落地生根内生固氮菌多样性和促生特性

【背景】内生固氮菌可以定殖于植物体内为植物提供营养物质,还能通过代谢促进植物生长,目前对于落地生根内生菌的研究鲜见报道。【目的】研究落地生根中内生固氮菌多样性。【方法】从表面消毒的植物组织中分离纯化内生菌,并通过乙炔还原法测定菌株的固氮酶活性。采用SDS-PAGE全细胞蛋白电泳和IS指纹图谱对菌株聚类,各类群代表菌株进行16S rRNA基因系统发育分析和生理生化鉴定。测定菌株固氮、分泌生长素和ACC脱氨酶、产铁载体、溶磷和解钾等促生特性。【结果】从落地生根中分离纯化出26株内生固氮菌,聚为5个类群,隶属于4个属的5个菌种,且各类群代表菌株具有多种促生功能。【结论】从落地生根中分离获得的内生菌具有丰富的遗传多样性和促生特性,并且存在新的微生物资源,有待开发利用。     

In vitro efficacy of Bryophyllum pinnatum leaf extracts as potent therapeutics

Leaf extracts of Bryophyllum pinnatum (BPEs) are used in several countries. Contextually, evaluation of the therapeutic potential of these was carried out in this study to explore antioxidant and antityrosinase potential through different in vitro methods. The radical scavenging properties of BPEs were studied using various techniques, based on the 1,1-diphenyl-2-picrylhydrazyl (DPPH) dot blot thin-layer chromatography (TLC) method, electron paramagnetic resonance (EPR) spectroscopy, metal chelation, β-carotene bleaching, inhibition of DNA breakage on agarose gel, and lipid peroxidation inhibition using liver and brain microsomes. EC₅₀ values of the results reflected that aqueous-methanolic BPE was the most active one. Antibrowning potential of the fresh leaf extract showed an antityrosinase property, with EC₅₀ values of enzymatic assay of tyrosinase inhibitory activity further advocating the findings.     

Effects of aqueous leaf extract of Bryophyllum pinnatum on guinea pig tracheal ring contractility

Aqueous leaf extract of Bryophyllum pinnatum Lam (Crassulaceae) is used as a cough remedy and for the prophylaxis of asthma. Since drugs used for the prophylaxis of asthma may be acting on airway smooth muscles, we investigated the effects of aqueous leaf extract of the plant on the contractile responses of isolated tracheal rings. Guinea pigs were grouped into non-sensitized, ovalbumin (OA)-sensitized, OA-sensitized but 200 mg/kg/day x 21 extract-treated, and OA-sensitized but 400 mg/kg/day x 21 extract-treated. The extract was administered orally. Tracheal rings obtained from the four groups were mounted in organ baths and used to test spasmolytic and antispasmodic effects of the extract on histamine or carbachol-induced contractions. Concentrations of 0.125-1.0 mg/ml of the extract did not relax histamine or carbachol-induced precontractions. The presence of 0.25-1.0 mg/ml of the extract in organ baths significantly reduced the maximal contractile responses (Emax) to cumulative concentrations of histamine or carbachol irrespective of the experimental group. pD2 values were significantly reduced for histamine and carbachol in rings obtained from 400 mg/kg/day x 21 extract-treated group. It is concluded that aqueous leaf extract of B. pinnatum possesses antispasmodic effects on the guinea pig tracheal rings. The results lend credence to the use of the extract for the prophylaxis of asthma in ethnomedicine.Keywords: Bryophyllum pinnatum; Tracheal rings; Anti-asthmatic; Antispasmodic; Herbal medicine.     

