Effect of selected phytotoxins from Guanomyces polythrix on the calmodulin-dependent activity of the enzymes cAMP phosphodiesterase and NAD-kinase

The effect of a series of phytotoxins isolated from the fungus Guanomyces polytrix on calmodulin (CaM)-dependent nicotinamide adenine dinucleotide kinase (NADK) and CaM-dependent cyclic nucleotide phosphodiesterase (PDE) activities was investigated. The results indicated that (2S,3S)-5-hydroxy-6,8-dimethoxy-2,3-dimethyl-4H-2,3-dihydronaphtho[2,3-b]-pyran-4-one, (2S,3S)-5-hydroxy-6,8,10-trimethoxy-2,3-dimethyl-4H-2,3-dihydro-naphtho[2,3-b]-pyran-4-one, (2S,3R)-5-hydroxy-6,8-dimethoxy-2,3-dimethyl-2,3-dihydro-4H-naphtho[2,3-b]-pyran-4-one, (2S,3R)-5-hydroxy-6,8,10-trime-thoxy-2,3-dimethyl-2,3-dihydro-4H-naphtho[2,3-b]-pyran-4-one, 5-hydro-xy-6,8-dimethoxy-2,3-dimethyl-4H-naphtho[2,3-b]-pyran-4-one, rubrofusarin B, and ergosta-4,6,8(14),22-tetraen-3-one inhibited the activation of both target enzymes in the presence of CaM. On the other hand, (2S)-5-hydroxy-6,8-dimethoxy-2-methyl-4H-2,3-dihydronaphtho[2,3-b]-pyran-4-one and (2S)-5-hydroxy-6,8,10-trimethoxy-2-methyl-4H-2,3-dihydronaphtho-[2,3-b]-pyran-4-one inhibited the activation of PDE and the basal activity of NADK. Thus, these phytotoxins are CaM inhibitors and may exert their phytotoxic action by inhibiting the CaM-dependent process, although they could also interfere with other cellular metabolic phenomena. This is the first report of the use of the NADK assay to detect or quantify CaM inhibitors, and it could be a valuable tool for studying those CaM isoforms regulating NADK.     

Phosphorus and liming effects on early growth of selected plant species grown on an ultisol

Abstract

A series of pot experiments were conducted to test the effect of phosphorus (P) application and liming on early growth performance of Zea mays, Mucuna pruriens and nine woody species (Albizia ferruginea, Cajanus cajan, Dactyladenia barteri, Dalbergia sissoo, Enterolobium cyclocarpum, Flemingia macrophylla, Gliricidia sepium, Leucaena leucocephala, and Pentacletra macrophylla) on an Ultisol. Low reponse to liming and high response to P application were observed in this trial. Early top growth of the eleven species was correlated to seed size and seed nitrogen (N), P, and calcium (Ca) contents. Highest correlation was observed with seed P content (r = 0.896^**). Large seeded size species (D. barteri, E. cyclocarpum, M. pruriens, and P. macrophylla) showed the least reponse to P application, while the remaining small seeded size species showed significant responses to P application. The large seeded species also showed relatively less increases in N, P and Ca uptakes with P application and liming. The pot test can be used in prescreening of woody species for use on low P and acid soils. The results also emphasize the need for seed size consideration in multipurpose trees and shrubs selection for establishment in low P and on degraded soils     

Inhibitory effects of lignans on the activity of human matrix metalloproteinase 7 (matrilysin)

Inhibitory effects of nine dibenzylbutyrolactone lignans on a human matrix metalloproteinase, matrilysin, were examined. All of the lignans examined inhibited matrilysin with the IC(50) values ranging from 50 to >280 microM. Matairesinol, which has the basic structure of the other lignans, showed the weakest inhibition. Lignans with methylenedioxy ring(s) or a hydroxyl group at the C5-position inhibited matrilysin more strongly than matairesinol. 5-Hydroxypluviatolide, which has both a methylenedioxy ring and a hydroxyl group at the C5-position, was the most potent inhibitor (IC(50) = 50 microM), suggesting that the introduction of these two elements might enhance synergistically the inhibitory activity of lignans. 5-Hydroxypluviatolide inhibited matrilysin in a competitive manner, and its inhibitory effect was greatly suppressed by the presence of another competitive inhibitor, dimethyl sulfoxide. The precursors of matairesinol, coniferyl alcohol and secoisolariciresinol, had no inhibitory activity, indicating that the dibenzylbutyrolactone structure is essential for the inhibition. It has been shown that lignans have the potential to inhibit matrilysin, and the knowledge of their structure-activity relationship might be beneficial to developing selective inhibitors for matrix metalloproteinases.     

