Assimilation of berberine and chelidoxanthine by bacteria

Summary Several species of bacteria are able to assimilate the alkaloid berberine while growing in extracts ofBerberis vulgaris, Chelidonium majus, andMahonia aquifolium. They can also assimilate the alkaloid in nutrient media containing berberine hydrochloride in concentrations up to and including 2.0 per cent, but only if provided with suitable sources of carbon and nitrogen. Some bacteria, especially Gram-positive species, were found to be sensitive to relatively low concentrations of the alkaloid. The alkaloid appeared to be assimilated during the early stages of growth, and to be held in the protoplasm, for surface growth on agars and the washed cells had the bright yellow fluorescence of berberine, in ultra violet light. In mineral salts media with glucose and with amino acids as sources of nitrogen the intake of the alkaloid appeared to be stimulated by aspartic acid, glutamic acid, cysteine, methionine and taurine. Bacterial cells washed from colonies on agar containing the alkaloid served as a source of nitrogen to other bacteria, which extracted and assimilated the berberine from the washed cells.Contribution from the Faculty of Agriculture, Macdonald College, Que., Canada, Journal Series No. 403.The work formed part of that done by the junior author in partial fulfillment of the requirements for the Ph. D. degree of McGill University.     

A berberine-aniline blue fluorescent staining procedure for suberin,lignin, and callose in plant tissue

Summary A fluorescent staining procedure to detect suberin, lignin and callose in plants has been developed. This procedure greatly improves on previous methods for visualizing Casparian bands in root exodermal and endodermal cells, and performs equally well on a variety of other plant tissues. Berberine was selected as the most suitable replacement forChelidonium majus root extract after comparing the staining properties of the extract with those of four of its constituent alkaloids. Aniline blue counterstaining efficiently quenched unwanted background fluorescence and nonspecific berberine staining, while providing a fluorochrome for callose. When used with multichambered holders which allow simultaneous processing of freehand sections, this efficient staining procedure facilitates morphological studies involving large numbers of samples.Abbreviations ISCC-NBS Inter-Society Color Council-National Bureau of Standards - UV ultraviolet light     

The inhibition enzymatic hydrolysis of acetylthiocholine by acetylcholinesterase using principal alkaloids isolated from celandine and macleya and their derivatives

A study was made of a possible inhibitory action on the enzymatic hydrolysis of acetylthiocholine by human erythrocyte acetylcholinesterase of principal alkaloids isolated from Chelidonium majus L. and Macleaya (Bocconia) cordata and microcarpa (namely sanguinarine, chelidonine, berberine), and of drugs "Ukrain" (thiophosphoric acid derivative of a sum of the alkaloids isolated from Chelidonium majus L.) and "Sanguirythrine" (a mixture of unseparated closely related to benzo[c]phenanthridine alkaloids sanguinarine and chelerythrine, isolated from Chelidonium majus L. and other plants of Papaveraceae family). All agents under study have been shown to be reversible inhibitors of the enzymatic hydrolysis of acetylthiocholine. On the basis of the kinetic data it has been determined that chelidonine belonged to reversible inhibitors of a competitive type. All other examined agents have been demonstrated to be inhibitors of a mixed competitive-noncompetitive type, and a greater contribution to the inhibition was made by the competitive constituent. Among all examined agents berberine, sanguinarine and "Sanguirythrine" were the strongest inhibitors of this reaction (the values of generalized inhibitory constants being 0.23, 0.23 and 0.29 microM, respectively) and cheliodonine and "Ukrain" were much weaker (2.0 and 2.5 microM, respectively). Judging from the data obtained, sanguinarine and chelerythrine exert similar inhibitory effects on the reaction of enzymatic hydrolysis of acetylthiocholine, since sanguinarine and "Sanguirythrine" have nearly equal generalized inhibitory constants.     

