Accumulation and Spatial Distribution of Poly(A)+RNA in Oocytes and Early Embryos of Drosophila melanogaster

We describe the accumulation and distribution of poly (A)⁺RNA during oogenesis and early embryogenesis as revealed by in situ hybridization with a radio-labeled poly (U) probe. The amount of poly (A)⁺RNA in nurse cell cytoplasm continuously increased untill mid-vitellogenic stage (st. 10), then decreased with the rapid increase of poly (A)⁺RNA in the oocyte (st. 11). The localization of poly (A)⁺RNA at stage 10 was in the anterior region of the oocyte, where it is connected by cytoplasmic bridge to the nurse cells. These observations indicate that most of the poly (A)⁺RNA synthesized in the nurse cells is transferred to the oocyte through the cytoplasmic bridges at stage 10–11. During the remainder of oogenesis (st. 11–14) and during preblastodermal embryogenesis, poly (A)⁺RNA was evenly distributed over the cytoplasm of oocytes and embryos. At blastoderm stage, poly(A)⁺RNA became concentrated in the peripheral region of embryos. Though the somatic nuclei of the blastoderm contained a detectable amount of poly (A)⁺ RNA, the pole cell nuclei did not. The cytoplasmic RNA visualised by acridine orange staining and the poly (A)⁺RNA detected by hybridization with [³H]poly (U) exhibited identical distributions during oogenesis and early embryogenesis. These observations provide a basis to assess the unique distributions of specific RNA sequences involved in early development.     

Maternal Messenger RNA as a Determinant of Pole Cell Formation in Drosophila Embryos

Some polar plasm components are UV-sensitive. Messenger RNA extracted from oocytes or cleavage embryos can to induce pole cells in embryos that have been deprived of ability to form pole cells by UV-irradiation. This article reviews studies on the role of this mRNA in the developmental pathway leading to germ cell formation.     

Quantitative aspects of heterogeneity in soil organic matter dynamics in a cool-temperate Japanese beech forest: a radiocarbon-based approach

Soil is the largest carbon reservoir in terrestrial ecosystems; it stores twice as much carbon as the atmosphere. It is well documented that global warming can lead to accelerated microbial decomposition of soil organic carbon (SOC) and enhance the release of CO₂ from the soil to the atmosphere; however, the magnitude and timing of this effect remain highly uncertain due to a lack of quantitative data concerning the heterogeneity of SOC biodegradability. Therefore, we sought to identify SOC pools with respect to their specific mean residence times (MRTs), to use those SOC pools to partition soil respiration sources, and to estimate the potential response of the pools to warming. We collected surface soil and litter samples from a cool-temperate deciduous forest in Japan, chemically separated the samples into SOC fractions, estimated their MRTs based on radiocarbon (¹⁴C) isotope measurements, and used the data to construct a model representing the soil as a complex of six SOC pools with different MRT ranges. We estimate that a minor, fast-cycling SOC pool with an MRT of less than 10 years (corresponding to the O horizon and recognizable plant leaf fragments in the A1 horizon) is responsible for 73% of annual heterotrophic respiration and 44% of total soil respiration. However, the predicted response of these pools to warming demonstrates that the rate of SOC loss from the fast-cycling SOC pool diminishes quickly (within several decades) because of limited substrate availability. In contrast, warming will continue to accelerate SOC loss from slow-cycling pools with MRTs of 20–200 years over the next century. Although using a ¹⁴C-based approach has drawbacks, these estimates provide quantitative insights into the potential importance of slow-cycling SOC dynamics for the prediction of positive feedback to climate change.     

