Gas Chromatographic Determination of Optical Purity of a Chiral Derivatization Reagent,N-Acyl-l-prolyl Chloride

Two endo-type cellulases, tentatively called carboxymethyl cellulases (CMCases) I and II, were purified by gel filtration, ion-exchange chromatography, affinity chromatography, and chromato-focusing from a culture supernatant of Penicillium purpurogenum. Their homogeneity was verified by analytical polyacrylamide gel electrophoresis. The molecular weights of CMCases I and II, estimated by gel filtration, were 72,000 and 50,000, respectively. CMCases I and II contained about 12% and 8% carbohydrate, and had isoelectric points of 4.3 and 3.9, respectively. CMCase I produced cellobiose, glucose, and a trace amount of cellotriose from H₃PO₄-swollen cellulose and Avicel (microcrystalline cellulose), while CMCase II produced cellobiose and cellotriose with a small amount of glucose and cellotetraose. The products from reduced cellopentaose by both enzymes were released predominantly in the β-configuration. CMCase II appeared to act in more random fashion than I against carboxymethyl cellulose. These results suggest that both enzymes attack insoluble cellulose randomly, although there are some differences in the mode of hydrolytic action.     

cDNA cloning and gene expression of ascorbate oxidase in tobacco

A cDNA clone for ascorbate oxidase (AAO) has been isolated from a cDNA library of tobacco (Nicotiana tabacum) cells. The identity of the amino acid sequence deduced from tobacco AAO cDNA to that from pumpkin AAO cDNA was 68%, which was much lower than the identity (80%) between pumpkin and cucumber AAO. AAO activity in tobacco cells was much lower than that in pumpkin cells, whereas the immunoreactive protein in tobacco cells was more abundant than that in pumpkin cells. We suppose that AAO protein in tobacco cells may be less active than that in pumpkin cells. Genomic Southern blotting suggested that AAO in tobacco was encoded by a single-copy gene. Northern blotting revealed that mRNA of AAO was highly expressed in young and growing tissues of tobacco plant.     

Specific Secretion of Proline-Rich Proteins by Salt-Adapted Winged Bean Cells

Six proteins, designated SAP1 through SAP6, were secreted specificallyby salt-adapted cells of winged bean (Psophocarpus tetragonolobus)in suspension cultures. The amino-terminal amino acid sequencesof SAP2 (57 kDa), SAP4 (21 kDa), SAP5 (19 kDa) and SAP6 (17kDa) were homologous to the sequences of proline-rich proteins,indicating that proline-rich proteins are secreted specificallyby these salt-adapted cells. In addition, the amino-terminalamino acid sequence of SAP2 was identical to that of SAP4, andthe amino-terminal sequence of SAP5 was identical to that ofSAP6. Secretion of SAP2 was significantly enhanced by additionof AlCl₃ but not of KCl, LiCl, CaCl₂, MgCl₂, mannnitol or sucroseto suspension cultures. Furthermore, secretion of SAP4, SAP5and SAP6 was stimulated by addition of abscisic acid to cultures,suggesting that these proteins might be secreted in responseto salt or osmotic stress. (Received September 12, 1994; Accepted January 20, 1995)     

Analytical studies of light trap records in the Hokuriku district,II the green rice leafhopper,Nephotettix cincticeps

Light-trap records on the green rice leafhopper, Nephotettix cincticeps, were dealt with to study its population fluctuations in the Hokuriku district. Crude data were modified for distinguishing years of the low intensity of infestation by the insect from the rest of years. It is then clearly demonstrated that the low intensity of infestation were ordinarily preceded by heavy snowfall, although heavy snowfall could not be regarded as the only factor checking the vigorous multiplication of the populations.     

Mariculture of kurosoi,Sebastes schlegeli

Synopsis The technology of collecting developing larvae from female kurosoiSebastes schlegeli, and raising the larvae to juveniles (100 mm total length (TL)) to be released into the oopen sea, is presented. Gravid females 40–46 cm TL were captured in May–June 1977–1980 and held in the laboratory until parturition. Fecundity of fish in this size range was 100 000–184 000. Larvae were sequentially fed rotifers,Artemia nauplii, and young sand lance,Ammodytes personatus, until reaching 25 mm; this required 35 days and yielded a survival rate of 50%. Thereafter, the fish were reared in separate size groups to avoid cannibalism. Minced or chopped sand lance and commercial food were provided until the final size of 100 mm was attained. The growth of juvenile kurosoi from 25 to 100 mm required 85 days, with a survival rate of 90%. The effect of released cultured fish on the local stock is being determined from information on the recapture of tagged fish.     

