Diterpenes from Salvia broussonetii transformed roots and their insecticidal activity

The new diterpenes brussonol (1) and iguestol (6alpha,11-dihydroxy-12-methoxy-abieta-8,11,13-triene) (2) with an icetexane and a dehydroabietane skeleton, respectively, have been isolated from hairy root cultures of Salvia broussonetii. Other previously known diterpenes, 7-oxodehydroabietane, 11-hydroxy-12-methoxyabietatriene, taxodione, inuroyleanol, ferruginol, deoxocarnosol 12-methyl ether, cryptojaponol, pisiferal, sugiol, isomanool, 14-deoxycoleon U, 6alpha-hydroxydemethylcryptojaponol, demethylsalvicanol, and demethylcryptojaponol, were also obtained from these roots. The insect antifeedant and toxic effects of several of these compounds were investigated against the insect pests Spodoptera littoralis and Leptinotarsa decemlineata. Additionally, their comparative cytotoxic effects were tested on insect Sf9 and mammalian CHO cells. Demethylsalvicanol (4) was a moderate antifeedant to L. decemlineata, whereas brussonol (1) was inactive. 14-Deoxycoleon U (15) was the strongest antifeedant, whereas demethylcryptojaponol (11) was toxic to this insect. None of these compounds had antifeedant or negative effects on S. littoralis ingestion or weight gains after oral administration. Demethylcryptojaponol (11) was cytotoxic to mammalian CHO and insect Sf9 cell lines, followed by the icetexane derivative brussonol (1), with moderate cytotoxicity in both cases. The remainder of the test compounds showed a strong selective cytotoxicty to insect Sf9 cells, with demethylsalvicanol (4) being the most active.     

Volatile components and key odorants of fennel (Foeniculum vulgare Mill.) and thyme (Thymus vulgaris L.) oil extracts obtained by simultaneous distillation-extraction and supercritical fluid extraction

Volatile oil extracts of fennel seeds (Foeniculum vulgare Mill.) and thyme leaves (Thymus vulgaris L.) were obtained by simultaneous distillation-extraction (SDE) and supercritical fluid extraction (SFE) and analyzed by gas chromatography-mass spectrometry (GC-MS). In general, fennel oil extracted by SDE and SFE showed similar compositions, with trans-anethole, estragole, and fenchone as the main components. In contrast, thymol and p-cymene, the most abundant compounds in thyme leaves, showed big differences, with generally higher amounts of monoterpenes obtained by SDE. However, in this case, the differences between the extracts were higher. Key odorants of fennel seeds determined by gas chromatography-olfactometry (GC-O) showed similar patterns when applying SDE and SFE. trans-Anethole (anise, licorice), estragole (anise, licorice, sweet), fenchone (mint, camphor, warm), and 1-octen-3-ol (mushroom) were the most intense odor compounds detected in fennel extracts. Thymol and carvacrol, with oregano, thyme, and spicy notes, were identified as key compounds contributing to the aroma of thyme leaves.     

Polyphenol oxidase from Dominga table grape

Dominga grape polyphenol oxidase (PPO) was extracted using phase partitioning with Triton X-114. The enzyme was obtained in latent state and could be optimally activated by the presence of 0.2% sodium dodecyl sulfate (SDS) at pH 6.0. In the absence of SDS, the enzyme showed maximum activity at acid pH. The kinetic parameters of the enzyme at pH 3.0 and 6.0 in the presence of SDS were calculated. The effect of several inhibitors was studied, tropolone being the most effective with a K(i) value of 18 muM. The effect of cyclodextrins was also studied, and the complexation constant K(c) between G(2)-beta-cyclodextrins and 4-tert-butylcatechol was calculated using the enzymatic method (K(c) = 13960 M(-)(1)). The evolution of the color parameters (L, a, b) of liquefied grape berries was inhibited by inhibitors of PPO activity, such as diethyldithiocarbamate, metabisulfite, and G(2)-beta-cyclodextrins, indicating that enzymatic browning by PPO is the main process involved in the browning of Dominga grape juice at room temperature.     

Real-time polymerase chain reaction based assays for quantitative detection of barley, rice, sunflower, and wheat

Quality assurance is a major issue in the food industry. The authenticity of food ingredients and their traceability are required by consumers and authorities. Plant species such as barley (Hordeum vulgare), rice (Oryza sativa), sunflower (Helianthus annuus), and wheat (Triticum aestivum) are very common among the ingredients of many processed food products; therefore the development of specific assays for their specific detection and quantification are needed. Furthermore, the production and trade of genetically modified lines from an increasing number of plant species brings about the need for control within research, environmental risk assessment, labeling/legal, and consumers' information purposes. We report here the development of four independent real-time polymerase chain reaction (PCR) assays suitable for identification and quantification of four plant species (barley, rice, sunflower, and wheat). These assays target gamma-hordein, gos9, helianthinin, and acetyl-CoA carboxylase sequences, respectively, and were able to specifically detect and quantify DNA from the target plant species. In addition, the simultaneous amplification of RALyase allowed bread from durum wheat to be distinguished. Limits of detection were 1 genome copy for barley, sunflower, and wheat and 3.3 copies for rice real-time PCR systems, whereas limits of quantification were 10 genome copies for barley, sunflower, or wheat and approximately 100 haploid genomes for rice real-time PCR systems. Real-time PCR cycling conditions of the four assays were stated as standard to facilitate their use in routine laboratory analyses. The assays were finally adapted to conventional PCR for detection purposes, with the exception of the wheat assay, which detects rye simultaneously with similar sensitivity in an agarose gel.     