A mixture of grass and clover combines the positive effects of both plant species on selected soil biota

The introduction of N₂-fixing white clover (Trifolium repens) in grassland is a management measure that may contribute to sustainable grassland systems by making them less dependent on inorganic fertilizers. However, little is known about the impact of this measure on soil biota and ecosystem services. We investigated earthworms, nematodes, bacteria and fungi in an experiment in which white clover-only and a mixture of grass and white clover without fertilization were compared with grass-only with and without fertilization.In comparison with grass-only, white clover-only had a lower total root biomass and a lower C/N-ratio in the above- and below-ground plant biomass. These plant characteristics resulted in a lower bacterial biomass, a lower fungal biomass, a higher proportion of bacterivorous nematode dauerlarvae, a lesser proportion of herbivorous nematodes and a greater abundance of earthworms in clover-only.The quantity and quality (C/N-ratio) of the above- and below-ground plant biomass in the mixture of grass and white clover (20–30% clover in the DM) was comparable with grass fertilized with 150 kg N ha⁻¹ of inorganic fertilizer. Differences between these treatments might show specific clover effects in the grass–clover mixture on soil biota other than quantity and C/N-ratio of the litter. However, the only differences were a higher proportion of bacterivorous nematode dauerlarvae and a different nematode community composition in grass–clover.The soil structure in white clover-only showed a higher proportion of angular blocky elements, a lower penetration resistance, a higher number of earthworm burrows, a higher potential N-mineralization and respiration than the soil in grass-only. This suggests that clover stimulates the ecosystem services of water infiltration and supply of nutrients, but is less conducive to soil structure maintenance. The grass–clover mixture differed from grass-only in a higher respiration and from clover-only in a higher percentage of soil crumbs. We suggest that when clover is introduced in grassland to reduce the reliance on inorganic fertilizer, the mixture of grass and clover maintains the positive impact of grass roots on soil structure and increases the supply of nutrients via the soil food web. Thus, a grass–clover mixture combines the agronomic benefits of the two plant types.     

Evaluation of selected enzymes for thiamine determination

Seven commercially available enzymes were studied for suitability as substitutes in the AOAC thiamine determination, because the enzyme Takadiastase used in the method has not been available since 1976, and alternative enzymes were likewise unavailable or unsuitable for releasing thiamine from its phosphate esters. Four factors (substrate ester, enzyme level, time/temperature, and pH) at 2 levels were studied in a 2(4) factorial arrangement of treatments. Data were expressed in terms of mean percentage conversion (MPC) and were statistically evaluated by analysis of variance. Significant main effects and any interactions among treatments were calculated. Takadiastase and alpha-amylase (Miles) with MPCs of 101 and 102, respectively, appeared effective in dephosphorylation within method parameters. Potato phosphatase appeared marginally suitable. Wheat germ phosphatase, alpha-amylase (Sigma), Mylase 100, and Clarase 40,000 were judged unacceptable as enzyme substitutes.     

植物精油海藻酸钠复合膜对双孢蘑菇的抑菌保鲜效果研究

为了研究植物精油对双孢蘑菇贮藏期主要腐败菌的抑制效果,该文以新鲜双孢蘑菇为试验材料,分离、纯化并鉴定引起双孢蘑菇贮藏期腐败的主要病原菌;通过体外抑菌活性试验,筛选出抑菌效果较好的植物精油,并将此精油应用于复合膜的保鲜试验中,验证其抑菌保鲜效果。通过对双孢蘑菇主要致腐微生物进行分离、纯化得到9株单菌落,经致病性试验得到主要病原真菌为3号菌,对该病原菌18S rDNA序列进行进化树分析,确定其为丰孢木霉菌(Trichoderma sp.)。选用姜黄、连翘、桉叶、当归4种精油对3号菌菌株进行抑菌活性试验,筛选出较优精油种类及浓度。结果表明,桉叶油对丰孢木霉菌的抑制效果最好,其次是连翘油,姜黄油和当归油无明显抑制作用。桉叶油最小抑菌浓度(minimum inhibitory concentration, MIC)为0.312 5μL/mL,最小杀菌浓度(minimum bactericidal concentration, MBC)为0.625μL/mL;连翘油MIC为0.625μL/mL,MBC为1.25μL/mL。通过对海藻酸钠/桉叶油复合膜和海藻酸钠单一膜的性能指标进行测定和对比发现,桉叶油的添加可显著提高膜的气体阻隔性能和机械性能。分别用海藻酸钠/桉叶油复合膜、海藻酸钠单一膜和普通PE膜对双孢蘑菇进行保鲜试验,结果表明,添加桉叶油的复合膜可有效降低双孢蘑菇子实体的衰老和致病腐烂,具有显著的抑菌保鲜效果。该研究结果可为双孢蘑菇贮藏期防腐和天然杀菌保鲜剂的选择提供一定的理论依据。     