Radical scavenging activity of ribonuclease inhibitor from cow placenta

Cow placenta ribonuclease inhibitor (CPRI) has been purified 5062-fold by affinity chromatography, the product being homogeneous by sodium dodecyl sulfate-gel electrophoresis. The chemiluminescence technique was used to determine the radical scavenging activities of CPRI toward different reactive oxygen species (ROS) including superoxide anion (O2-*), hydroxyl radical (OH*), lipid-derived radicals (R*), and singlet oxygen (1O2). CPRI could effectively scavenge O2-*, OH*, R*, and 1O2 at EC50 of 0.12, 0.008, 0.009, and 0.006 mg/ml, respectively. In addition, the radical scavenging activities of CPRI were higher than those of tea polyphenols, indicating that CPRI is a powerful antioxidant.     

中华补血草多酚提取物对自由基的清除能力

采用超声波辅助浸提—大孔树脂吸附法从中华补血草[ Limonium sinense (Girard) Kuntze]根、根茎、叶和花中获得多酚提取物,并对它们的得率和组成成分进行了初步分析;在此基础上,比较研究了不同部位多酚提取物对DPPH·、·OH和O2的清除能力.实验数据表明:中华补血草根、根茎、叶和花多酚提取物的得率分别为12.42％、5.98％、5.27％和3.98％,差异明显;多酚提取物中总酚、原花色素、黄烷醇和总黄酮含量变化幅度较大,分别为51.87％ ～61.60％、5.62％～39.47％、3.69％～12.46％和2.53％ ～35.97％;其中,总酚、原花色素和黄烷醇含量均以根多酚提取物最高,总黄酮含量以花多酚提取物最高,均与其他部位多酚提取物有显著差异.随质量浓度提高,4个部位多酚提取物对3种自由基的清除率总体上逐渐增大;其中根多酚提取物对DPPH·的清除作用最强,半数清除质量浓度(ρSC50)为38.52 μg·L-1,显著低于阳性对照芦丁和BHT(ρSC50分别为67.40和74.25 μg·L-1);花多酚提取物对·OH和O2的清除能力最强,ρSC50分别为53.51和74.00 μg·L-1,均低于芦丁;总体上,4个部位多酚提取物对DPPH·的清除能力由强至弱依次为根、根茎、叶、花,对·OH和O2的清除能力由强至弱依次为花、根、叶、根茎.结果显示:中华补血草不同部位多酚提取物均具有较强的自由基清除能力,其中,根和花多酚提取物中高含量的原花色素和黄酮类成分分别与其自由基清除能力密切相关.     

Radical scavenging properties of polyamines

Di- and polyamines are effective scavengers of free radicals generated in a number of chemical and in vitro enzyme systems. Free radical production was quantified spectrophotometrically using nitroblue tetrazolium and cytochrome c or by electron spin resonance. Levels of superoxide radical formed either enzymatically with xanthine oxidase or chemically from riboflavin or pyrogallol were significantly inhibited by spermine, spermidine, putrescine and cadaverine at 10 and 50 mM. The more reactive hydroxyl radical generated by the Fenton reaction was also effectively scavenged by di- and polyamines. In addition, the production of superoxide radical by senescing microsomal membranes was inhibited by di- and polyamines, as was the superoxide-dependent conversion of 1- aminocyclopropane-1-carboxylic acid (ACC) to ethylene. The efficacy of polyamine-scavenging appears to be correlated with the extent of amination suggesting the involvement of amino groups. It is also apparent that some of the physiological effects of polyamines, in particular their propensity to inhibit lipid peroxidation and retard senescence, may be attributable to their radical-scavenging capability.     

