Antifungal effects of different organic extracts from Melia azedarach L. on phytopathogenic fungi and their isolated active components

Extracts from different parts of Melia azedarach L. were studied as potential antifungal agents for selected phytopathogenic fungi. In a serial agar dilution method, hexanic and ethanolic extracts from fruit, seed kernels, and senescent leaves exhibited fungistatic activity against Aspergillus flavus,Diaporthe phaseolorum var. meridionales, Fusarium oxysporum, Fusarium solani, Fusarium verticillioides, and Sclerotinia sclerotiorum. Both hexanic extract from senescent leaves and ethanolic extract from seed kernel were highly effective on all tested fungi, with minimum inhibitory concentration (MIC) values ranging from 0.5 to 25 mg/mL and 0.5 to 5 mg/mL, respectively. In addition, all of the above-mentioned extracts showed fungicidal activity on these fungi, with ethanolic seed kernel extract being the most active. Three compounds displaying activity against F. verticillioides were isolated from the ethanolic seed kernel extract and were characterized as vanillin (1), 4-hydroxy-3-methoxycinnamaldehyde (2), and (+/-)-pinoresinol (3), with MICs of 0.6, 0.4, and 1.0 mg/mL, respectively. These compounds also showed a synergistic effect when combined in different concentrations, needing four times less concentration to reach complete inhibition in the growth of F. verticillioides.     

Antifeedant and insecticide properties of a limonoid from Melia azedarach (Meliaceae) with potential use for pest management

In the course of screening for novel naturally occurring insecticides from plants, the activity of the fruit extract of the Argentinian Melia azedarach L. (Meliaceae) and its recently described limonoid meliartenin were investigated. The antifeedant activity of the fruit extract was tested on a variety of herbivore and granivorous insects through choice tests. Sixteen of 17 species belonging to three orders consume significantly less food when treated with the extract. The bioactivity of the isolated active compound meliartenin and its interchangeable isomer 12-hydroxiamoorastatin (1) was further studied. In choice tests, compound 1 inhibited feeding of Epilachna paenulata Germ. (Coleoptera, Coccinellidae) larvae, with an ED(50) value of 0.80 microg/cm(2), comparable to that of azadirachtin (2) and lower than that of toosendanin (3) (0.72 and 3.69 microg/cm(2), respectively), both compounds used for comparison purposes. In no-choice tests, E. paenulata larvae reared on food treated with 1 or 2 ate less, gained less weight, and suffered greater mortality rates than control larvae. The activity of compound 1 was comparable to that of 2, with LD(50) values of 0.76 and 1.24 microg/cm(2), respectively, at 96 h. Shorter LT(50) values were recorded for 1 at 4 and 1 microg/cm(2) in comparison with 2. Thus, M. azedarach fruit extract and its active principle have interesting potential for use in pest control programs.     

Protein from red cabbage (Brassica oleracea) seeds with antifungal, antibacterial, and anticancer activities

A 30 kDa antifungal protein was purified from red cabbage ( Brassica oleracea ) seeds. It exhibited a molecular mass and N-terminal amino acid sequence disinct from those of previously isolated Brassica antifungal proteins. The protocol used entailed ion exchange chromatography on Q-Sepharose and SP-Sepharose followed by fast protein liquid chromatography on Mono S. The protein hindered mycelial growth in Mycosphaerella arachidicola (with an IC50=5 μM), Setospaeria turcica, and Bipolaris maydis. It also inhibited the yeast Candida albicans with an IC50=96 μM. It exerted its antifungal action by permeabilizing the fungal membrane as evidenced by staining with Sytox green. The antifungal activity was stable from pH 3 to 11 and from 0 to 65 °C. It manifested antibacterial activity against Pseudomonas aeruginosa (IC50=53 μM). Furthermore, after 48 h of culture, it suppressed proliferation of nasopharyngeal cancer and hepatoma cells with IC50=50 and 90 μM, respectively.     

Terpendole M, a novel indole-diterpenoid isolated from Lolium perenne infected with the endophytic fungus Neotyphodium lolii.