Life cycle and morphology ofMelampsora yezoensis onSalix serissaefolia

Uredinial and telial states of aMelampsora species occurring on the leaves ofSalix serissaefolia were for the first time recorded. Field observations and inoculation experiments showed that the spermogonial and aecial states of the fungus were formed onChelidonium majus var.asiaticum andCorydalis incisa. The fungus was identified asMelampsora yezoensis based on the morphological observation of all the spore states. Urediniospores were able to infectS. erocarpa andS. pierotii as well asS. serissaefolia andS. jessoensis. Salix serissaefolia, S. eriocarpa, andS. pierotii are new uredinial and telial hosts ofM. yezoensis, andC. majus var.asiaticum andC. incisa are new spermogonial and aecial hosts of the fungus. Contribution No. 124, Laboratories of Plant Pathology and Mycology, Institute of Agriculture and Forestry University of Tsukuba.     

Seed desiccation limits removal by ants

Ants collect and disperse seeds that bear an attractive nutritive body called the elaiosome. In mesic habitats, many myrmecochorous plant species have elaiosomes that are usually soft and desiccation-sensitive. The aim of this study was to link the desiccation rate of two species of seeds (Chelidonium majus and Viola odorata) to the seed-removing behaviour of the ant Myrmica rubra. In laboratory experiments seeds of both species lost one-third of their weight in 24 h. Concurrently, seed removal rates decreased sharply (92%) for Viola odorata over 24 h and slowly for Chelidonium majus, which retained one-third of its attractiveness after one month of desiccation. Seeds recovered their initial weight almost entirely after being soaked in water for 18 h. This rehydration partially (Viola odorata) or totally (Chelidonium majus) restored the attractiveness of the seeds. In this paper, we show that the window of seed attractiveness during which ant–plant interactions occur is desiccation-dependent and plant-specific.     

Effect of some isoquinoline alkaloids on enzymatic activity of acetylcholinesterase and monoamine oxidase

It has been shown, that some benzo[c]-phenanthridine and diisoquinoline alkaloids isolated from Chelidonium majus L. and Macleaya (Bocconia) cordata and M. microcarpa (berberine, sanguinarine, chelidonine) and of drugs ("Ukrain" and "Sanguirythrine") inhibited the enzyme activity of acetylcholinesterase from human erythrocyte and monoamine oxidase from the rat liver. All agents under study have been shown to be reversible inhibitors of the enzymatic hydrolysis of acetylthiocholine. It has been determined that chelidonine belonged to reversible inhibitors of a competitive type, all other examined agents have been demonstrated to be inhibitors of a mixed competitive-noncompetitive type, and a greater contribution to the inhibition was made by the competitive constituent. Among all examined agents berberine, sanguinarine and "Sanguirythrine" were the strongest inhibitors of this reaction and chelidonine and "Ukrain" were much weaker. All agents under study have been shown to be irreversible inhibitors of the oxidative deamination reaction of serotonine and tyramine and not to influence the oxidative deamination reaction of benzylamine as a substrate. Among the examined agents, alkaloid sanguinarine and drug "Ukrain" are the strongest inhibitors of the reaction, alkaloids berberine, sanguinarine and "Sanguirythrine" exhibit a weaker action.     

Correlation of the cytotoxic activity of four different alkaloids, from Chelidonium majus (greater celandine), with their DNA intercalating properties and ability to induce breaks in the DNA of NK/Ly murine lymphoma cells

The purpose of this study was to examine the relationship between the DNA intercalating characteristics and the DNA damaging capacity of four alkaloids extracted from Chelidonium majus L, as well as their toxicity towards murine NK/Ly lymphoma cells. Chelerythrine, sanguinarine and coptisine were found to be intercalated into the DNA isolated from NK/Ly cells, meanwhile, chelidonine exhibited no affinity to DNA. Sanguinarine exhibited the greatest toxicity toward NK/Ly cells, and the toxicity of the other three decreased in descending order: chelerythrine, coptisine and chelidonine. Chelerythrine and sanguinarine caused DNA damage, illustrated by the formation of comets of the third class. Coptisine was less toxic than chelerythrine and sanguinarine, and affected the formation the same class of comets in higher concentration. The quantity of comets induced by chelidonine were negligible, a finding consistent with its inability to intercalate into DNA structure. The ability of four main alkaloids of Chelidonium majus L., to intercalate into DNA isolated from murine NK/Ly lymphoma cells, correlated with their ability to induce breaks in cellular DNA and with their toxic effect towards those cells.     