Radical scavenging properties of genistein

The reactivity of genistein toward reactive radical species has been investigated by means of pulse radiolysis. The values of rate constants, respectively 2.3 x 10(10) M(-1)s(-1) and 1.3 x 10(10) M(-1)s(-1) for the reaction with hydroxyl radical at pH 8.3 and 3.0, are close to diffusion limit indicating that genistein is a potent hydroxyl radical scavenger. The reactivity of genistein towards one-electron oxidants has also been investigated. The rate constants k = 4.6 x 10(9) M(-1)s(-1) (pH 8.3) and 6.7 x 10(8) M(-1)s(-1) (pH 7.6) have been determined for the reaction of genistein with *N3 and Br2*- radicals, respectively. For both oxidants the rate constants at pH 3 does not exceed 10(8) M(-1)s(-1). The differences in reactivity of genistein towards the oxidants at different acidity of the solution have been assumed to arise from the acid-base equilibria of genistein. The dissociation constants for genistein (pKa: 7.2, 10.0, and 13.1) have been evaluated spectroscopically. The influence of acid-base equilibria on bond dissociation energy and ionization potential for genistein has also been investigated by means of DFT calculations. It has been concluded on the basis of these calculations that monoanionic form of genistein existing at physiological pH is more powerful radical scavenger than the neutral molecule.     

Radical scavenging activities of niacin-related compounds

We investigated whether niacin-related compounds had radical-scavenging activity by electron spin resonance methods. Many compounds, but not trigonelline, had radical-scavenging activity against hydroxyl radicals. However, for the nitric oxide radical and 1,1-diphenyl-2-picrylhydrazyl radical, only nicotinic acid hydrazide and isonicotinic acid hydrazide had scavenging activities. These results suggest that the moiety of hydrazide might have an important role in scavenging abilities of various radicals.     

Radical scavenging activity of protein from tentacles of jellyfish Rhopilema esculentum

In this study, radical scavenging activity of protein from tentacles of jellyfish Rhopilema esculentum (R. esculentum) was assayed including superoxide anion radical and hydroxyl radical scavenging. The protein samples showed strong scavenging activity on superoxide anion radical and values EC50 of full protein (FP), first fraction (FF), second fraction (SF), and 30% (NH4)2 SO4 precipitate (Fr-1) were 2.65, 7.28, 1.10, and 22.51 microg/mL, respectively, while values EC50 of BHA, BHT, and alpha-tocopherol were 31, 61, and 88 microg/mL, respectively. Also, the protein samples had strong scavenging effect on hydroxyl radical and the values EC50 of FP, FF, SF, Fr-1, and Fr-2 were 48.91, 27.72, 1.82, 16.36, and 160.93 microg/mL, but values EC50 of Vc and mannitol were 1907 and 4536 microg/mL, respectively. Of the five protein samples, SF had the strongest radical scavenging activity and may have a use as a possible supplement in the food and pharmaceutical industries. The radical scavenging activity was stable at high temperature so that R. esculentum may be used as a kind of natural functional food.     

A reevaluation of the peroxynitrite scavenging activity of some dietary phenolics

Peroxynitrite is implicated in many diseases. Hence, there is considerable interest in potential therapeutic peroxynitrite scavengers. Diet-derived phenolics have been claimed to be powerful peroxynitrite scavengers. However, the reactivity of peroxynitrite can be significantly modified by bicarbonate and this has not been considered in evaluations of the scavenging activity of phenols. Bicarbonate (25 mM) significantly decreased the ability of several phenolic compounds (caffeic acid, o- and p-coumaric acid, gallic acid, ferulic acid) but not others (catechin and epicatechin) to inhibit peroxynitrite-mediated tyrosine nitration. Bicarbonate (25 mM) also decreased the ability of catechin, epicatechin, quercetin and ferulic acid but not chlorogenic acid, gallic acid, caffeic acid and o-coumaric acid to inhibit peroxynitrite-mediated alpha(1)-antiproteinase inactivation. These results show that physiological concentrations of bicarbonate substantially modify the ability of dietary phenolics to prevent peroxynitrite-mediated reactions. When assessing compounds for peroxynitrite scavenging, experiments should be conducted in the presence of bicarbonate to avoid misleading results.     