Herbivory in canopy gaps created by a typhoon varies by understory plant leaf phenology

1. Availabilities of light and soil nitrogen for understory plants vary by extent of canopy gap formation through typhoon disturbance. We predicted that variation in resource availability and herbivore abundance in canopy gaps would affect herbivory through variation in leaf traits among plant species. We studied six understory species that expand their leaves before or after canopy closure in deciduous forests. We measured the availabilities of light, soil nitrogen, soil water content, and herbivore abundance in 20 canopy gaps (28.3–607.6 m²) formed by a typhoon and in four undisturbed stands. We also measured leaf traits and herbivory on understory plants. 2. The availabilities of light and soil nitrogen increased with increasing gap size. However, soil water content did not. The abundance of herbivorous insects (such as Lepidoptera and Orthoptera) increased with increasing gap size. 3. Concentrations of condensed tannins, total phenolics, and nitrogen in leaves and the leaf mass per area increased in late leaf expansion species with increasing gap size, whereas none of the leaf traits varied by gap size in early leaf expansion species. 4. Herbivory increased on early leaf expansion species with increasing gap size, but decreased on late leaf expansion species. In these late leaf expansion species, total phenolics and C : N ratio had negative relationships with herbivory. 5. These results suggested that after typhoon disturbance, increased herbivory on early leaf expansion species can be explained by increased herbivore abundance, whereas decreased herbivory on late leaf expansion species can be explained by variation in leaf traits.     

INSTABILITY OF MESSENGER AND RIBOSOMAL RNA IN A GLUE-PROTEIN MUTANT OF Bombyx mori

The mucous glands of Bombyx pupae secrete glue proteins which attach deposited eggs to the mounting sheet. A mutant of a dominant gene, named no glue (Ng) , produces nonadhesive eggs which have a low capacity for glue-protein synthesis. In the present study it was shown that the mucous glands of Ng silkworms showed rapid degradation of mRNA as well as rRNA during development; this may cause the low capacity for glue-protein synthesis in the mutant organ. In contrast, the mucous glands of normal silkworms showed a significant increase in content of RNA's until the maximum rate of glue-protein synthesis was achieved. The degradation of RNA in the Ng mucous gland was inhibited by actinomycin D injected into the body fluid. Thus it is supposed that the Ng gene codes for a presumptive controller RNA, which would be the mediator of RNA instability in the mucous glands of Ng pupae.     

Tissue Distribution of an Immunogenic Melanoma Associated Antigen,D-1

A human melanoma-associated antigen immunogenic in patients was recently identified by screening an expression cDNA library constructed from cultured human melanoma cell line with sera from patients with melanoma. The nucleic acid sequence of the cloned D-1 cDNA has no significant homology with previously reported mammalian genes. The cDNA D-1 encodes a peptide of about 37 kDa, which showed fivefold higher reactivity with sera from patients with melanoma than with sera from normal donors. In order to detect D-1 gene expression in vivo, in-situ hybridization and immunostaining with cRNA probe and murine anti-D-1 sera were carried out on surgically removed tissues. Digoxigenin-labeled cRNA D-1 was exclusively hybridized with mRNA in the cytoplasm of melanoma cells but not with keratinocytes and fibroblasts adjacent to melanoma nests. Polyclonal anti-D-1 antibodies were obtained by immunization of Balb/c mice with recombinant D-1 peptide and clearly reacted with melanoma cells but not with keratinocytes and fibroblasts, similar to the results of in-situ hybridization. The above information will help to assess the suitability of recombinant D-1 peptide to implement active specific immunotherapy in patients with melanoma.     

Conditions for seedling establishment and probable function of seed in the clonal sedge Carex rugulosa Kük. in riverside marshes

Carex rugulosa Kük. forms large meadows in moist zones along estuarine riversides. These meadows are usually maintained by rhizomatous ramet production (clonal growth), and the appearance of seedlings is uncommon. We studied the conditions required for seedling establishment. In areas in which clonal ramets were dense (906–1050/m²), and the relative light intensity at ground level was low (0.8–3.8%), seed germination was entirely suppressed. In contrast, many seedlings (288–513/m²) were observed in areas in which clonal ramets had been considerably reduced (13–269/m²) by continuous inundation in the previous year. In these areas, the relative light intensity had increased (20.5–79.3%). It was inferred that seeds resisted the prolonged inundation that killed the ramets, and that germination was induced under these open conditions. These results suggest that the main functions of sexual reproduction in C. rugulosa are recovery after severe damage to vegetation using seeds from the seed bank and the dispersal of seeds to areas without dense vegetation. Most seedlings died when the marshy meadow was temporarily inundated by heavy rain. However, transplanted seedlings survived better at artificially elevated sites that were not submerged. This shows that inundation during the seedling stage impedes seedling survival. Therefore, seeds can contribute to recovery after ramet decline or colonization to open area when (i) the rainfall is not heavy to cause seedling submergence or (ii) seeds are dispersed to higher area which is not inundated.     

