Formation of Hydrogen Peroxide by NAD(P)H Oxidation with Isolated Cell Wall-Associated Peroxidase from Cultured Liverwort Cells, Marchantia polymorpha L.

Hydrogen peroxide was formed in isolated cell walls from Marchantiapolymorpha L. in the presence of MnCl₂ by either NADH or NADPHoxidation. This reaction was stimulated by phenols such as 2,4-dichlorophenolor p-coumarate, suggesting a reaction similar to that proposedfor the last step of lignification in higher plant cells, althoughbryophytes have been reported to be devoid of lignin. (Received June 16, 1987; Accepted March 3, 1987)     

Wavelength Effect on the Action of a N-Phenylimide S-23142 and a Diphenylether Acifluorfen-Ethyl in Cotyledons of Cucumber (Cucumis sativus L.) Seedlings

Specific wavelengths of light required for expression of phytotoxic activity of S-23142 (N-[4-chloro-2-fluoro-5-propargyloxy]phenyl-3,4,5,6-tetra- hydrophthalimide) and acifluorfen-ethyl (ethyl-5-[2-chloro-4-(trifluoromethyl)phenoxy]-2-nitro benzoic acid) were determined in cotyledons of cucumber seedlings using the Okazaki Large Spectrograph. Leakage of amino acids from the cotyledons was measured as an indication of the phytotoxic activity. The wavelength effects showed common major peaks of activity at 550 and 650 nanometers and a minor peak at 450 nanometers for both herbicides, indicating a common primary photoreaction. Concomitant application of DCMU (3-[3,4-dichlorophenyl]-1,1-dimethylurea) with S-23142 had little influence on the effective wavelengths for S-23142 activity. Light of 450 and 650 nanometers was relatively less effective in achlorophyllous tissue grown in far red light than in green tissue. These results strongly suggest that the phytotoxic action of S-23142 and diphenylethers involves multiple photoreactions and that one of the photoreceptor pigments may be chlorophyll or its related pigment, although photosynthesis is not involved.     

Regulation of retrochalcone biosynthesis: Activity changes of O-methyltransferases in the yeast extract-induced Glycyrrhiza echinata cells

Three O-methyltransferases which catalyze S-adenosyl-L-methionine (SAM)-dependent O-methylation of licodione (LMT), flavone/flavonol (FMT), and caffeic acid (CMT) were separated from the callus culture of Glycyrrhiza echinata, and characteristic differences between their pH optima and Mg²⁺ requirement for activity were demonstrated. The activity of LMT, which is involved in retrochalcone (echinatin) biosynthesis, but not of FMT or CMT, was found to be stimulated when suspension-cultured G. echinata cells were treated with yeast extract (YE), which causes rapid production of echinatin in the cells. Cycloheximide suppressed both the YE-induced echinatin formation and LMT enhancement. The results indicate a selective induction of retrochalcone pathway in Glycyrrhiza cells in response to stress.Abbreviations SAM S-adenosyl-L-methionine - LMT, SAM licodione 2-O-methyltransferase - FMT, SAM flavone/flavonol O-methyltransferase - CMT, SAM caffeate 3-O-methyltransferase - OMT O-methyltransferase - CH cycloheximide - YE yeast extract This paper is Part 47 in the series Studies on Plant Tissue Cultures. For Part 46, see Ayabe S, Iida K, Furuya T (1986) Phytochemistry: in press     

Linear alpha-human atrial natriuretic peptide analogs display receptor binding activity and inhibit alpha-hANP-induced cGMP accumulation

We have synthesized a series of [Cys(R)7,23]alpha-hANP analogs, in which the two Cys residues were modified with various alkyl groups(R); i.e., R=Acm, Pe, Qe, Cam, Me, Ae, Bzl, Cm, Ocam and sulfo. The Acm-, Cam-, and Me-analogs exhibited binding activity as potent as alpha-hANP in rat vascular smooth muscle cells (VSMC). Binding activity of the analogs decreased progressively as the bulkiness of the R group increased. None of the analogs caused accumulation of cGMP in VSMC and vasorelaxant activity in rat aorta. Acm-, Cam- and Me-analogs substantially antagonized alpha-hANP-induced cGMP accumulation, but did not antagonize vasorelaxation induced by alpha-hANP in vitro.     