Allelopathic potential of the phenolics from the roots of Pluchea lanceolata

Aqueous leachates of roots of the perennial weed Pluchea lanceolata (DC.) C. B. Clarke, its root-incorporated soil and rhizosphere soil, interfered with the seedling growth of certain plant species. The soils from the rhizosphere zone of this plant had significantly higher total phenolics and HPLC analysis revealed that phenolic fractions represented by retention times of 1.6, 1.9, 2.5 (simple phenol, chlorogenic acid and phloroglucinol respectively), 3.7 and 4.3 min were contributed by roots of the weed to the soil. The phenolic fraction represented by the retention time 3.3 (formononetin 7-O-glucoside) was detected in the weed's rhizosphere soils and not in the rootincorporated soils. UV spectral studies established the presence of phloroglucinol, simple phenol, chlorogenic acid, formononetin 7-O-glucoside, and methylated coumarins in the root leachate, which affect the seedling growth of mustard ( Brassica juncea ). Present research established the allelopathic potential of P. lanceolata roots, and the possible involvement of allelopathy in its interference success.     

Anti-inflammatory and antinociceptive potential of major phenolics from Verbascum salviifolium Boiss

The potential effects of flavonoids, phenylethanoid and neolignan glycosides from the aerial parts of Verbascum salviifolium Boiss. were studied in the p-benzoquinone-induced writhing reflex, for the assessment of the antinociceptive activity, and in carrageenan- and PGE1-induced hind paw edema and 12-O-tetradecanoyl-13-acetate (TPA)-induced ear edema models in mice, for the assessment of the anti-inflammatory activity. Through bioassay-guided fractionation and isolation procedures ten compounds from the aqueous extract of the plant, luteolin 7-O-glucoside (1), luteolin 3'-O-glucoside (2), apigenin 7-O-glucoside (3), chrysoeriol 7-O-glucoside (4), beta-hydroxyacteoside (5), martynoside (6), forsythoside B (7), angoroside A (8), dehydrodiconiferyl alcohol-9'-O-beta-D-glucopyranoside (9) and dehydrodiconiferyl alcohol-9-O-beta-D-glucopyranoside (10), were isolated and their structures were elucidated by spectral techniques. Results have shown that 1, 2, 3 and 5 significantly inhibited carrageenan-induced paw edema at a 200 mg/kg dose, while 1, 2 and 5 also displayed anti-inflammatory activity against the PGE1-induced hind paw edema model. However, all the compounds showed no effect in the TPA-induced ear edema model. The compounds 1 and 2 also exhibited significant antinociceptive activity.     

Radical scavenging activities of alpha-alanine C60 adduct

Water-soluble alpha-alanine C60 adduct was synthesized, and its scavenging abilities for superoxide anion O2- and hydroxyl radical *OH were studied by the spectrophotometry and chemiluminescence. It was found that alpha-alanine C60 adduct showed an excellent efficiency in eliminating superoxide anion and hydroxyl radical. The 50% inhibiting concentration (IC50) for superoxide was 184 microg/mL by spectrophotometry and 292 microg/mL by chemiluminescence. The IC50 for hydroxyl radical was 42 microg/mL. In different test systems, the results showed that alpha-alanine C60 adduct had comparable radical scavenging abilities as thiourea and ascorbic acid, and was proved to be an effective scavenger for superoxide anion and hydroxyl radical. It can be prospected that water-soluble alpha-alanine C60 adduct will be useful in radical related biomedical fields.     

Radical scavenging ability of some compounds isolated from Piper cubeba towards free radicals

The purpose of this study was to identify the antioxidant activity of 16 compounds isolated from Piper cubeba (CNCs) through the extent of their capacities to scavenge free radicals, hydroxyl radical (HO^?), superoxide anion radical () and 2,2‐diphenyl‐1‐picrylhydrazyl radical (DPPH^?), in different systems. Electron paramagnetic resonance (EPR) and 5,5‐dimethyl‐1‐pyrroline‐N‐oxide, DMPO, as the spin trap, and chemiluminescence techniques were applied. Using the Fenton‐like reaction [Fe(II) + H₂O₂], CNCs were found to inhibit DMPO? OH radical formation ranging from 5 to 57% at 1.25 mmol L^?1 concentration. The examined CNCs also showed a high DPPH antiradical activity (ranging from 15 to 99% at 5 mmol L^?1 concentration). Furthermore, the results indicated that seven of the 16 tested compounds may catalyse the conversion of superoxide radicals generated in the potassium superoxide/18‐crown‐6 ether system, thus showing superoxide dismutase‐like activity. The data obtained suggest that radical scavenging properties of CNCs might have potential application in many plant medicines. Copyright © 2010 John Wiley & Sons, Ltd.     

