Development of fertilized ovules and their role in the growth of the pea pod

The histological development of fertilized ovules during fruit-set and development in pea ( Pisum sativum L. cv. Alaska) has been investigated. Killing the ovules on day 0 (anthesis) or day 1 prevented fruit-set and resulted in ovary degeneration. When the ovules were destroyed at later stages the ovaries developed, though the rate of growth of the pod was reduced significantly. Pollination in pea occurs normally the day before anthesis, and fertilization of the egg cell 32 to 48 h later. The first divisions of the zygote and endosperm nuclei started simultaneously (ca 48 h after pollination) but the endosperm developed more rapidly than the embryo; the embryo sac cavity was lined with free endosperm nuclei at the time of beginning suspensor elongation. Extracts of endosperm and ovule coats from ovules at day 7 after anthesis showed fruit-set activity in pea, the latter material having about 3 times more activity than the former per ovule basis. These results indicate that fertilization of the ovule is necessary for fruit-set in pea, and that compounds which induce fruit-set are probably synthesized in the ovules following fertilization.     

The roles of veratryl alcohol and nonionic surfactant in the oxidation of phenolic compounds by lignin peroxidase

Veratryl alcohol (VA) at higher concentration stimulated the lignin peroxidase (LiP)-catalyzed oxidation of phenolic compounds remarkably. This novel phenomenon was due to its competition with the phenols for the active site of the enzyme and to the high reactivity of the formed cation radical of VA (VA+*) which resulted in an additional oxidation of the phenols. The influence of the nonionic surfactant Tween 80 on the VA-enhanced LiP-catalyzed oxidation of phenols depended on its concentration. At lower concentration it had a small synergetic effect but at higher concentration it decreased the initial rate. Studies of the capillary electrophoretic behavior of LiP in the presence of Tween 80 showed that this effect was caused by the surfactant aggregation on LiP which, at higher surfactant concentrations, might impede the access of VA to its binding site on LiP and, consequently, the VA+* formation.     

Evaluation of Phytochemical Compositions and Biological Properties of Achillea gypsicola at Different Phenological Stages

Phytochemicals, which are commonly found at different levels in many medicinal plants, are natural strong antioxidants used in traditional medicine. In this research, determination of differences of phytochemical compositions and biological properties were aimed as periodically (pre‐, full and post flowering) and daily (6 am, 1 pm and 8 pm) in Achillea gypsicola Hub.‐Mor . The volatile oils belonging to A. gypsicola were obtained by hydrodistillation and analyzed by gas chromatography‐flame ionization detection (GC‐FID) and gas chromatography‐mass spectrometry (GC/MS). The antimicrobial activities of the volatile oils were determined with disc diffusion method. The microdilution method was used to determine minimum inhibitory concentration (MIC). Total phenolic and flavonoid contents were determined by spectrophotometric methods and antioxidant capacities were evaluated by 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH) free radical, reducing power (RP) and metal chelating activity (MCA) assay. In addition, the phenolic acid and flavonoid compositions were evaluated by reversed phase‐high‐performance liquid chromatography (RP‐HPLC). This study presented a comprehensive report for the first time on evaluation of the phytochemical composition and the biological properties of A. gypsicola at different phenological stages. Thirty‐two compounds, containing the major component as camphor, 1,8‐cineole and borneol, were detected. Designated harvest time for the highest yield of volatile oils was found to be at full flowering stage‐1 pm. It has been observed that the volatile oil composition changes periodically and even daily. Also, in this research, menthol and menthone were found as the composition of volatile oil in Achillea species for the first time. Full flowering stage was found as the richest period in terms of phenolic acid and flavonoid compositions of A. gypsicola for the first time. The species examined in this research showed a high antioxidant and antimicrobial activity in comparison to other studies with Achillea species. The volatile oils exhibited high performances with range of inhibition zones (8.3–42.3 mm) and minimum inhibitory concentration values (2.25—144 μg/ml). Besides, a high correlation between antioxidant activity and phenolic content of A. gypsicola was found. These results suggest that A. gypsicola can be used as a safe source in the cosmetic, food and pharmaceutical industries.     