Inhibitory effects of the total and water-soluble concentrations of nine different metals on the dehydrogenase activity of a loess soil

 This study focuses on a comparison of the microbial toxicity of nine metals, including As as a metalloid and two species of Cr. A loess soil [Ap horizon, clay 15.2%, organic C 1.12%, pH(CaCl₂) 7.02] was spiked with 8–12 geometrically increasing doses of the metals. The dehydrogenase assay (2-p-iodophenyl-3-p-nitrophenyl-5-phenyltetrazoliumchloride method) was combined with sorption and solubility experiments. The resulting dose-response curves and sorption isotherms were used to derive total doses that caused definite percentage inhibitions [i.e. effective doses (ED) causing a 10–90% reduction in dehydrogenase activity (dha)] as well as the corresponding toxic solution concentrations causing the same reductions in dha (i.e. effective concentrations; EC₁₀–EC₉₀). Based on total doses, the toxicity decreased in the following order with ED₅₀ values (mg kg^–1) given in brackets: Hg (2.0)>Cu (35)>Cr(VI) (71)>Cr(III) (75)>Cd (90)>Ni (100)>Zn (115)>As (168)>Co (582)>Pb (652). With regard to solution concentrations, toxicity decreased in the order (EC₅₀ in mg l^–1): Hg (0.003)>Pb (0.04)>Cu (0.05)>Cd (0.14)>Zn (0.19)>Cr(III) (0.62)>Ni (0.69)>Co (30.6)>As (55.5)>Cr(VI) (78.1). The retention of the metals by the soil differed strongly. Pb, Cu, and Hg exhibited the highest and Ni, As, and Cr(VI) the lowest sorption constants (Freundlich K values: 2455, 724, 348, 93, 13, and 0.06 mg kg^–1, respectively). The sorptivity of the metals and their microbial toxicity in the aqueous phase were characteristically related: metals with a strong toxic action in the soil solution were adsorbed by the soil to a high degree and vice versa. Therefore, especially for metals with a high inherent toxicity, sorption is an effective way of immobilizing them and temporarily detoxifying soil. Received: 2 July 1998     

Application of analytical standards in method development: selected chlorinated phenols and herbicides

Pure analytical standards are key components in the successful development of accurate analytical methods. When such compounds are unavailable commercially, a laboratory synthesis activity can provide such materials for use in method development. An example of such an approach is the synthesis of ethyl ethers of chlorinated phenols and ethyl esters of several phenoxy herbicides. Individual synthesis steps and the characterization of such materials are described. The use of such analytical standards in method development is vital for defining method characteristics. In addition, such materials are helpful in diagnosing chromatographic and mass spectral problems in the course of this work. Key mass spectral information including parent and daughter scans obtained from these analytical standards by using tandem mass spectrometry provide the basis for a new analytical method with increased sensitivity and specificity.     

芳香族化合物对土壤硝化作用的抑制研究

Aromatic compounds (ACs) in soil can induce competitive inhibition for soil NH₃ oxidation, and nitrification inhibitors can be used to this end. A laboratory incubation experiment was performed with 12 nitroaromatic compounds (NACs), 15 amidoaromatic compounds (AACs) and 20 hydroxyaromatic compounds (HACs) to assess the inhibitory effects of ACs on soil nitrification. Based on these results, the critical and optimal concentrations of ACs were determined for better inhibitory effects. Most of the test ACs were able to inhibit soil nitrification; the effectiveness differed with soil type. Among the ACs, the NACs with m-nitryl, amino or hydroxyl and the AACs with a nitro group or a chlorine atom on aromatic ring or with a p-hydroxyl were more effective. 3-nitroaniline, 4-aminophenol and 3-nitrophenol showed the greatest potential as nitrification inhibitors. The critical concentration of these compounds in brown soil and cinnamon soil was found to be 0.5 mg kg^-1 soil. Due to the toxicity, carcinogenicity and mutagenicity of ACs, further toxicological and ecotoxicological research is necessary before ACs are used as nitrification inhibitors in agricultural and horticultural practices.     