Terpendole M (1), a novel indole-diterpenoid, was isolated from perennial ryegrass (Lolium perenne) infected with the endophytic fungus Neotyphodium lolii. It was identified as 14alpha-hydroxyterpendole C by NMR and mass spectral techniques. The known indole-diterpenoids paspaline (3) and 13-desoxypaxilline (4) were also isolated from perennial ryegrass for the first time. Terpendole M was less tremorgenic than terpendole C (2) in a standard mouse bioassay. These findings provide clues to the biogenesis of the lolitrem neurotoxins, and information on the structure-activity relationships within the indole-diterpenoids.     

Purification and characterization of an antifungal protein, C-FKBP, from Chinese cabbage

An antifungal protein was isolated from Chinese cabbage (Brassica campestris L. ssp. pekinensis) by buffer-soluble extraction and two chromatographic procedures. The results of matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry revealed that the isolated Chinese cabbage protein was identical to human FK506-binding protein (FKBP). A cDNA encoding FKBP was isolated from a Chinese cabbage leaf cDNA library and named C-FKBP. The open reading frame of the gene encoded a 154-amino acid polypeptide. The amino acid sequence of C-FKBP exhibits striking degrees of identity with the corresponding mouse (61%), human (60%), and yeast (56%) proteins. Genomic Southern blot analyses using the full-length C-FKBP cDNA probe revealed a multigene family in the Chinese cabbage genome. The C-FKBP mRNA was highly expressed in vegetative tissues. We also analyzed the antifungal and peptidyl-prolyl cis-trans isomerase activity of recombinant C-FKBP protein expressed in Escherichia coli. This protein inhibited pathogenic fungal strains, including Candida albicans, Botrytis cinerea, Rhizoctonia solani, and Trichoderma viride, whereas it exhibited no activity against E. coli and Staphylococcus aureus. These results suggest that recombinant C-FKBP is an excellent candidate as a lead compound for the development of antifungal agents.     

Structure elucidation and antifungal activity of an anthracycline antibiotic, daunomycin, isolated from Actinomadura roseola

The actinomycete strain Ao108 producing antifungal metabolites active against some plant pathogenic fungi was identified as Actinomadura roseola, based on the analyses of morphological and physiological characteristics. The antibiotic Da2B that showed a strong antifungal activity was isolated from the culture broth and mycelial mats of A. roseola strain Ao108 using various chromatographic procedures. On the basis of (1)H NMR, (13)C NMR, and 2-D NMR correlation data, the antibiotic Da2B was confirmed to have the structure of an anthracycline antibiotic, daunomycin. In vitro antimicrobial spectrum tests showed that the antibiotic Da2B had substantial inhibitory activity (10 microg mL(-)(1) of MICs) against mycelial growth of Phytophthora capsici and Rhizoctonia solani. The antibiotic also showed antiyeast activity against Saccharomyces cerevisiae, but the growth of Candida albicans was not affected. Antibacterial activity was found only against Gram-positive bacteria. In the further evaluation of in vivo efficacy, application of the antibiotic Da2B effectively inhibited the development of Phytophthora blight in pepper plants. However, the control efficacy of the antibiotic against Phytophthora infection was somewhat less than that of metalaxyl. The antibiotic Da2B did not show any phytotoxicity on pepper plants even at 500 microg mL(-)(1).     

Interactions between phosphorus availability and an AM fungus (Glomus intraradices) and their effects on soil microbial respiration, biomass and enzyme activities in a calcareous soil

