Phylogeography of the leaf beetle Chrysolina virgata in wetlands of Japan inferred from the distribution of mitochondrial haplotypes

The genetic differentiation among populations of the leaf beetle Chrysolina virgata living in wetlands of Japan was studied based on the sequence data of the mitochondrial cytochrome oxidase subunit I gene region (750 bp). Two distinct lineages of mitochondrial haplotypes were found: one (clade A) consisted of 26 haplotypes distributed over the distribution range of C. virgata between north‐east Honshu and Kyushu, whereas the other (clade B) was monotypic and confined to a small region in north‐east Honshu where it coexisted with clade A. Nested clade analysis for these haplotypes suggested that range expansion and following differentiation due to isolation by distance might have resulted in the present distribution pattern of the haplotypes in clade A. We discuss the evolutionary process leading to the occurrence of two distinct haplotype clades in Japan in terms of repeated colonization from the continent and range expansion and contraction during climatic changes.     

Contagious egg-distribution on host pods and avoidance of pods received eggs in a wild bruchidKytorhinus sharpianus

The present study investigated the way a wild bruchid, Kytorhinus sharpianus, distributes its eggs on host pods. First, we counted the number of eggs per pod on the host plant (Sophola flavescens) in the field. The egg-distributions by the overwintered-generation adults in July and by the second-generation adults in mid October did not deviate from Poisson distribution because of small numbers of eggs laid. However, the egg-distribution in late August when many first-generation adults emerged was significantly contagious. The deposition of eggs was determined for three types of pods: (A) clean pods, (B) pods with eggs, and (C) those which eggs had been laid on but were removed from later. The numbers of eggs laid newly on three types of pods were significantly different; A>C>B, suggesting that an oviposition marker pheromone (O.M.P.) caused females to avoid pods on which eggs were deposited. Egg-shells reinforced that response. Monte Carlo simulations for the egg-distribution on pods predicted that even if females avoid pods with heavy egg-load through O.M.P., the egg-distribution becomes contagious when there is a large heterogeneity in quality of the pods.     

Bi- and trivoltine complex life cycles in a Kanto (Japan) population of a wild bruchid Kytorhinus sharpianus

The partial trivoltinism and overwintering of Kytorhinus sharpianus Bridwell (Coleoptera: Bruchidae) was studied in the Kanto district, Japan. The later in the summer eggs were laid by the first-generation adults, the higher was the incidence of larval diapause in the second generation. The incidence of diapause also fluctuated between years, influencing the abundance of third-generation larvae. A relatively large proportion of third-generation larvae did not attain the diapause stage by the beginning of winter. The diapause development of larvae in diapause was completed by mid-January. Immature larvae of the third generation also overwintered and emerged as adults in the spring.     

Some properties and occurrence of cytochrome c-552 in the aerobic photosynthetic bacterium Roseobacter denitrificans

Characteristics and occurrence of cytochrome c-552 from an aerobic photosynthetic bacterium, Roseobacter denitrificans, were described.Relative molecular mass of the cytrochrome was 13.5 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and 15,000 by gel filtration. This cytochrome was a acidic protein having a pI of 5.6 and E_m was +215 mV at pH 7.0. Absorption peaks were at 278, 408 and 524 nm in the oxidized form and 416, 523 and 552 nm in the reduced form.Amino acid composition and N-terminal amino acid sequence of cytochrome c-552 determined for 24 residues had low similarities to those of cytochrome c-551 of this bacterium, which is homologous to cytochrome c ₂, although the physico-chemical properties of these two cytochromes were similar to each other.Cytochrome c-552 was maximally synthesized in the light under aerobic conditions but not in the dark. The synthesis also occurred in the presence of alternative acceptors such as trimethylamine N-oxide (TMAO) and nitrate under anaerobic conditions. Our results suggest that cytochrome c-552 is involved in TMAO respiration and denitrification in R. denitrificans, although the effect of light remains to be solved.Abbreviations E_m Midpoint redox potential - PAGE Polyacrylamide ge electrophoresis - SDS-PAGE Sodium dodecyl sulfate polyacrylamide gel electrophoresis - TMAO Trimethylamine N-oxide     

Induction of berberine biosynthesis by cytokinins in Thalictrum minus cell suspension cultures

Production of berberine could be induced by adding 6-benzylaminopurine (BAP) to Thalictrum minus cells, cultured in suspension in a medium containing 2,4-dichlorophenoxyacetic acid (2,4-D), early in the growth cycle. In the presence of BAP, the precursor, L-tyrosine, was rapidly converted into berberine which was then released into the medium, whereas substantial amounts of the intermediates, tyramine and dopamine, accumulated in non-berberine-producing cells grown in the same 2,4-D-containing medium without BAP. These results suggest that BAP activated enzymatic reactions subsequent to the formation of the amines in the biosynthesis of berberine.Abbreviations 2,4-D 2,4-dichlorophenoxyacetic acid - BAP 6-benzylaminopurine - NAA 1-naphthaleneacetic acid - IAP 6-isopentenylaminopurine - LS medium Linsmaier-Skoog medium - Growth medium LS medium containing 10^-6 M 2,4-D     

Monolignol and dilignol glycosides from Pinus contorta leaves

Four monolignol glucosides have been isolated and identified from the needles of Pinus contorta. Chavicol 4-O-α-L-arabinofuranosyl-(1 →     

Differential inhibition of dehydrogenase and 5-ene→4-ene isomerase activities of purified 3β-hydroxysteroid dehydrogenase. Evidence for two distinct sites

The success in synthesis of [³H]5-androstene-3,17-dione, the intermediate product in the transformation of DHEA to 4-androstenedione by 3β-hydroxysteroid dehydrogenase/ 5-ene→4-ene isomerase (3β-HSD) offers the opportunity to determine whether or not the two activities reside in one active site or in two closely related active sites. The finding that N,N-dimethyl-4-methyl-3-oxo-4-aza-5-androstane-17β-carboxamide (4-MA) inhibits competitively and specifically the dehydrogenase activity whereas a non-competitive inhibition type with a K_i value 1000 fold higher was observed for the isomerase activity, indicated that dehydrogenase and isomerase activities belong to separate sites. Using 5-dihydro-testosterone and 5-androstane-3β,17β-diol, exclusive substrates for dehydrogenase activity, it was shown that dehydrogenase is reversible and strongly inhibited by 4-MA and that thus the irreversible step in the transformation of DHEA to 4-androstenedione is due to the isomerase activity.