Effect of accelerated autolysis of yeast on the composition and foaming properties of sparkling wines elaborated by a champenoise method

Five mutants (obtained by UV mutagenesis) and the parent strain were selected to produce sparkling wines following the traditional or champenoise method. The wines were aged with the yeast for 9 months, with samples being taken each month for analytical and sensory determinations. The wines elaborated with mutant strain IFI473I demonstrated an accelerated release of protein, amino acids, and polysaccharides. An analysis of the secreted polysaccharides revealed that mannose was the major sugar present. The effects of the products released by yeasts on the foaming properties of the wines were determined by both sensory and instrumental analysis. In all cases, the wines elaborated with mutant strain IFI473I showed improved foaming properties as compared to wines fermented without this strain. Similar results were obtained at a decreased aging time of 6 months, thereby confirming the capacity of IFI473I strain to carry out an accelerated autolysis. These results demonstrate that mutant strain IFI473I can significantly reduce production times of high-quality sparkling wines.     

Illinois Soil Nitrogen Test Method Optimization for Analysis of Temperate Mineral Grassland Soils

ABSTRACT

A reliable and practical test that can provide timely measurements of the levels of mineralizable nitrogen (MN) in soil is critical for improving the accuracy of N fertilizer applications for grassland and crops. The Illinois soil N test (ISNT) is considered to be a good estimate of MN, once soils are grouped according to soil characteristics such as the drainage type and sampling depth. To date, development and evaluation of the ISNT method has been conducted using arable soils mainly in North America where, in general, soils have lower levels of soil organic matter (SOM) compared to temperate grassland soils. We evaluated the effects of two pre-treatment soil aggregate sizes of <1 mm and <2 mm on the yield and recovery of MN (1) across temperate grassland soil types, and (2) across a 6-h interval diffusion period. No significant difference existed in the concentrations of ISNT-N between the two soil aggregate sizes of each soil type. For both aggregate sample sizes, the recovery of spiked amino sugar-N glucosamine from a temperate grassland soil was generally linear until hour 5, after which the quantities of recovered N diminished. Although N recovery after 6 h of diffusion at 50°C (±1°C) was less than 100% in both aggregate size samples, the response models indicated that the standard ISNT protocol using a 5-h diffusion period is appropriate for temperate grassland soils. The incomplete recovery of N in these mineral soils suggested that the protocol could be further optimized for temperate soils with high organic matter content and additional evaluation of the temperature during diffusion within an enclosed environment may be required using N (spiked glucosamine-N) recovery studies.     

Changes in broccoli (Brassica oleracea L. Var. italica) health-promoting compounds with inflorescence development

Changes in phenolic compounds, total glucosinolates, and vitamin C were monitored during the productive period along five inflorescence development stages of three broccoli commercial cultivars (Marathon, Monterrey, and Vencedor). In an attempt to identify differences due to agronomic factors, broccoli cultivars were grown under different sulfur fertilization with poor (15 kg/ha) and rich (150 kg/ha) rates. Phenolic compounds and vitamin C concentrations showed, in all broccoli cultivars, a rising trend from the first stage until the over-maturity stage, both for rich and poor sulfur fertilization. Significant differences were detected in the first two stages between rich and poor sulfur fertilization in total glucosinolates for all broccoli cultivars, where the highest concentration was always observed in the second development stage (used as minimally processed product) during poor fertilization. With regard to the last three stages, the glucosinolate concentration in the poor sulfur fertilization started to slope down until the over-maturity stage. Where rich sulfur fertilization is concerned, the highest level was reached during the third stage (used as minimally processed product also), and after that, glucosinolate concentration decreased until the fifth stage.     

Sulfluramid volatility reduction by beta-cyclodextrin

Sulfluramid is an expensive active principle of insecticidal baits that is lost by volatilization during the pelletization of baits. To increase the thermal stability of sulfluramid, we tested its molecular encapsulation in beta-cyclodextrin (beta-CD), using molar ratios of 1:1 and 1:2 (sulfluramid:beta-CD), using the complex preparation techniques of coprecipitation and kneading. The physical mixture of sulfluramid and beta-CD was also tested for comparison. The products of complexation were characterized by differential scanning calorimetry, thermogravimetry, and derivative thermogravimetry, indicating the formation of a sulfluramid/beta-CD complex and showing that the release of the complexed sulfluramid occurs in the range of 270-300 degrees C, a temperature range that is well above the temperature at which sulfluramid sublimates (40 degrees C). This result warrants a reduced sulfluramid loss in the preparation of insecticidal baits. The preparation of the complex by kneading with molar ratio of 1:2 gave the highest yield of complex, about 64%, in relation to the theoretical maximum.