Inhibitory effects of Zingiber officinale Roscoe derived components on aldose reductase activity in vitro and in vivo

Ginger (Zingiber officinale Roscoe) continues to be used as an important cooking spice and herbal medicine around the world. Scientific research has gradually verified the antidiabetic effects of ginger. Especially gingerols, which are the major components of ginger, are known to improve diabetes including the effect of enhancement against insulin-sensitivity. Aldose reductase inhibitors have considerable potential for the treatment of diabetes, without increased risk of hypoglycemia. The assay for aldose reductase inhibitors in ginger led to the isolation of five active compounds including 2-(4-hydroxy-3-methoxyphenyl)ethanol (2) and 2-(4-hydroxy-3-methoxyphenyl)ethanoic acid (3). Compounds 2 and 3 were good inhibitors of recombinant human aldose reductase, with IC50 values of 19.2 +/- 1.9 and 18.5 +/- 1.1 microM, respectively. Furthermore, these compounds significantly suppressed not only sorbitol accumulation in human erythrocytes but also lens galactitol accumulation in 30% of galactose-fed cataract rat model. A structure-activity relationship study revealed that the applicable side alkyl chain length and the presence of a C3 OCH3 group in the aromatic ring are essential features for enzyme recognition and binding. These results suggested that it would contribute to the protection against or improvement of diabetic complications for a dietary supplement of ginger or its extract containing aldose reductase inhibitors.     

Inhibitory effects of Orostachys japonicus extracts on the formation of N-nitrosodimethylamine

In Korea, Orostachys japonicus has been used traditionally as a drug and health food. The aim of this study was to investigate possible inhibitory effects of O. japonicus extracts on the formation of N-nitrosodimethylamines (NDMA). Chloroform extraction was the most effective method for recovering the highest number of phenolic compounds and flavonoids; in these extracts the greatest nitrite-scavenging activity and inhibition of NDMA formation occurred at pH 2.5. The chloroform extract was separated into 10 fractions (J1-J10); fraction J4 inhibited NDMA formation by 90.1 +/- 0.4%. This fraction was then separated into five subfractions (J4-1-J4-5) using a silica gel column. Subfractions J4-2 [(+)-catechin] and J4-4 (3,4-dihdroxybenzoic acid) inhibited NDMA formation by 89.5 +/- 0.9 and 77.6 +/- 0.8%, respectively.     

Inhibitory effects of SO2 on biofiltration of aldehydes

Biofiltration is a suitable method for eliminating volatile biodegradable organic chemical from waste gases. However, if substances inhibiting microbiological activity are present in the waste gas, biofiltration efficiencies will be poor. Results are presented on inhibition of biofiltration by SO₂. Inhibition was observed first in a pilot scale experiment on odor abatement in a food processing factory. Then it was shown in laboratory scale experiments that SO₂. concentrations less than 10 ppm have no measurable effect. A concentration of 40 ppm results in 60% inhibition but original efficiency can be restored when the SO₂ supply is shut off. A concentration of 100 PPM of SO₂ results in complete and permanent inhibition.     

Effects of selected trace metals on germinating seeds of six plant species

Seeds of cabbage, lettuce, millet, radish, turnip, and wheat were treated with solutions containing Be, Ni, Tl, or V, and subsequent effects on seed germination and radicle elongation were measured after three days. Treatment with low concentrations of Be, Ni, or V stimulated root elongation in most species. Higher concentrations of these elements and all treatment with Tl caused reductions in root elongation. In general, turnip and lettuce were the most sensitive of the plants studied to the metals tested, while wheat and millet were the least sensitive.     

Nitrogen effects on total flavonoids,chlorogenic acid,and antioxidant activity of the medicinal plant Chrysanthemum morifolium

Chrysanthemum morifolium (Ramat.) has a long history of cultivation and use as a traditional medicine and tea plant in China. A greenhouse experiment with potted soil–quarz mixture studied the effects of nitrogen supply (0, 56, 112, 167, 224, 334, 501, 556, and 668 mg N kg^–1) on concentrations and ratios of total flavonoids and chlorogenic acid in the flowers of C. morifolium using spectrophotometric and HPLC methods. The antioxidant activity of the flowers was determined as the radical scavenging activities of hydroxyl, superoxide anion, and 2,2‐diphenyl‐1‐picrylhydrazyl hydrate (DPPH) free radicals. A high N supply decreased the concentrations of total flavonoids by 18%–35% and that of chlorogenic acid by 8%–60% compared to a low N‐supply rate. At the same time, increasing N supply significantly decreased the antioxidant activity of the flowers. The antioxidant activity of C. morifolium flowers was significantly positively correlated with the concentrations of total flavonoids and chlorogenic acid. We conclude that an N supply in excess of 300 mg (kg soil)^–1 will negatively affect the antioxidant activity and thereby reduce the quality of C. morifolium flowers.     