The primary structure of human EGF produced by genetic engineering, studied by high-performance tandem mass spectrometry

The primary structure of human epidermal growth factor (hEGF), which was produced by Escherichia coli using recombinant DNA technique, has been studied by tandem mass spectrometry. The molecular weight of hEGF (about 6200 amu) was determined by fast atom bombardment mass spectrometry. Then reduced and carboxymethylated hEGF was digested by chymotrypsin into seven peptides which could cover the whole sequence of hEGF. The amino acid sequences of five of these seven peptides could be confirmed by tandem mass spectrometry with or without isolation by high-performance liquid chromatography (HPLC). After isolation by HPLC, the other two peptides were digested with trypsin or thermolysin into small peptides, and sequenced by tandem mass spectrometry.     

Primary photoprocesses of phytochrome. Picosecond fluorescence kinetics of oat and pea phytochromes

The primary photoprocesses of etiolated oat and pea phytochromes (Pr forms) are diffusion-modulated by the microscopic viscosity within the chromophore pocket. The chromophore pocket is preferentially accessible to glycerol but not to Ficoll. Glycerol preferentially retarded the rate (rate constant ca. 1-2 X 10(10) s-1) of the initial reaction from the Qy excited state of phytochrome, whereas it increased the long fluorescence lifetime (nanosecond) component that can be attributed to either an emitting intermediate or to modified/conformationally heterogeneous phytochrome populations. The picosecond time-resolved fluorescence spectra of different phytochrome preparations (i.e., full-length vs 6/10-kDa NH2-terminus truncated forms of phytochromes from monocot and dicot plants) revealed no significant differences. The spectra in the picosecond time scale showed no spectral shifts, but at longer time scales of up to approximately 1.90 ns, significant blue spectral shifts were observed. The shifts were more in the truncated than in the full-length pea phytochrome. Comparison of the fluorescence decay data and the picosecond time-resolved fluorescence spectra suggests differences in conformational flexibility/heterogeneity among the preparations of the monocot vs dicot phytochromes and the full-length native vs the amino terminus truncated phytochromes.     

Regional distribution of DNA and RNA in rat brain: A sensitive determination using high-performance liquid chromatography with electrochemical detection

DNA and RNA contents in 20 brain regions or nuclei of the rat were determined by a highly sensitive method using high-performance liquid chromatography with electrochemical detection. The high DNA and RNA contents were found in the hypothalamic nuclei, especially the median eminence-arcuate nucleus. These results may be available for the preparation of nucleic acids as the regional control.     

Production of theanine and other γ-glutamyl derivatives by Camellia sinensis cultured cells

The maximum theanine production by Camellia sinensis cultured cells was achieved by culturing in the modified MS medium containing 2 mg/l indole-3-butyric acid, 0.1 mg/l kinetin, 0 mM NH₄NO₃ and 39.6 mM KNO₃ with 40 mM ethylamine hydrochloride or 20 mM ethylamine hydrochloride and 10 mM L-glutamic acid. Other primary amines, such as methylamine, n-butylamine, 2-hydroxyethylamine, 2cyanoethylamine, aniline, benzylamine and phenylethylamine, were also biotransformed to N⁵-alkyl-L-glutamine derivatives by C. sinensis cultured cells.Abbreviations IBA indole-3-butyric acid - K kinetin - MS medium Murashige and Skoog's basal medium (1962) - NMR nuclear magnetic resonance Part 70 in the series Studies on Plant Tissue Culture. For Part 69 see Furuya et al. (1990).     

Analysis of restriction profiles of Mitochondrial DNA from Sporothrix schenckii and related fungi

Restriction profiles by HaeIII of mitochondrial DNA were studied for classification and distinction of Sporothrix schenckii (100 strains), S. schenckii var. luriei (1), S. curviconia (1), S. inflata (7), Ceratocystis stenoceras (17) and C. minor (7). These 6 species showed unique restriction profiles which could be discriminated from each other. S. schenckii was further separable into 11 types, S. inflata into 4 types, C. stenoceras into 4 types and C. minor into 7 types based on restriction profile heterogeneity.