Phylogenetic relationships of Coreanomecon (Papaveraceae: Chelidonioideae) inferred from seed morphology and nrITS sequence data

The phylogenetic position of the Coreanomecon, a monotypic genus endemic to Korea, has been controversial for a long time. Coreanomecon has variously been placed in its own genus or combined with Hylomecon or Chelidonium. The main purpose of this study was to examine the phylogenetic position of Coreanomecon in relation to genera of the subfamily Chelidonioideae using seed morphology and molecular data. The seed morphology of 10 genera of Chelidonioideae was examined using microtome sections and scanning electron microscopy. The shape and size of exotestal cells varied between genera. The exotestal cells were tangentially elongated in Chelidonium majus, Stylophorum diphyllum and Hylomecon vernalis, whereas the cells were sub‐orbicular and broad in Coreanomecon hylomeconoides. The endotesta was represented by thick palisade‐like cells that contain small rectangular crystals near their outer walls. The mesotesta was totally collapsed in Co. hylomeconoides but wholly or partly persistent in Ch. majus and H. vernalis. The seed surface of Co. hylomeconoides was well ornamented with a unique echinate seed surface, whereas other genera showed plain and reticulate seed surfaces. A phylogenetic analysis of the genera including Coreanomecon was conducted with nuclear ribosomal internal transcribed spacer (nrITS) sequences using genetic distance, maximum parsimony, maximum likelihood and Bayesian inference methods. The results confirm that Co. hylomeconoides is separated from both Hylomecon species and is a sister group to Chelidonium majus and Stylophorum diphyllum with robust bootstrap support. In addition to differences in the absence of cauline leaves and rhizomes, the presence of a hairy stem and leaves, and 12‐pericolpate pollen, seed characters and molecular data strongly support the recognition of Coreanomecon as an independent genus, distinct from Hylomecon and Chelidonium.     

Induction of adventitious buds in cultured petal explants ofChelidonium majus L.

Petal explants ofChelidonium majus L. (Papaveraceae) formed noteworthy adventitious buds without any intermediate callus when cultured under appropriate conditions. Bud formation was favored by combinations of 1–2 mg/l indoleacetic acid (IAA) and/or 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.1–0.5 mg/l kinetin (K). In the present study, neither bud formation nor callus formation occurred in cultures of excised leaves. A histological study revealed that adventitious bud formation occurred only in single epidermal layers of petals, while several subepidermal parenchyma layers did not join in its formation. Activation zones arising from the epidermis underwent intense cell divisions to initiate buds on the epidermal surface. These buds later turned green in color, developing into shoots which eventually grew into plantlets after root formation.     

Correlation of Dispersibility of Proteins with that of Selenium in Teas

Selenium-enriched tea was suggested as a possible source of supplemental Se. The result of this study indicates that it is not practicable to make selenium-enriched tea as a beverage like traditional green tea or black tea for the supplementation of selenium in human diet. The selenium dispersibilities of fresh tea leaves, green tea, and black tea highly correlated with those of protein (r ² = 0.998). The high protein dispersibility (85.0%) of fresh tea leaves in water solution was accompanied by that of selenium (93.8%). Decreases in protein dispersibility of green tea and black tea to 2.5% and 4.2 % coincided with those of selenium to only 8.3% and 10.1%, respectively. The amount (14.90 μg) of selenium in saturated ammonium sulphate (a protein precipitating reagent) precipitate was 83.8% of that (17.79 μg) in fresh tea leaf extract, and after the saturated ammonium sulphate precipitate was dialyzed against distilled water overnight, the amount (14.37 μg) of selenium remaining in the dialyzed precipitate (protein) was still 80.8% of that in the fresh tea leaf extract. However, there were no significant differences (p > 0.05) between the amount of selenium in the saturated ammonium sulphate precipitate and that in the saturated ammonium sulphate precipitate that was dialyzed.     

