Demonstration of phenolic compounds in plant tissues by an osmium-iodide postfixation procedure

A simple procedure to stain phenols in plant tissues is described. Postfixation with an aqueous solution prepared by mixing 2 cc of 2% osmium tetroxide and 8 cc of 3% potassium iodide yields brilliant visualization of phenol-containing vacuoles in different tissues of plants (e.g., coffee, oak, tobacco and spruce) bearing high concentration of phenolic compounds. Areas bearing phenols become dark gray to black. Chemical experiments demonstrate that osmium-potassium iodide (Os-KI) mixture reacts rapidly with several naturally occurring plant phenols, developing black solutions from which black solids precipitate. Phenols containing omicron-dihydroxy groups react with Os-KI solution more rapidly than other structurally different phenols. Therefore, omicron-dihydroxy units in an aromatic ring seem to function as primary sites of reactivity with the osmium-iodide complexes.     

Demonstration of Phenolic Compounds in Plant Tissues by An Osmium-Iodide Postfixation Procedure

A simple procedure to stain phenols in plant tissues is described, Postfixation with an aqueous solution prepared by mixing 2 cc of 2% osmium tetroxide and 8 cc of 3% potassium iodide yields brilliant visualization of phenol-containing vacuoles in different tissues of plants (e.g. coffee, oak, tobacco and spruce) bearing high concentration of phenolic compounds. Areas bearing phenols become dark gray to black. Chemical experiments demonstrate that osmium-potassium iodide (Os-KI) mixture reacts rapidly with several naturally occurring plant phenols, developing black solutions from which black solids precipitate. Phenols containing o-dihydroxy groups react with Os-KI solution more rapidly than other structurally different phenols. Therefore, o-dihydroxy units in an aromatic ring seem to function as primary sites of reactivity with the osmium-iodide complexes.     

Consistent production of phenolic compounds byPenicillium brevicompactum for chemotaxonomic characterization

A consistently produced group of fungal secondary metabolites fromPenicillium brevicompactum has been purified and identified as the Raistrick phenols. These compounds are shown to exist separately as an equilibrium mixture in aqueous solutions. The Raistrick phenols have all been included in the metabolite profile ofP. brevicompactum. By means of thin layer chromatography-scanning and high performance liquid chromatography-UV diode array detection, the chromatographic and spectroscopic data can be used in the chemotaxonomic characterization of the fungus.     

杉木林生态系统中土壤酚类物质含量的变化规律

以天然更新常绿阔叶林、老龄杉木[ Cunninghamia lanceolata (Lamb.) Hook.]林(代表一代杉木林)、二代杉木萌芽天然更新林和二代杉木人工林作为杉木林生态系统转换模式的系列样地,研究了不同季节(3月、6月、9月和12月)4个样地0～20和20 ～ 40 cm土层土壤中酚类物质(包括总酚、复合态酚和水溶性酚)含量的变化规律.结果表明:各样地土壤总酚含量最高(278.40 ～3 012.98 μg·g-1),复合态酚含量次之(20.67～ 430.54 μg·g-1),水溶性酚含量最低(0.36～6.01 μg·g-1).各样地问不同土层中总酚、复合态酚和水溶性酚含量均值的高低变化不一致,但总体上顺着森林生态系统转换的方向(天然常绿阔叶林→一代杉木人工林→二代杉木人工林),0～20和20～ 40 cm土层中总酚和复合态酚含量以及O～20 cm土层中水溶性酚含量均值均呈现逐渐增加的趋势,而20～40 cm土层中水溶性酚含量均值则无明显变化规律.各样地0～20 cm土层中各种酚类物质含量总体上高于20～40 cm土层,表明随土层加深,总酚、复合态酚和水溶性酚含量呈现逐渐降低的趋势.各种酚类物质含量具有明显的季节变化规律,总酚和复合态酚含量总体上呈现出3月和9月较高、6月和12月较低的变化趋势,水溶性酚含量总体上呈现出随着季节变化(3月至12月)逐渐降低的趋势,不同季节间土壤中各类酚类物质含量的差异均达到了极显著水平.研究结果显示:随杉木人工林栽植代数的增加,土壤中会积累一定量酚类物质,但其中水溶性酚含量远低于使植物中毒的水平(50 μg·g-1),造成杉木中毒的可能性较小.     