The interactions between soil P availability and mycorrhizal fungi could potentially impact the activity of soil microorganisms and enzymes involved in nutrient turnover and cycling, and subsequent plant growth. However, much remains to be known of the possible interactions among phosphorus availability and mycorrhizal fungi in the rhizosphere of berseem clover (Trifolium alexandrinum L.) grown in calcareous soils deficient in available P. The primary purpose of this study was to look at the interaction between P availability and an arbuscular mycorrhizal (AM) fungus (Glomus intraradices) on the growth of berseem clover and on soil microbial activity associated with plant growth. Berseem clover was grown in P unfertilized soil (−P) and P fertilized soil (+P), inoculated (+M) and non-inoculated (−M) with the mycorrhizal fungus for 70 days under greenhouse conditions. We found an increased biomass production of shoot and root for AM fungus-inoculated berseem relative to uninoculated berseem grown at low P levels. AM fungus inoculation led to an improvement of P and N uptake. Soil respiration (SR) responded positively to P addition, but negatively to AM fungus inoculation, suggesting that P limitation may be responsible for stimulating effects on microbial activity by P fertilization. Results showed decreases in microbial respiration and biomass C in mycorrhizal treatments, implying that reduced availability of C may account for the suppressive effects of AM fungus inoculation on microbial activity. However, both AM fungus inoculation and P fertilization affected neither substrate-induced respiration (SIR) nor microbial metabolic quotients (qCO₂). So, both P and C availability may concurrently limit the microbial activity in these calcareous P-fixing soils. On the contrary, the activities of alkaline phosphatase (ALP) and acid phosphatase (ACP) enzymes responded negatively to P addition, but positively to AM fungus inoculation, indicating that AM fungus may only contribute to plant P nutrition without a significant contribution from the total microbial activity in the rhizosphere. Therefore, the contrasting effects of P and AM fungus on the soil microbial activity and biomass C and enzymes may have a positive or negative feedback to C dynamics and decomposition, and subsequently to nutrient cycling in these calcareous soils. In conclusion, soil microbial activity depended on the addition of P and/or the presence of AM fungus, which could affect either P or C availability.     

Polyacetylenes from the Apiaceae vegetables carrot, celery, fennel, parsley, and parsnip and their cytotoxic activities

A dichloromethane extract of root celery yielded falcarinol, falcarindiol, panaxydiol, and the new polyacetylene 8-O-methylfalcarindiol. The structure of the new compound was established by one- and two-dimensional (1D and 2D) NMR, mass spectrometry, and optical rotation data. Nonpolar extracts of roots and bulbs of carrots, celery, fennel, parsley, and parsnip were investigated for their content of polyacetylenes by high-performance liquid chromatography with diode array detection (HPLC-DAD). All five species contained polyacetylenes, although carrots and fennel only in minor amounts. Additionally, the cytotoxicity of the four polyacetylenes against five different cell lines was evaluated by the annexin V-PI assay. Falcarinol proved to be the most active compound with a pronounced toxicity against acute lymphoblastic leukemia cell line CEM-C7H2, with an IC(50) of 3.5 micromol/L. The possible chemopreventive impact of the presented findings is discussed briefly.     

Increased ecological and health risks associated with potentially toxic trace elements in agricultural soil adversely affected by gold (Au) mining activities

Journal of Soils and Sediments - Zaozigou gold mine is located in the Qinghai-Tibet Plateau, and it is one of the largest gold (Au) mines in China. Agricultural soil pollution of toxic trace...     

Anthogorgienes A-O, new guaiazulene-derived terpenoids from a Chinese gorgonian Anthogorgia species, and their antifouling and antibiotic activities

Fifteen new guaiazulene-based terpenoids designated anthogorgienes A-O (1-15) were isolated from a Chinese gorgonian Anthogorgia sp., together with eight known analogues (16-23). The structural patterns were classified into monomers, dimers, and trimers, which were supposed to be generated from a precursor guaiazulene and followed by side-chain and nucleus oxidation and oxidative rearrangement. The structures of new compounds were elucidated on the basis of extensive spectroscopic (IR, MS, and 1D and 2D NMR) data analysis. A possible biogenetic relationship of the isolated compounds was postulated. Some of the compounds showed potent antifouling activities against the larval settlement of barnacle Balanus amphitrite , whereas their antibiotic activities were also evaluated.     

Insecticidal, anti-juvenile hormone, and fungicidal activities of organic extracts from different Penicillium species and their isolated active components.

