Arabidopsis thaliana ASN2 encoding asparagine synthetase is involved in the control of nitrogen assimilation and export during vegetative growth

We investigated the function of ASN2, one of the three genes encoding asparagine synthetase (EC 6.3.5.4), which is the most highly expressed in vegetative leaves of Arabidopsis thaliana. Expression of ASN2 and parallel higher asparagine content in darkness suggest that leaf metabolism involves ASN2 for asparagine synthesis. In asn2‐1 knockout and asn2‐2 knockdown lines, ASN2 disruption caused a defective growth phenotype and ammonium accumulation. The asn2 mutant leaves displayed a depleted asparagine and an accumulation of alanine, GABA, pyruvate and fumarate, indicating an alanine formation from pyruvate through the GABA shunt to consume excess ammonium in the absence of asparagine synthesis. By contrast, asparagine did not contribute to photorespiratory nitrogen recycle as photosynthetic net CO₂ assimilation was not significantly different between lines under both 21 and 2% O₂. ASN2 was found in phloem companion cells by in situ hybridization and immunolocalization. Moreover, lack of asparagine in asn2 phloem sap and lowered ¹⁵N flux to sinks, accompanied by the delayed yellowing (senescence) of asn2 leaves, in the absence of asparagine support a specific role of asparagine in phloem loading and nitrogen reallocation. We conclude that ASN2 is essential for nitrogen assimilation, distribution and remobilization (via the phloem) within the plant.     

Divergent enhancer haplotype of ebony on inversion In(3R)Payne associated with pigmentation variation in a tropical population of Drosophila melanogaster

The pattern and intensity of pigmentation have direct impact on individual fitness through various ecological factors. In a Drosophila melanogaster population from southern Japan, thoracic trident pigmentation intensity of most of the strains could be classified into Dark or Light‐type. The expression level variation of the ebony gene correlated well with this phenotype and the allelic differences in expression indicated that the variation is partly due to cis‐regulatory changes. In the ～13 kb gene region, we identified 17 nucleotide sites and 2 indels that were in complete association with the thoracic trident pigmentation intensity. Interestingly, 11 out of 19 sites located within ～0.5 kb of the core epidermis enhancer. These sites had no obvious association with the abdominal pigmentation intensity in the previously analysed African populations from Uganda and Kenya, which suggested that multiple potential mutational pathways in the cis‐regulatory control region of a single gene could lead to similar phenotypic variation within this species. We also found that the Light‐type enhancer haplotype is strongly linked to a cosmopolitan inversion, In(3R)Payne, which is predominant in warmer climatic regions in both hemispheres. The sequence pattern suggested that the strong linkage may be due to selective forces related to thermal adaptation. The inferred selection for lighter pigmentation in the Japanese population is in the opposite direction of the previously reported case of selection for darker individuals in African populations. Nevertheless, both adaptive changes involved cis‐regulatory changes of ebony, which shows that this gene is likely to be a common target of natural selection.     

Diverse diet compositions among harpaline ground beetle species revealed by mixing model analyses of stable isotope ratios

1. Species diversities of some insect lineages have been attributed to differentiation of feeding habits among species. Our objective was to determine variation in diet composition among harpaline ground beetle species occurring in a riverside grassland. 2. We examined the diet compositions of 14 species from six genera in the spring and 10 species from two genera in the autumn. We performed measurements of nitrogen and carbon stable isotope ratios in consumers and in their potential food items, and estimated relative contributions of different food items with two mixing models, IsoSource and MixSIR. 3. IsoSource and MixSIR software gave similar results, but IsoSource tended to calculate higher contributions of principal food items and smaller percentile ranges than MixSIR. Among harparine beetle species, there were diverse food utilisation patterns among four food categories (detritivorous invertebrates, herbivorous invertebrates, C3 plants, and C4 plants). Detritivores comprised the main diets of abundant harpaline species in the spring, whereas abundant harpaline species in the autumn were primarily herbivores feeding on C4 plants, or omnivores feeding on herbivorous invertebrates and C3 plants. Seasonal changes in food use were related to seasonal changes in the abundance of each food resource. 4. Mixing model analysis of stable isotope ratios is a convenient and effective method for roughly estimating diets of many species with diverse food habits (such as ground beetles). This method can contribute to determining the trophic relationships of related insects in one ecosystem.     

Structure and Role of the Five Glycopeptides of Human IgM Immunoglobulins

Carbohydrate is attached at five sites in the constant sequence region of the µ heavy chain of human IgM. The oligosaccharides are of two kinds, simple and complex and affect the conformation and properties of macroglobulins.     

Cell Fusion and Its Application to Genetic Analysis of Development in Dictyostelium discoideum

Dictyostelium cells were fused by a modification of the polyethylene glycol method of Kuhn and Parish. In the modified method Tricine buffer and Concanavalin A were used in place of Ca⁺⁺. The efficiency of genetic complementation through cell fusion was about 10 times higer by the modified method than by the original method with glycine buffer and Ca⁺⁺. Complementation between developmental mutants without any selectable growth character was clearly detected by the modified system, at efficiencies of about 1 in 10–20 surviving cells.     

A simple method to score single nucleotide polymorphisms based on allele‐specific PCR and primer‐induced fragment‐length variation

We describe a simple protocol to genotype single nucleotide polymorphisms (SNPs), which combines allele‐specific polymerase chain reaction (PCR) with fragment‐length analysis. Three primers are used in the PCR: two allele‐specific forward primers with a length‐difference and one reverse primer. The forward primers induce a length‐difference between the SNP‐variants, which can be assessed with standard fragment‐length analyses. We designed primers for 21 SNPs, and codominance was achieved for 76% of these SNPs. An inexpensive and flexible laser‐detection scoring protocol can be achieved with multiplex scoring and by incorporating the M13(‐21) genotyping method.     

Carapace formation of the podocopid ostracode Semicytherura species (Crustacea: Ostracoda)

Details of ostracode carapace structures were examined by SEM and TEM. The podocopine ostracode Semicytherura kazahana has major ridges on the carapace surface and develops its prismatic layer inside the adult carapace. Electron microscopy at the final molt reveals that the major ridges arise from the highly dense formation of pits within the underlying swollen epidermis, and that disappearance of the epidermis in the presumptive area of the prismatic layer occurs after the calcification of the outer lamella cuticle, and just before synthesis of the membranous layer. These facts suggest that the formation of the carapace in Semicytherura takes place via a more complex process than that of the other podocopid ostracodes.     

Photoperiodic induction of diapause in the silkworm, Bombyx mori: location of the photoreceptor using a chemiluminescent paint

ABSTRACT. To locate the photoreceptor involved in the photoperiodic induction of diapause in Bombyx mori L., covering of larval head with black paint or local illumination using chemiluminescent paint was carried out. A silkworm race showing a response of long-day type during the larval stage was employed. The results demonstrated that the photoreceptor is located in the head but is extraocular. The optical properties of the larval body suggest that during the first and second stadia light is admitted through the translucent clypeus of the head, but during later stadia enters over the entire larval body including the head, and that it reaches the cerebral lobe where a photoreceptor is possibly located.