盐酸黄连素对蚜虫生物活性的研究

黄连素为黄连Coptis chinensis Franch体内的主要生物碱成份。用采自贵阳花溪附近的新鲜黄连主根提取盐酸黄连素,将粗品重结晶一次后,用 HITACHI-220紫外可见分光光度计测得一次重结晶产品含盐酸黄连素91%,黄连主根中的黄连素含量为11.56%。室内生物测定结果表明,盐酸黄连素对甘蓝蚜Brevicoryne brassicae (L.)具有很强的触杀作用和较好的拒食活性。1%盐酸黄连素的乙醇溶液点滴蚜虫,96 h死亡率高达92.5%,校正死亡率为91.4%, LC₅₀值为(2.289±0.0125)g/L,回归方程Y=7.712+1.027X,相关系数r为0.958,达显著水平;1%盐酸黄连素的乙醇溶液处理叶片,24 h处理和对照叶面的蚜虫栖息率分别为20.6%和79.4%,拒食效果为74.04%, AFC₅₀值为(4.055±0.0215) g/L ,回归方程Y＝9.115+1.719X,相关系数r为0.998,达极显著水平。     

Inhibition of liver mitochondrial monoamine oxidase activity by alkaloids isolated from Chelidonium and Macleaya and by their derivative drugs

It has been shown that the major alkaloids from plants Chelidonium majus L. and Macleaya (Bocconia) cordata and microcarpa, namely, berberine, sanguinarine, chelidonine, and drugs "Ukrain" (thiophosphoric acid derivative of a sum of the alkaloids isolated from Ch. majus L.) and "Sanguirythrine" (a mixture of the alkaloids sanguinarine and chelerythrine, w/w 3:7, isolated from Macleaya), are irreversible inhibitors of oxidative deamination reaction of serotonin and tyramine as substrates, catalyzed by rat liver mitochondrial monoamine oxidase (MAO). At the same time these substances do not influence the oxidative deamination reaction of benzylamine as substrate (in concentration 1 mM or less). The substrate specificity of this inhibition manifests that mainly the oxidative deamination reactions catalyzed by MAO form A are inhibited by the agents studied. Among the examined agents, alkaloid chelidonine and drug "Ukrain" are the strongest inhibitors of the reaction. Alkaloids berberine and sanguinarine and drug "Sanguirythrine" exhibit a weaker action. Judging from the data obtained, sanguinarine and chelerythrine appear to exert similar inhibitory effects in this reaction, since sanguinarine and "Sanguirythrine" have similar values of bimolecular rate constants of their interaction with mitochondrial MAO. As it is well known, the MAO inhibitors appear to be, as a rule, pronounced antidepressants. The combination of malignotoxicity and antidepressive activity in drug "Ukrain" seems to be favourable for its clinical applications.     

Permeability of the suberized mestome sheath in winter rye

Mestome sheath cells of winter rye (Secale cereale L. cv Puma) deposit suberized lamellae in their secondary cell walls. Histochemical tests including acid digestion and staining with Sudan IV and Chelidonium majus root extract were used to detect the presence of suberin in the primary cell wall. There was no evidence of a Casparian band between adjacent mestome sheath cells. Fluorescent dye techniques were used to trace solute movement through the rye leaf apoplast. Calcofluor white M2R, a fluorescent dye which binds to cell walls as it moves apoplastically, proved to be too limited in its mobility in leaves to test mestome sheath permeability. Trisodium 3-hydroxy-5,8,10 pyrene trisulfonate, a fluorescent dye which is mobile in the apoplast, moved easily up the vascular bundles in the transpiration stream, and diffused outward from the veins to the epidermal cell walls within minutes of reaching a particular level in the leaf. We conclude that the suberized mestome sheath of rye leaves is freely permeable to solutes moving apoplastically through radial primary cell walls.     

Cytotaxonomic study ofChelidonium majus L. s. l.

The present paper deals with cytotaxonomy ofChelidonium majus L. s. 1. taxa and their hybrids. Based on results of hybridization experiments, cytology and reproductive isolation, a new combination,Ch. asiaticum (Hara) Krahulcová, is proposed. The structure of the aggregate is as follows:Ch. majus L. subsp.Majus (2n=12, distributed in Europe),Ch. majus L. subsp.grandiflorum (DC.)Printz (2n=12, S. Siberia, China) andCh. asisaticum (2n=10, E. Asia). Karyotypes ofCh. m. subsp.grandiflorum andCh. asiaticum are compared in detail.     

