Cloning of a novel L-amino acid oxidase from Trichoderma harzianum ETS 323 and bioactivity analysis of overexpressed L-amino acid oxidase

L-amino acid oxidases (L-AAOs) have been isolated from many organisms, such as snake, and are known to have antibacterial activity. To the best of the authors' knowledge, this is the first report of the cloning of cDNA encoding a novel Trichoderma harzianum ETS 323 L-amino acid oxidase (Th-L-AAO). The protein was overexpressed in Escherichia coli and purified to homogeneity. Comparisons of its deduced amino acid sequence with the sequence of other L-AAOs revealed the similarity to be between 9 and 24%. The molecular mass of the purified protein was 52 kDa, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The enzyme substrate specificity was highest for L-phenylalanine, and its optimal pH and temperature for activity were 7 and 40 °C, respectively; exogenous metal ions had no significant effect on activity. Circular dichroism spectroscopy indicated that the secondary structure of Th-L-AAO is composed of 17% α-helices, 28% β-sheets, and 55% random coils. The bacterially expressed Th-L-AAO also mediated antibacterial activity against both gram-positive and gram-negative food spoilage microorganisms. Furthermore, a three-dimensional protein structure was created to provide more information about the structural composition of Th-L-AAO, suggesting that the N-terminal sequence of Th-L-AAO may have contributed to the antibacterial activity of this protein.     

Antidepressant effect of GABA-rich monascus-fermented product on forced swimming rat model

γ-Aminobutyric acid (GABA) has several well-known physiological functions including antihypertension and antidepression. In this research, we focus on the antidepressant effects of oral administration of GABA-rich Monascus-fermented product in depression animal model (forced swimming test, FST) by Sprague-Dawley rats, and try to find its possible mechanism in the brain monoamine system. GABA and the Monascus-fermented product (MFP) significantly decreased the duration of immobility time in a short-term test. In a long-term test, the antidepressant-like effect of MFP was better than that of GABA at the same dosage (2.6 mg/kg), and the efficacy of MFP was similar to that of fluoxetine. Moreover, GABA might recover the level of monoamines norepinephrine, dopamine (DA), and 5-hydroxytryptamine (5-HT) in hippocampus and normalize the turnover ratio of 5-HT and DA in hippocampus and amygdala. In addition to the functions of GABA, the MFP has more potential in decreasing the turnover ratio of DA in the frontal cortex and striatum to improve depressive symptoms.     

柑桔黄龙病热治疗的研究

柑桔黄龙病村有“均匀黄化型”和“斑驳型”两类型的症状。两类型的橙类和蕉柑苗木经49℃湿热空气处理50～60分钟或用50℃处理50分钟,89株苗木全部恢复健康,经58～60个月后全未发病,而对照的病苗种后其病情日趋严重,陆续死亡。试验结果除进一步表明热处理可以消灭黄龙病病原类立克次体外,并进一步表明“黄化型”和“斑驳型”的病原有可能属于同一种病原或相近似的病原。     

Organochlorine pesticide residues in Italian citrus essential oils, 1991-1996

Organochlorine pesticide contamination in 148 lemon essential oils, 123 sweet orange oils, 121 mandarin oils, and 147 bergamot oils produced in Italy in the years 1991-1996 was studied by HRGC-ECD. Confirmation analyses were carried out by GC-MS. Tetradifon, dicofol and its decomposition product 4,4'-dichlorobenzophenone were found. Over the course of the study dicofol and tetradifon residues steadily decreased; the percentage of contaminated samples reflects this course and decreases considerably from 1991 to 1996.     

Antibacterial activity of Cuminum cyminum L. and Carum carvi L. essential oils

Essential oils extracted by hydrodistillation from fruits of Cuminum cyminum L. and Carum carvi L. were analyzed by gas chromatography (GC) and GC-mass spectrometry (MS). The main components of C. cyminum oil were p-mentha-1,4-dien-7-al, cumin aldehyde, gamma-terpinene, and beta-pinene, while those of the C. carvi oil were carvone, limonene, germacrene D, and trans-dihydrocarvone. Antibacterial activity, determined with the agar diffusion method, was observed against Gram-positive and Gram-negative bacterial species in this study. The activity was particularly high against the genera Clavibacter, Curtobacterium, Rhodococcus, Erwinia, Xanthomonas, Ralstonia, and Agrobacterium, which are responsible for plant or cultivated mushroom diseases worldwide. In general, a lower activity was observed against bacteria belonging to the genus Pseudomonas. These results suggest the potential use of the above essential oils for the control of bacterial diseases.     

Determination of imidazolidine-2-thione (ethylenethiourea) in ethylenebisdithiocarbamate formulations

Zineb was most seriously contaminated with imidazolidine-2-thione (ethylenethiourea; ETU), followed by mancozeb, and maneb. The type of formulation also affects ETU generation and a wettable powder produced ETU more easily than a flowable formulation. The ETU contents of the ethylene-bisdithiocarbamate (EBDC) mixed fungicides formulations were erratic. Low ETU contents were found in EBDC mixed fungicides, even though some samples had been stored in the laboratory for five years. The cupric ion and the N, O-containing group of other fungicides might have a stabilizing effect on the degradation of EBDC to ETU. The survey indicated that EBDC mixed formulations could be a useful means of suppressing ETU generation in commercial products.     

Genetic improvement effects on yield stability in durum wheat genotypes grown in Italy

In durum wheat (Triticum durum Desf.) the improvement of yield stability represents an important component for agricultural progress worldwide. This work reports on the evaluation of the yield performance of 65 durum wheat genotypes (landraces, old and new cultivars with different years of release and advanced breeding lines), in a range of Italian environments to assess the changes in adaptation and yield stability achieved over the last century as results of the genetic improvement. The Additive Main effect and Multiplicative Interaction (AMMI) analysis was used to capture a large portion of the Genotype × Environment interaction (GE) sum of squares and to separate main and interaction effects. The breeding strategies adopted during the last decades have contributed to reduce the interaction of genotypes with environments selecting genotypes with better stability across a wide range of locations and years and modern genotypes outperformed the old ones in all test environments with a strong adaptability to improved fertility. The old cultivars were characterized by a minimal responsiveness to improved environmental conditions, showing an almost stable nominal yield in agreement with the concept of “biological” or “static” stability. In contrast, the modern cultivars were highly responsive to fertility improvements and showed a pronounced adaptation to high-input environments. Notably, within the group of modern cultivars two of them, Tiziana and Giusto, showed a high-yield stability in biological terms and a high nominal yield across the tested environments. Giusto and Tiziana were the best cultivars in terms of both nominal yield and minimal GE interaction, indicating that selecting for improved yield potential may increase yield in a wide range of environments.