Recent problems and developments in the control of post-harvest deterioration of softwood timber by chemical treatments

In the last ten years there has been a slow movement away from the use of chlorinated phenols for the prevention of post-harvest deterioration of timber. Experience has shown that several hazard situations exist which together with other factors affect the activity of chemicals used to replace the chlorinated phenols. No one chemical is likely to dominate the market again and much scope exists for improving the performance of biocides in this application.     

Apparent stimulation of dibenzo[a,e]fluoranthene in vitro metabolism in the presence of norharman

The effects of norharman (NH) on the metabolism of dibenzo[a,e]-fluoranthene (DBF) and on its fixation on DNA, RNA and proteins have been studied in vitro by incubation with S-9 and microsomes from rats and mice. NH causes a decrease of the activity of microsome monooxygenases proportionally to its concentration but has no effect on the activity of NADPH P-450 reductase nor on that of epoxide hydrolase. Paradoxically, the amount of DBF hydrophobic metabolites and especially that of diols and phenols, increases in the incubation mixture in the presence of NH; this increase is independent of the presence of conjugation enzymes of cytosol. NH does not modify the covalent binding of DBF on the microsome RNA, conversely it decreases the binding of DBF on the DNA and on the proteins of the incubation mixture. This could partly explain the increase of DBF diols and phenols by an accumulative effect. Two higher homologs of NH: benzo[g]-beta carboline and benzo[i]-beta carboline, tested under the same conditions, proved inhibitory.     

The content of phenolic compounds in medicinal plants of a botanical garden (Kaliningrad oblast)

The total content of phenolic compounds and antioxidants has been determined in medicinal plants (66 species from 31 families). Promising plant species with high antioxidant activity and maximum content of phenols have been found. Based on these data, the plants can be used as a basis for making innovative functional food products with an increased antioxidant effect.     

Isolation and Identification of the Phenols of Paul's Scarlet Rose Stems and Stem-Derived Suspension Cultures

The phenols of Paul's Scarlet rose stems and stem-derived cell cultures have been analyzed using C₁₈-reversed-phase high performance liquid chromatography.

Rose stems were found to contain gallic acid, (+)catechin, (−)epicatechin, the dimers (−)epicatechin-(+)catechin and (+)catechin-(+)catechin, a polymeric procyanidin, ferulic acid, and several gallotannins. In contrast, a cell suspension of Paul's Scarlet rose which has been maintained in culture for over 25 years contained only low levels of gallic acid and (−)epicatechin-(+)catechin. The phenol content of a second rose cell line which was started from the same initial isolate in 1957, but which was maintained in a laboratory other than our own was quantitatively and qualitatively similar to the cell line kept in our laboratory for the last 20 years. A third cell line which we started 6 months ago contained a wide variety of phenols, most of which were in common with those of rose stems.

Selective subculturing of smaller cell clumps of our oldest cell line failed to enhance either the quantities or the diversity of phenols which accumulated in these cultured cells. Possible reasons for the failure of selective subculturing to enhance phenol levels in this long-established cell line are discussed.

     

Effect of phenols on the oxidation of estradiol by uterine peroxidase.

The effect of various phenols on the conversion of [4 -14C]estradiol to water-soluble products by estrogen-induced uterine peroxidase (EC 1.11.1.7) has been investigated. Evidence was provided that those phenols which enhanced the oxidation of estradiol exerted their effect by activating peroxidase or protecting the enzyme from inactivation by the products of the reaction rather than by inhibiting the breakdown of hydrogen peroxide by catalase (EC 1.11.1.6). It has also been shown that tyrosine acted both as an activator of uterine peroxidase and as a water-soluble acceptor for the metabolites of estradiol. The ability of tyrosyl peptides to form conjugates with estradiol was influenced by the other amino acids and decreased with the number of adjacent tyrosyl residues.     

Vaseline-peptone mixture as a stimulant of ascites formation in mouse hybridoma carriers

A preliminary injection of vaseline oil with 3% of peptone in a dose of 0.5 ml induced ascitic tumors with a high taking rate in mice used as recipients of hybridomas. The dependence of the amount of ascitic fluid produced in such animals and the content of monoclonal antibodies on the time of the preliminary injection of the above-mentioned mixture, the number of cells introduced into the recipient animal, and the time of ascites collection has been studied. The optimal scheme of obtaining ascites with the use of the proposed mixture is presented. The comparison of the mixture with other stimulants of ascites production has been made.     

