days. In contrast, the toads thymectomized at stages 45–47 did not reject the grafts at all, and there was no lymphoid invasion into the grafted area. The toads thymectomized at stage 51 produced antibodies against rabbit red blood cells (RRBC), as did the controls. However, those thymectomized at stages 45–47 did not produce specific antibodies. Electrophoresis on cellulose acetate membrane and immunoelectrophoretic analyses proved that in spite of the total lack of observed immune responses, the thymectomized toads do possess a certain level of immunoglobulins. It is concluded that the thymus exerts its decisive role in the establishment of immune competence when it is still in the initial state of differentiation. In relation to the immunoglobulins found in the early-thymectomized toads, a discussion was also offered of the presence of a thymus-independent humoral immune system.     

Comparing wing loading,flight muscle and lipid content in ant‐attended and non‐attended Tuberculatus aphid species

Although all Tuberculatus aphids possess wings, some species associated with ants exhibit extremely low levels of dispersal compared with those not associated with ants. Furthermore, phylogenetic interspecific comparisons find significantly higher wing loading (i.e. higher ratio of body volume to wing area) in ant‐attended species. This observation indicates that ant‐attended species may allocate more of their body resources to reproductive traits (i.e. embryos) rather than flight apparatus (i.e. wings, flight muscle and lipid). The present study focuses on two sympatric aphid species and aims to investigate the hypothesized trade‐off in resource investment between fecundity and the flight apparatus; specifically, the ant‐attended Tuberculatus quercicola (Matsumura) and non‐attended Tuberculatus paiki Hille Ris Lambers. Species differences are compared in: (i) morphology, (ii) embryo production, (iii) triacylglycerol levels and (iv) wing loading and flight muscle. The results show that T. quercicola has a larger body volume, higher fecundity and higher wing loading compared with T. paiki, which has a smaller, slender‐shaped body, lower fecundity and lower wing loading. No significant difference is found between the species with respect to the percentage of triacylglycerol content in dry body weight. The flight muscle development is significantly lower in T. quercicola than in T. paiki. These results indicate that the additive effect of higher wing loading and the lower amount of flight muscle development in T. quercicola may increase the physical difficulty of flight, and hence be responsible for its lower dispersal ability. The trade‐off between fecundity and dispersal documented in wing‐dimorphic insects may therefore be applicable to T. quercicola, which has fully developed wings.     

CO2‐mediated changes of plant traits and their effects on herbivores are determined by leaf age

1. Concentration of atmospheric CO₂ is predicted to double during the 21st century. However, quantitative effects of increased CO₂ levels on natural herbivore–plant interactions are still little understood. 2. In this study, we assess whether increased CO₂ quantitatively affects multiple defensive and nutritive traits in different leaf stages of cyanogenic wildtype lima bean plants (Phaseolus lunatus), and whether plant responses influence performance and choice behaviour of a natural insect herbivore, the Mexican bean beetle (Epilachna varivestis). 3. We cultivated lima bean plants in climate chambers at ambient, 500, 700, and 1000 ppm CO₂ and analysed cyanogenic precursor concentration (nitrogen‐based defence), total phenolics (carbon‐based defence), leaf mass per area (LMA; physical defence), and soluble proteins (nutritive parameter) of three defined leaf age groups. 4. In young leaves, cyanide concentration was the only parameter that quantitatively decreased in response to CO₂ treatments. In intermediate and mature leaves, cyanide and protein concentrations decreased while total phenolics and LMA increased. 5. Depending on leaf stage, CO₂‐mediated changes in leaf traits significantly affected larval performance and choice behaviour of adult beetles. We observed a complete shift from highest herbivore damage in mature leaves under natural CO₂ to highest damage of young leaves under elevated CO_2. Our study shows that leaf stage is an essential factor when considering CO₂‐mediated changes of plant defences against herbivores. Since in the long run preferred consumption of young leaves can strongly affect plant fitness, variable effects of elevated CO₂ on different leaf stages should receive highlighted attention in future research.     

Seasonal changes of phospholipids in last instar larvae of rice stem borer Chilo suppressalis Walker (Lepidoptera: Pyralidae)

Phospholipids of the cell membrane have been studied from the viewpoint of how overwintering insects inhabiting temperate zones adapt to low temperature. The transition of cell membrane phospholipids from a liquid crystalline phase to a gel phase is a crucial cause of cold or freezing injuries. We determined the qualitative and quantitative changes of phospholipids in the last instar larvae of the rice stem borer in summer and winter. We found that the total amount of their phospholipids did not change significantly between summer and winter and that the sum of phosphatidylcholine (PC) and phosphatidylethanolamine (PE) comprised about 85% of their total phospholipids. In summer, the ratio of their PE to PC was almost one, while from autumn to mid‐winter it increased and reached three in February. The fatty acid compositions of PC hardly changed, and the percentage of unsaturated fatty acids did not exceed 50%. In contrast, the percentage of unsaturated fatty acids of PE in overwintering larvae increased up to 80% as ambient temperatures fell and oleic acid mainly contributed to the high percentage of unsaturation. In the present study, the relationship between the quantitative and qualitative changes of phospholipids and adaptation of the rice stem borer to cold winter climate are discussed.