A Method of Determination of Penta-, Tetra-, Tri-, and Disaccharides from Xylan by Ion-exchange Chromatography

Bacillus subtilis B7, a tmrA mutant, shows both tunicamycin resistance and a-amylase hyperproductivity. The tmrA characters can be transferred simultaneously to recipient cells by DNA-mediated transformation. We found a typical gene amplification phenomenon in the tmrA transformants and B7 strain. The amplified unit, 16.3kb in size, covers a-amylase structural gene amyE to another tunicamycin resistance gene tmrB, which is located 9kb downstream of the amyE gene. About 10 repeating units are supposed to be tandemly repeated in the transformants. Amplification of the wild amyE and tmrB genes could be the cause of the α-amylase hyperproductivity and tunicamycin resistance of the tmrA transformants and B7 strain.     

Phylogenetic study of the Characinae (Teleostei: Characiformes: Characidae)

The Characinae is a subunit of the Characidae of special significance in including Charax, the type genus of the family and the order Characiformes. Twelve genera and 79 species have been traditionally assigned to the Characinae, but the subfamily still lacks a phylogenetic diagnosis. Herein, a data matrix including 150 morphological characters and 64 taxa (35 species representing all genera of the Characinae and 29 included in other lineages within the Characiformes) was submitted to two cladistic analyses that differ in the inclusion/exclusion of Priocharax due to the difficulty of coding most of the character states in the miniature species of this genus. Both analyses resulted in a non‐monophyletic Characinae and this subfamily is herein restricted to only seven of the original 12 genera forming the clade (Phenacogaster((Charax Roeboides)(Acanthocharax(Cynopotamus(Acestrocephalus Galeocharax))))), which is supported by ten non‐ambiguous synapomorphies and is more closely related to other genera of the Characidae than those traditionally placed in the subfamily. A second clade includes the members of the tribe Heterocharacini (Lonchogenys(Heterocharax Hoplocharax)) as the sister‐group of Gnathocharax, supported by seven non‐ambiguous synapomorphies. This clade is more closely related to a taxon formed by Roestes and Gilbertolus based on seven non‐ambiguous synapomorphies. Results do not corroborate a close relationship between Roestes–Gilbertolus and the Cynodontinae. Inclusion of the genus Priocharax suggests that it is related more closely to the Heterocharacini, but the profound modifications in its anatomy possibly related to ontogenetic truncations obscure a better understanding of its relationships. A new classification of the Characinae and the Heterocharacinae is proposed. © 2012 The Linnean Society of London, Zoological Journal of the Linnean Society, 2012, 165 , 809–915.     

Molecular cloning and characterization of L-galactose-1-phosphate phosphatase from tobacco (Nicotiana tabacum)

L-Galactose-1-phosphate phosphatase (GPPase) is an enzyme involved in ascorbate biosynthesis in higher plants. We isolated a cDNA encoding GPPase from tobacco, and named it NtGPPase. The putative amino acid sequence of NtGPPase contained inositol monophosphatase motifs and metal binding sites. Recombinant NtGPPase hydrolyzed not only L-galactose-1-phosphate, but also myo-inositol-1-phosphate. The optimum pH for the GPPase activity of NtGPPase was 7.5. Its enzyme activity required Mg2+, and was inhibited by Li+ and Ca2+. Its fluorescence, fused with green fluorescence protein in onion cells and protoplasts of tobacco BY-2 cells, was observed in both the cytosol and nucleus. The expression of NtGPPase mRNA and protein was clearly correlated with L-ascorbic acid (AsA) contents of BY-2 cells during culture. The AsA contents of NtGPPase over expression lines were higher than those of empty lines at 13 d after subculture. This suggests that NtGPPase contributes slightly to AsA biosynthesis.     

Ionizing radiation suppresses FAP-1 mRNA level in A549 cells via p53 activation

Ionizing radiation (IR) is known to upregulate cell surface Fas through p53 activation in various cells. However, the signaling pathway intermediating between p53 activation and cell surface Fas upregulation remains to be elucidated. Recently, Fas-associated phosphatase-1 (FAP-1) has been reported to associate with Fas and inhibit cell surface Fas expression. We evaluated the expression of FAP-1 mRNA following IR in A549 cells. Ionizing radiation inhibited the expression of FAP-1 mRNA. Pretreatment with p53 inhibitor pifithrin alpha cancelled the IR-induced downregulation of FAP-1 mRNA. These results suggest that IR-induced p53 activation may upregulate cell surface Fas via the down-modulation of FAP-1.