Influence of the composition of essential oils on their antioxidant and antiradical properties

The antiradical and antioxidant properties of essential oil mixtures (EOMs) with various compositions were studied and compared in model systems of hexanal autooxidation, thermal oxidation of methyl linoleate and beta3-carotene, and in a reaction with the stable diphenylpicrylhydrazyl radical. It was found that all EOMs under investigation exhibited antiradical activity. The highest antiradical activity was observed for an EOM containing monoterpene hydrocarbons as the main components (the phenol content was low). The antioxidant activity of all EOMs was from 60 to 98% and depended on the model system composition and the method of assay. The mixture with the highest phenol content exhibits the highest antioxidant activity level in the hexanal autooxidation system. EOMs with a high content of phenols and alpha- and y-terpinenes were the most efficient antioxidants in the beta3-carotene model system. The study confirmed the possibility to vary the antioxidant and antiradical properties of essential oils by preparing their mixtures with a specific composition.     

Exogenous phenol increase resistance of Ulmus minor to Dutch elm disease through formation of suberin-like compounds on xylem tissues

The survival of some elms to Dutch elm disease (DED) epidemics could be related with the application of disinfectant products based on simple phenols. To test this hypothesis, the protective effect of different phenolic treatments in Ulmus minor trees was evaluated through inoculations with Ophiostoma novo-ulmi, the current DED pathogen. During spring 2004 and spring 2005, 4-year-old elms were: (i) watered with a 0.02% solution of the phenolic fraction of phenolic oil, (ii) watered with a 0.02 and 0.2% solution of a phenol–cresol mixture, and (iii) trunk injected with a 0.2% solution of phenol–cresol mixture. In May, trees were artificially inoculated with O. novo-ulmi. At the end of the 2004 and 2005 vegetative periods, phenol-treated trees showed significantly lower wilting values than control trees. One week of bud break delay was observed in trees watered with the 0.2% solution of phenol–cresol mixture. Fourier transform-infrared spectroscopy and electrospray ionisation mass spectrometry evidenced enhanced levels of suberin-like compounds in phenol-treated trees with respect to non-treated trees. The deposition of suberin in xylem tissues, as a response to phenol treatments, might be considered as one of the mechanisms of resistance of elms to O. novo-ulmi.     

Enzymatic modification of kraft lignin through oxidative coupling with water-soluble phenols

The aromatic polymer lignin can be modified through promotion of oxidative coupling between phenolic groups on lignin and various phenols. The reaction is initiated by an oxidation of both components, e.g., by using the oxidoreductases laccase or peroxidase. Coupling between phenolic monomers and lignin has previously been studied by the use of radio-labeled phenols. In this study, incorporation of water-soluble phenols into kraft lignin, using laccase as catalyst, was investigated. Several phenols with carboxylic or sulfonic acid groups were used as markers for the incorporation. The modified lignin was isolated and the amount of phenol incorporated was characterized by means of titration, quantitative 1H-NMR, and quantitative 31P-NMR after modification with 2-chloro-4,4,5,5-tetramethyl-1,2,3-dioxaphospholane. Only a few of the phenols studied were found to be incorporated into lignin. When the phenol guaiacol sulfonate was incorporated into kraft lignin, the lignin became water-soluble at pH 2.4 and a low ionic strength due to the introduction of sulfonic acid groups. The content of sulfonic acid groups in the product was 0.5-0.6 mmol/g lignin. A lower amount of 4-hydroxyphenylacetic acid was incorporated under similar conditions.     

Forest Gaps Alter the Total Phenol Dynamics in Decomposing Litter in an Alpine Fir Forest

The total phenol content in decomposing litter not only acts as a crucial litter quality indicator, but is also closely related to litter humification due to its tight absorption to clay particles. However, limited attention has been focused on the total phenol dynamics in foliar litter in relation to forest gaps. Here, the foliar litter of six representative tree species was incubated on the forest floor from the gap center to the closed canopy of an alpine Minjiang fir (Abies faxoniana) forest in the upper reaches of the Yangtze River and eastern Tibetan Plateau. The dynamics of total phenol concentration in the incubated litter was measured from November 2012 to October 2014. Over two-year incubation, 78.22% to 94.06% of total phenols were lost from the foliar litter, but 52.08% to 86.41% of this occurred in the first year. Forest gaps accelerated the loss of total phenols in the foliar litter in the winter, although they inhibited the loss of total phenols during the growing season in the first year. In comparison with the effects of forest gaps, the variations of litter quality among different species were much stronger on the dynamics of total phenols in the second year. Overall, the loss of total phenols in the foliar litter was slightly higher in both the canopy gap and the expanded gap than in the gap center and under the closed canopy. The results suggest that the predicted decline in snow cover resulting from winter warming or vanishing gaps caused by forest regeneration will retard the loss of total phenol content in the foliar litter of alpine forest ecosystems, especially in the first decomposition year.     