Comparative substrate-inhibitor analysis of mink liver monoamine oxidases

Comparative substrate-inhibitor analysis of catalytic properties of liver monoamine oxidases (MAO) was performed in the mature males of the American mink Mustela vison and the European mink Mustela lutreola. The action on the MAO activity was studied of alkaloids of the benzo[c]phenanthridine group: sanguinarine and chelidonine, diisoquinoline alkaloid berberine, medicinal agents “Ukrain” and “Sanguirythrin” as well as derivatives of 2-propylamine: deprenyl and chlorgylin. The latter turned out to be irreversible inhibitor of the MAO A form, whereas deprenyl-irreversible inhibitor of the MAO B form in both studied mink species. The selectivity of action of each inhibitor on the corresponding liver MAO form for the species M. vison was one order of magnitude stronger than for the species M. lutreola. All studied alkaloids as well medicinal agents on their basis have been shown to be specific irreversible inhibitors of the intermediate strength of the liver MAO A form of both mink species. They inhibit the enzymatic deamination of serotonin, tyramine, and tryptamine without affecting the deamination reaction of benzylamine and β-phenylethylamine (at concentrations of 10 mM and lower). Out of five studied isoquinoline agents, the medication “Ukrain” and alkaloid chelidonine have the highest inhibitory action; the agent “Sanguirythrin” and alkaloids berberine and sanguinarine produce the weaker monoamine oxidase effect. The revealed specificity of action of the studied inhibitors is an indirect evidence for the presence in the liver enzymes of both mink species, like in the rat liver enzyme, of several molecular forms.     

Effects of seed aggregation on the removal rates of elaiosome-bearing Chelidonium majus and Viola odourata seeds carried by Formica polyctena ants

In a field dominated by Formica polyctena Foerst. ants, we examined the effect of seed aggregation on the seed-removal rates of two plant species: a large-seeded obligate myrmecochore Viola odorata L. and a small-seeded diplochore Chelidonium majus L., which was autochorous as well as myrmecochorous. The effect was statistically non-significant in V. odorata but significant in C. majus, with more closely aggregated seeds having higher removal rates. The large seeds of the obligate myrmecochore were more quickly discovered and repeatedly removed by ant workers than were the small seeds of the diplochore.     

Osmotic and specific effects of magnesium sulphate on the mineral content ofZea mays

Summary The effects which the osmotic pressures 2.0, 3.5 and 5.0 atm., obtained by the addition of either magnesium sulphate or PEG-4000 to the standard nutrient solution, have on the development and mineral content ofZea mays var. INIA 8302 are assessed. The osmotic effect causes a greater absorption of cations, potassium being the element most readily absorbed. The specific effect of magnesium sulphate causes a decrease in the total of anions and in the water content of the plant and partly offsets the decrease which the osmotic effect produces in the fresh weight.     

Isolation of an Antibacterial Substance from Mahonia fortunei and its Biological Activity against Xanthomonas oryzae pv. oryzicola

This study aimed to isolate the antibacterial substance from Mahonia fortunei and determine its antibacterial activity against Xanthomonas oryzae pv. oryzicola (Xoc). Bacterial leaf streak of rice, caused by Xoc, is an important rice disease and difficult to control. During a screening of antibacterial plants against plant pathogenic bacteria at an early stage, the extract from M. fortunei was found to have a strong inhibitory effect on Xoc. In this study, the chemical components of M. fortunei stems were extracted using methanol solvent, the antibacterial substance was isolated and purified by liquid–liquid partition and silica gel column chromatography and its structure was identified by nuclear magnetic resonance. The effect and mode of action of the antibacterial substance on bacterial leaf streak of rice were also detected under greenhouse conditions. Two compounds were identified, berberine and jatrorrhizine, which had a strong inhibitory effect on Xoc. The antibacterial activity of berberine was stronger, with a half‐maximal inhibitory concentration (IC₅₀) of 2.9008 mg/l. At the concentration of 0.5 g/l, its control efficacy on bacterial leaf streak of rice was more than 84%. Additionally, berberine could be absorbed by rice leaves and be translocated up and down in the rice plant, and the effective period was long, but its capability of lateral translocation inside the blade was poor.