Organic extracts from mycelium and culture broth of 21 Penicillium isolates have been tested for insecticidal, insect anti-juvenile hormone (anti-JH), and antifungal activities. Culture broth extracts were the most active, mainly against insects; nearly 25% of them have shown high entomotoxicity (100% mortality at 100 microg/cm(2)). A strong in vivo anti-JH activity against Oncopeltus fasciatus Dallas was detected in the culture broth extracts from P. brevicompactum P79 and P88 isolates. The two new natural products isolated from P79, N-(2-methyl-3-oxodec-8-enoyl)-2-pyrroline (1) and 2-hept-5-enyl-3-methyl-4-oxo-6,7,8,8a-tetrahydro-4H-pyrrolo[2,1-b]-1, 3-oxazine (2), possessed anti-JH and insecticidal activity, respectively, against O. fasciatus. Synthesized natural compound 1 has shown an ED(50) of 0.7 microg/nymph when assayed on newly molted fourth-instar nymphs of O. fasciatus. Promising biological activities have also been detected in the synthetic precursors.     

Aflatoxin production in six peanut (Arachis hypogaea L.) genotypes infected with Aspergillus flavus and Aspergillus parasiticus, isolated from peanut production areas of Cordoba, Argentina

Aflatoxin contamination is one of the main factors affecting peanut seed quality. One of the strategies to decrease the risk of peanut aflatoxin contamination is the use of genotypes with resistance to Aspergillus infection. This laboratory study reports the resistance to Aspergillus infection and aflatoxin contamination of six peanut genotypes inoculated with 21 Aspergillus isolates obtained from the peanut production region of Cordoba, Argentina. The resistance was investigated in the seed coat and cotyledons of three resistant genotypes (J11, PI 337394, and PI 337409) and three breeding lines (Manfredi 68, Colorado Irradiado, and Florman INTA) developed at the Instituto Nacional de Tecnologia Agropecuaria (INTA), Manfredi Experimental Station, Cordoba, Argentina. Resistance to fungal colonization and aflatoxin contamination was found to be associated with seed coat integrity in the PI 337394, PI 337409, and J11 genotypes, whereas the INTA breeding lines such as Colorado Irradiado showed a moderate resistance and the Manfredi 68 and Florman INTA genotypes the least resistance. Furthermore, another type of resistance associated with cotyledons was found only in the PI 337394 genotype.     

Nitrogenase activity associated with leaf,root, and rhizosphere soils in an Indian tropical forest

Summary N₂ fixation (acetylene reduction assay) by phylloplane microorganisms was measured in dominant and co-dominant plant species growing in a tropical rain forest. No significant acetylene reduction was recorded with intact leaf samples. Azotobacter sp., Beijerinckia sp., Derxia sp., and Klebsiella pneumoniae were isolated as phylloplane N₂-fixing bacteria. Azospirillum lipoferum was only isolated from soil samples containing the roots of Poaceae. Nitrogenase activity was recorded in culture derived from the roots and rhizosphere soil samples, although low acetylene reduction activity indicates that these associations did not provide large amounts of N to the systems studied.     

Multivariate statistical analysis correlating 238U, 232Th,and 40K equivalent activities in soil to geochemical data from an agricultural area

Journal of Soils and Sediments - Multivariate statistical analysis was used for the correlation of 238U, 232Th, and 40K equivalent activities to geochemical data from soil samples collected at...     

Variation of enzyme activities, CO2 evolution and redox potential in an Eutric Histosol irrigated with wastewater and tap water

An Eutric Histosol soil was irrigated for 4 years with municipal wastewater to compare its characteristics with a soil under natural rainfall that had never received wastewater and a soil that was irrigated with normal tap water. Four years irrigation of the soil with wastewater caused significant (P<0.001) increase in dehydrogenase, urease, acid and alkaline phosphate activities and CO₂ evolution, and reduced the redox potential (P<0.05). The influence of treatments and plant cover on soil properties were significant (P<0.05) under both salix and grasses, except for few properties (redox potential and urease and alkaline phosphatase activities). It is suggested that, although different toxicants, e.g. heavy metals, may accumulate in wastewater-treated soils, enrichment of soil with organic substances and nutrients stimulated CO₂ evolution and enzyme activities in the irrigated soil.     