The theoretical basis and practical application of a new method of selection for determining water deficiency in plants

Summary 1. On the basis of new theoretical considerations developed by us a new and reliable method has been elaborated for the rapid determination of water deficit in plant cultures. 2. In paper-chromatograms of plants optimally supplied with water free proline is shown to be present in very small amounts, usually only as traces. Comparative experiments showed that in cases of water shortage the proline content of the leaves of the plant strikingly increases. This commonly occurs in every phase of development of mono- and dicotyledons. 3. Drought-resistant plants synthetize more proline than varieties less resistant to water deficit when equally water deficient. 4. By a method which does not require chromatography the procedure of tests has been accelerated so that leaf extract containing amino acid mixture was dropped onto a chromatogram paper and immediately treated with isatine. In this test the effect of slight, permanent or medium water deficit upon proline content is so great that extracts react with isatine to give an intense dark blue colour which suppresses the reddish-brown colour of other amino acids. 5. To avoid the necessity of preparing isatine reagent frequently, because it loses its amino acid sensitivity very rapidly, a new isatine solution has been formulated. Our isatine-paper indicator prepared with this solution keeps its reactivity with amino acids 5 to 6 months. 6. An unusually high proline concentration occurs also if the salt content of irrigation water or also of the soil is high. The increase of the proline level indicates therefore physiological dryness.     

Bacteriostatic activity of some coumarin derivatives

The bacteriostatic effect of some mono- and dihydroxy-coumarins and isopropyldihydrofuranocoumarins have been examined and compared with other simple phenols. Most of the coumarins proved to be far more bacteriostatic than the simple phenols.     

Formation of Fine Oil-in-Water Emulsions by the Gel Emulsification Method with Saccharide and Protein

An emulsification method using a gel-like phase of a saccharide and protein mixture has been developed. In the method, which is called a gel emulsification method, an oil is added to the highly concentrated saccharide solution containing protein to form a clear gel-like phase, which followed by dilution with water to form a fine oil-in-water emulsion. This emulsion was investigated as to its emulsifying activity and emulsion stability as compared with that obtained by high-shear equipment, which was called a homomixer method. The emulsifying activity of the emulsions prepared by the gel emulsification method was much higher than that of the emulsions prepared by the homomixer method.

The emulsions prepared by both methods were highly stable in terms of the stability against coalescence. On the other hand, the stability against creaming of the emulsions prepared by the gel emulsification method was much higher than that of the emulsions prepared by the homomixer method.

The surface hydrophobicity of the protein and the unfreezable water content in the highly concentrated saccharide solution containing protein were not correlated to the emulsifying properties of the emulsions prepared by the gel emulsification method, which appeared to be dependent on the viscosity of the highly concentrated saccharide solution containing protein.     

Influence of the composition of essential oils on their antioxidant and antiradical properties

The antiradical and antioxidant properties of essential oil mixtures (EOMs) with various compositions in model systems of hexanal autooxidation, thermal oxidation of methyl linoleate and β-carotene, and in a reaction with the stable diphenylpicrylhydrazyl radical were studied and compared. It was found that all studied EOMs exhibited antiradical activity. The highest antiradical activity was observed for an EOM containing monoterpene hydrocarbons as the main components (the phenol content was low). The antioxidant activity of all EOMs was from 60 to 98% and depended on the model system composition and the method of assay. The mixture with the maximum phenol content exhibits the highest antioxidant activity level in the hexanal autooxidation system. EOMs with a high content of phenols and α- and γ-terpinenes were the most efficient antioxidants in the β-carotene model system. The study confirmed the possibility to vary the antioxidant and antiradical properties of essential oils by preparing their mixtures with a specific composition.     

The Role of Tannins in Neutral Red Staining of Pine Leaf Vacuoles

The vital stain, neutral red, was retained by the cell vacuoles of white pine, Pinus strobus L., through the fixing and embedding process. The results obtained were equivalent to living tissue when the material was fixed in 10% formalin or Bouin's solution and embedded in wax. Chromatographic studies indicated that water-soluble tannins were responsible for adsorption of the neutral red, but only after treatment with formaldehyde. Analyses with other mono-, di-, and trihydroxy phenols suggested that ferric ions and phenols with at least 3 hydroxyl groups per molecule were essential to the reaction.