Radical scavenging and NO-releasing properties of selected beta-adrenoreceptor antagonists

It is claimed that novel beta-adrenolytic drugs possess superior antioxidant properties as compared to classical selective or non-selective beta-adrenoceptor antagonists. Here we tested this notion by analyzing radical scavenging properties of selected beta-adrenolytic drugs and their ability to release nitric oxide in biological preparations. Selective beta1-adrenolytics such as nebivolol, atenolol, metoprolol and non-selective beta-adrenolytics with alpha1-receptor blocking properties such as carvedilol and labetalol were chosen for analysis. NO-releasing properties of nebivolol and carvedilol distinguished third generation beta-adrenolytics from their older counterparts while the reactivity towards hydroxyl and peroxyl radicals discerns only carvedilol but not nebivolol. Thus, superior clinical efficacy of third generation beta-adrenolytics may be related to their ability to release NO rather then to their direct antioxidant properties.     

Radical scavenging activities of Heracleum aquilegifolium Wight (Apiaceae) fruit oils in vitro

The fruits of Heracleum aquilegifolium Wight (Apiaceae) were collected from Western Ghats of the Indian Peninsula. The essential oils were extracted by hydrodistillation. The chemical composition of the essential oils was analysed by gas chromatography and gas chromatography-mass spectrometry (GC-MS). Beta-Pinene (22.3%), 1,8-cineole (20.3%), and beta-phellandrene (12.4%) were the main components of H. aquilegifolium fruit oils. The antioxidant properties of essential oils of H. aquilegifolium were examined by different procedures namely reducing power ability, 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity, nitric oxide radical scavenging activity, hydrogen peroxide scavenging activity, hydroxyl radical scavenging activity, superoxide anion scavenging activity, and metal chelating activity. The antioxidant activities were compared with those of synthetic antioxidants and standard drugs such as butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), ascorbic acid, alpha-tocopherol, curcumin, and quercetin. The study confirmed the possible antioxidant potential of essential oils tested with various in vitro antioxidant methods. The presence of monoterpenes in combination with other components in the oils could be responsible for the activity.     

Radical scavenging activity and oxidative modification of citrus dehydrin.

Dehydrins are ubiquitous proteins produced by plants in response to water stress. Their functions, however, are not fully understood. The overexpression of Citrus unshiu Marcov. dehydrin (CuCOR19) enhanced cold tolerance in transgenic plants by reducing lipid peroxidation promoted by cold stress, suggesting that the CuCOR19 protein directly scavenges radicals. In this paper, we report the radical scavenging activity and oxidative modification of CuCOR19. The hydroxyl radical generated by the Fe2+/H2O2 system and peroxyl radical generated from 2, 2'-azobis (2-amidinopropane) (AAPH) were scavenged by CuCOR19, but hydrogen peroxide and superoxide were not. The scavenging activity for the hydroxyl radical and peroxyl radical of CuCOR19 was more potent than that of mannitol, and approximately equal to that of serum albumin, which is known as an antioxidative protein in mammals. CuCOR19 was degraded by the hydroxyl radical and peroxyl radical in a time- and dose-dependent manner. Mannitol and thiourea inhibited the degradation. Analysis of the amino acid composition of CuCOR19 indicated that glycine, histidine, and lysine, which are major residues in many dehydrins, were targeted by the hydroxyl radical. These results suggest that CuCOR19 is a radical scavenging protein, and may reduce oxidative damage induced by water stress in plants.