Determination of halogenated phenols in raw and potable water by selected ion gas chromatography-mass spectrometry

Pentafluorobenzylation and in situ acetylation are compared in the determination of phenol and halogenated phenols in water samples. The latter technique is considered superior to the former for determining phenols at the ng/L level because of less background interference and better recoveries (80% or better except for pentachlorophenol and trichloroguaiacol which had recoveries of about 60%). Further evaluation of the in situ technique by electron capture gas chromatography and gas chromatography-mass spectrometry shows that the latter, in the selected ion monitoring mode, is more suitable because, unlike GC-ECD, it can confirm and quantitate all phenols. In particular, GC-ECD could not detect even high levels of phenol and the monohalogenated phenols. Phenols at 5-473 ng/L were detected in some Canadian drinking water supplies by the in situ acetylation technique combined with GC-MS.     

Interactive effects of warming, soil humidity and plant diversity on litter decomposition and microbial activity

Human activity has induced a multitude of global changes that are likely to affect the functioning of ecosystems. Although these changes act in concert, studies on interactive effects are scarce. Here, we conducted a laboratory microcosm experiment to explore the impacts of temperature (9, 12 and 15 °C), changes in soil humidity (moist, dry) and plant diversity (1, 4, 16 species) on soil microbial activity and litter decomposition.We found that changes in litter decomposition did not mirror impacts on microbial measures indicating that the duration of the experiment (22 weeks) may not have been sufficient to determine the full magnitude of global change effects. However and notably, changes in temperature, humidity and plant litter diversity/composition affected in a non-additive way the microbial parameters investigated. For instance, microbial metabolic efficiency increased with plant diversity in the high moisture treatment but remained unaffected in low moisture treatment suggesting that climate changes may mask beneficial effects of biodiversity on ecosystem functioning. Moreover, litter decomposition was unaffected by plant litter diversity/composition but increased with increasing temperature in the high moisture treatment, and decreased with increasing temperature in the low moisture treatment.We conclude that it is inevitable to perform complex experiments considering multiple global change agents in order to realistically predict future changes in ecosystem functioning. Non-additive interactions highlight the context-dependency of impacts of single global change agents.     

Effect of clay minerals on the activity of polysaccharide cleaving soil enzymes

Kinetic analyses of the influence of montmorillonite, palygorskite and kaolinite on the activity of three polysaccharide cleaving soil enzymes revealed an inhibitory effect that is rather due to the structure than to chemical characteristics of the clays. The fibrous palygorskite was found to be highly effective towards the starch cleaving α-amylase and amyloglucosidase, whereas the layer silicates caused only a slight inhibition. Interactions with cellulase were affected by sorption of cellulose by montmorillonite and kaolinite. When the enzyme was allowed to be sorbed by the clays before cellulose was added, a clay-enzyme complex is formed, which in contrast to the starch cleaving enzymes remains active. Compared to the native enzyme, the bound enzyme is not stabilized with respect to temperature, but the optimum pH is found to be at pH 6 instead of pH 4.8.     

水分胁迫对苹果属植物抗氧化酶活性的影响研究

试验研究水分胁迫对 3种苹果属植物湖北海棠、西府海棠和新疆海棠根系和叶片抗氧化酶活性的影响结果表明 ,随水分胁迫的加重 ,3种海棠抗氧化酶活性逐渐增强 ,当土壤绝对含水量降至 6 .5 %时 ,过氧化氢酶 (CAT)活性首先达高峰 ,而超氧化物歧化酶 (SOD)活性高峰则出现于土壤绝对含水量为 2 .8%时 ,之后 2种酶活性逐渐降低 ,复水后又均有所升高 ;过氧化物酶 (POD)活性高峰出现于土壤绝对含水量为 3.2 %时 ,之后逐渐下降 ,至复水后仍呈下降趋势 ;丙二醛 (MDA)含量随水分胁迫的加重而逐渐增加 ,当土壤绝对含水量降至 1 .8%时其含量最高 ,复水后则呈下降趋势。种间抗氧化能力耐旱型新疆海棠 >西府海棠 >干旱敏感型湖北海棠     

紫外辐射与复合胁迫对植物抗氧化酶系统的影响

植物抗氧化酶系统对UV-B辐射增强与复合胁迫的响应是植物抵御逆境伤害的关键机制之一。结合近年来国内外研究成果,概述了UV-B辐射对植物抗氧化酶基因表达、转录与翻译的影响,植物抗氧化酶系统对UV-B辐射与干旱、臭氧、高温、盐渍、重金属及酸雨复合胁迫的响应,以及CO2、抗坏血酸、α-萘乙酸、硒、激光辐射和稀土等调节因子对UV-B辐射下植物抗氧化酶系统的作用。