Longitudinal changes in δ13C of riffle macroinvertebrates from mountain to lowland sections of a gravel‐bed river

Summary 1. To understand longitudinal changes in the trophic base of benthic macroinvertebrates from mountain to lowland river sections, we investigated carbon stable isotopic compositions (δ¹³C) of macroinvertebrates and their food resources in riffles for four seasons at 14 sites along the main stem of the Toyo River, Japan. 2. At each site, δ¹³C was usually highest or nearly highest for periphyton (epilithic biofilm) and was lowest for transported leaf materials. Among macroinvertebrate groups, grazers usually had higher δ¹³C values than filterers or predators. 3. During all seasons, δ¹³C of periphyton and all macroinvertebrate groups increased downstream from mountain to upland sections, but decreased downstream from upland to lowland sections. In addition, the difference between grazer δ¹³C and filterer δ¹³C decreased from mountain to upland sections, but increased from upland to lowland sections. 4. The observed changes in δ¹³C of periphyton and macroinvertebrates from mountain to upland sections agree with previous reports: the δ¹³C of periphyton and consumers increased with stream size and productivity. The decrease in δ¹³C of periphyton and macroinvertebrates from upland to lowland sections has not been reported previously, and this may have resulted from an increased importance of terrestrial detritus relative to periphyton production in the lowland section, where riffles were infrequent and pools dominated the reach. 5. A simple mixing model of δ¹³C showed that grazers rely mostly on periphyton at all sites, whereas the importance of periphyton for filterers changed longitudinally increasing from mountain to upland sections and decreasing from upland to lowland sections. This longitudinal trend for filterers is possibly associated with the changes in the availability or quality of terrestrial detritus in transported particulate organic matter. 6. Longitudinal changes in the relative importance of autochthonous production and allochthonous detritus appear to be reflected in δ¹³C of riffle benthic communities. The longitudinal changes were not monotonic, and specific reach characteristics may be responsible for the greater importance of allochthonous detritus in mountain and lowland sections.     

Relationship between seed and clonal growth in the reproduction of Carex rugulosa Kük. in riverside meadows

Carex rugulosa Kük. develops riverside vegetation in brackish marshes along the lower parts of rivers. Because the rhizomatous ramets grow densely (590–950/m²) in its developed population, it is expected that seedlings cannot get enough light to grow if seeds germinate in crowded ramets. We studied the reproductive contribution of seed in a marshy meadow of the Ohashi River, Japan. The number of seeds produced was 6900–14 200/m² in 2002. The number of buried seeds in the following germination season differed among sites from 1190 to 2690/m². No seedlings were observed at plots where the ramets from rhizomes grew densely. In contrast, seedlings appeared in plots where all rhizomatous ramets were artificially cut. The number of seedlings corresponded to 17.5–39.5% of that of buried seeds. All these seedlings were submerged and died when it rained heavily. In the genotype analysis using allozyme detection, one of the phosphoglucoisomerase loci ( Pgi-2 ) indicated a pair of alleles. Among eight plots (each plot was 50 or 60-cm square), four showed an excess of heterozygotes and two showed homozygote domination in the Pgi-2 genotype. These results suggest that: (i) seed germination is suppressed in crowded ramets; (ii) seedling survival is severely reduced by inundation; and (iii) the population is usually maintained by clonal growth. As germination is induced in open areas, it is hypothesized that the main role of seed is recovery after vegetation decline caused by environmental stresses or colonization to other areas without dense vegetation.