紫背金盘(Ajuga nipponensis)次生物质对桔全爪螨(Panonychus citri)的驱避作用

研究了紫背金盘Ajuga nipponensis Makino各溶剂提取物和部分化合物对桔全爪螨Panonychus citri McGregor雌成螨及其产卵的驱避作用.结果表明,石油醚萃取物、乙酸乙酯萃取物具有较强的生物活性.在0.1 g · L-1时, 石油醚和乙酸乙酯萃取物对该螨处理1d后的产卵忌避率分别为:84.86%、69.88%;2d后为89.49%、82.19%;对雌成螨驱避率分别为:85.08%、68.66%;2d后为50.96%、69.84%.乙酸乙酯萃取物经分离得到四类化合物,结果表明:馏分Ⅰ为长链脂肪酸混合物,具有较强生物活性,2000μg/ml和1000μg/ml处理1d后,产卵忌避率分别为:80.77%、74.77%;2d后为73.81%、72.59%.2000μg/ml处理1d后对雌成螨的驱避率为:69.88%;2d后为74.24%.刺槐素Ⅱ、新克罗烷化合物Ⅲ和β-蜕皮甾酮Ⅳ在2000μg/ml均不表现活性.对馏分Ⅰ中的4个主要化合物单体进行活性测定,结果表明:十六烷酸、十六烷酸甲酯、十六烷酸乙酯和十八烷酸甲酯在2000μg/ml处理时,1d后,产卵驱避率分别为:75.18%、61.76%、59.18%和66.49%;2d后产卵驱避率为:66.67%、31.15%、46.75%和44.84%;雌成螨驱避率分别为:1d后,67.53%、63.79%、59.26%和68.00;2d后,67.23%、43.96%、48.23%和64.19%.在1000μg/ml处理时,1d 后,产卵驱避率分别为:59.21%、59.16%、57.02%和61.40%;1d后,雌成螨驱避率分别为:69.64%、61.43%、55.76%和64.00%.     

Initial steps of phloroglucinol metabolism in Penicillium simplicissimum

The pathway for the aerobic catabolism of 1,3,5-trihydroxybenzene (phloroglucinol) by a new strain of Penicillium was investigated using both in vivo and in vitro cell-free systems. The fungal strain was isolated by enrichment on phloroglucinol and identified as P. simplicissimum (Oud) Thom. It grew optimally at pH 5.5 and 27°C with 119 mM (1.5%w/v) of phloroglucinol in a basal mineral salts medium. Vapours of the crystalline substrate placed in a Petri-plate lid supported the growth of the fungal colonies on the agar surface. Mycelia grown on phloroglucinol accumulated 1,2,4-trihydroxybenzene and resorcinol in the medium. Washed, resting mycelia grown on phloroglucinol, when resuspended in a buffer utilized oxygen in the presence of catechol, resorcinol, pyrogallol and phloroglucinol. A NADPH-dependent reductase in the cell-free extract reduced phloroglucinol to dihydrophloroglucinol. This electron donor could not be replaced by NADH. Resorcinol hydroxylase, phloroglucinol reductase, catechol-1,2-oxygenase, and catechol-2,3-oxygenase were detected in cell-free extracts of mycelia grown on phloroglucinol. The possible steps in the degradation of phloroglucinol are discussed.     

Survey of aliphatic glucosinolates in Sicilian wild and cultivated Brassicaceae

In the frame of the activities carried out to exploit Sicilian local cultivars of brassicas, we focused our attention on some of the potential health compounds of various local cruciferous crops. These compounds are of interest to improve the quality of the produce with the aim to develop new cultivars capable of providing functional foods able to prevent disease. In this context, we surveyed for the presence of specific glucosinolates in local cultivars of broccoli, cauliflower, kale, and in some wild species widespread in Sicily, using as control various commercial cultivars. Glucosinolate composition varied extensively among species and crops of the same species, such as cauliflower, broccoli and kale. Cultivar variation for glucosinolate profile was also observed for some crops. For example, Sicilian cultivars of cauliflower possessing colored curds displayed a high content of glucosinolates, glucoraphanin in particular, compared to white curd commercial cultivars. Also some wild species had a high content of other glucosinolates.     

不同产地杜仲根际土真菌群落结构的差异性分析

基于宏条码技术对采自贵州遵义县、湖南慈利县和四川旺苍县3个杜仲产地根际土壤的真菌群落组成进行了分析。研究获得根际土壤真菌有效序列共10 775条,聚类后OTUs对应的真菌类群涉及子囊菌门Ascomycota、担子菌门Basidiomycota、接合菌门Zygomycota、球囊菌门Glomeromycota、壶菌门Chytridiomycota、类原生动物门Rozellomycota的22纲、59目、103科、108个属的550个真菌分类单元(OTUs)。在属的水平上,湖南慈利样本(ECS)有119个属,优势属为镰刀菌属Fusarium,相对丰度为16.31%,其次是被孢霉属Mortierella(14.67%)、毛壳菌科Chaetomiaceae一未定属(13.01%)和木霉属Trichoderma(11.37%)等。四川旺苍样本(EWS)共有116个属,优势属为丝齿菌属Hyphodontia,相对丰度32.32%,其次为镰刀菌Fusarium(13.41%)等。贵州遵义样本(EZS)有110个属,优势属为被孢霉属Mortierella,相对丰度30.74%,其次是木霉属Trichoderma(25.96%)和镰刀菌属Fusarium(4.85%)等。α多样性分析表明,不同产地杜仲根际土壤真菌群落组成有明显差异。遵义样本(EZS)、慈利样本(ECS)和旺苍样本(EWS)的Shannon多样性指数从高到低为:ECS(2.7661)> EZS(2.4841)>EWS(2.1326)。不同杜仲产区土壤理化因子及4个主要活性成分(松脂醇二葡萄糖苷、绿原酸、京尼平苷酸和京尼平苷)与根际土真菌群落组成的相关性分析表明,4个主要活性成分皆为四川旺苍(EWS)产地含量最高,其次是湖南慈利,贵州遵义最低甚至不含京尼平苷。杜仲根际土壤样品真菌物种组成丰富,不同地区杜仲根际土壤真菌群落组成、分类单元的相对丰度及优势分类单元皆存在不同程度差异。杜仲根际土真菌群落结构的多样性与土壤理化因子及杜仲主要有效成分含量具有一定相关性。杜仲根际土真菌群落组成的结构谱是杜仲道地性的重要特征。     

Permanent Stained Preparations of Synovial Fluid for Detection of Calcium Compounds Using Alizarin Red S

Permanent preparations of air dried synovial fluids were prepared by staining calcium compounds with alizarin red S stain; each slide was coverslipped with Permount. Variables studied were: (a) concentration of the solution of alizarin red S, (b) pH of staining solution, (c) time of incubation in staining solution and aqueous and ethanolic content of staining solution. The staining effect of each solution was tested on calcium pyrophosphate dihydrate, calcium oxalate, apatite and monosodium urate (MSU). Of all the solutions, best results were obtained with 0.25% alizarin red S in 50% ethanol at pH 7.0 for 30 min. With this solution, the calcium-containing compounds were well stained. MSU did not stain and still preserved negative birefringence on polarizaton. Fixation of smears with ethanol served a double purpose: It fixed the slides without dissolving or removing MSU or the calcium compounds, yet it did dissolve five corticosteroids commonly used for intra-articular injection which may interfere with interpretation of compensated polarized light microscopy of synovial fluids.     

The hidden face of food phenolic composition

Plant polyphenols are extremely diverse, due to the occurrence of several basic structures, numerous substitutions and, for some groups, of polymers (tannins). Plant polyphenol composition depends on the plant species and organ, with some molecules specific of particular plant families while others are ubiquitous. The polyphenol content is classically assessed by global analysis methods, which lack specificity and accuracy. These methods have been replaced with high performance liquid chromatography (HPLC), that enables accurate determination of individual molecules, provided they can be unambiguously identified and calibration curves can be established. However, HPLC analysis is restricted to simple compounds and difficult to apply in the case of complex extracts. Further difficulties encountered in the case of polymers include irreversible adsorption on the stationary phases. Proanthocyanidin analysis by HPLC after acid-catalysed depolymerisation in the presence of a nucleophile permits to overcome these problems and shows that proanthocyanidins predominate in the polyphenol composition of most plants. Large varietal differences in tannin quantitative and qualitative composition were observed for all plant species studied. Moreover, analysis is usually performed after extraction, which may lead to significant underestimation of the polyphenol content, since a large proportion is not extracted by usual solvents. This may be due to covalent binding to other plant constituents and to non-covalent adsorption on plant solids. Such matrix effect also influences the taste perception of polyphenols and their fate in the digestive tract, from in-mouth interactions with salivary proteins to their metabolism by colon microflora, with potential influence on bioavailability.