Microbial dynamics in an anaerobic soil slurry amended with glucose, and their dependence on geochemical processes

Under anoxic conditions, microbial activities interact closely with geochemical reactions and consequently affect soils, underlying aquifers, and the atmosphere. Recent studies have noted the relationships between microbial biodiversity and environmental conditions, but the dynamics of numerous coexisting microbial groups in connection with soil biogeochemical processes has never been investigated. In this work, we investigated the dynamics of anaerobic microbial populations using a new method combining PCR-SSCP and epifluorescent direct counts, and analysed these results in the light of biogeochemical changes. Batch incubations were performed over an 8-day period on a calcic cambisol (WRB) incubated anaerobically, either without amendment (treatment C) or after adding glucose (treatment +G). In treatment +G, the predominant microbial processes included (i) NO₃⁻ and NO₂⁻ reduction during the first 12 and 24 h of incubation respectively, (ii) fermentations during the first 6 days with non-symbiotic N₂ fixation between days 1 and 6, enabling bacterial growth during this period, and probably (iii) reduction of FeIII by H₂ oxidation throughout the incubation period. In treatment C, microbiological and geochemical measurements revealed no prominent microbial activities, and the PCR-SSCP method led to complex bacterial density profiles without prominent peaks. In treatment +G, 78 microbial groups were distinguished; these were divided into seven sets (A to G) according to their dynamics. Bacteria belonging to sets A, E and F grew during the period of intense fermentation and were probably able to fix N₂, as is the case with Clostridium butyricum (set A). Bacteria belonging to sets B, D, and G were probably able to reduce FeIII to FeII with concomitant oxidation of H₂ into H₃O⁺, but unable to fix N₂. Two microbial groups in these sets were closely related to Clostridium favosporum (set B) and the genus Bacillus (set B). Bacteria belonging to class C were probably only able to reduce N oxide(s). Lastly, we obtained two similar estimates of the gross increase in microbial biomass by taking into account either (i) the sum of gross increases for the 78 microbial groups, or (ii) the energy yield of catabolic reactions minus the energy requirement for N₂ fixation.     

Consumption of fruit, vegetables, sweets and soft drinks are associated with psychological dimensions of eating behaviour in parents and their 12-year-old children

OBJECTIVE: We investigated associations between consumption of fruits, vegetables, sweets and soft drinks and the psychological dimensions of eating in parents and their children. The role of the parent's characteristics for their children's food intake was also explored. DESIGN: Food intake patterns were assessed by self-reported consumption of the respective foods. Eating behaviour was measured by the Dutch Eating Behaviour Questionnaire and self-esteem by Harter self-perception scale. The participants were 1441 families (mother, father and their 12-year-old child), and additionally 354 mothers and thirty fathers. RESULTS: Among parents, reported intake of fruit and vegetables were associated with restrained eating, higher self-esteem, and higher education and age. Intake of sweets was related to more external and less restrained eating, and for mothers also emotional eating. Parent's intake of soft drink was foremost related to a younger age, and also weakly associated with psychological characteristics. The food intake of parents was more important for the children's food intake than any other characteristics. However, children's intake of sweets showed clear-cut positive associations with external eating. CONCLUSIONS: Psychological dimensions of eating behaviour are associated with patterns of food intake, in particular for consumption of sweets, and are most prominent in the parents. The children's food intake mirrored their parents' intake. Being sensitive to external food cues may increase unhealthy food consumption in our society, whereas more restrained eating may indicate proneness or intention to healthier food choices among parents. Emotional eating may imply a proneness to consume sweets for comfort, in particular among mothers.     

不同母质类型水稻土酶活性及其与理化性质的关系

将选自湖南双季稻区6种典型成土母质发育的水稻土置于长沙市郊同一生态条件下,通过7年定位池栽试验,研究了不同母质类型对土壤酶活性的影响以及酶活性与土壤理化性质之间的关系。结果表明：不同母质类型对土壤蔗糖酶活性的影响达到显著水平(P<0.05),对酸性磷酸酶、脲酶及过氧化氢酶活性的影响达到极显著水平(P<0.01)。主成分分析表明不同母质发育的水稻土酸性磷酸酶及脲酶作为综合评价土壤酶活性的指标要优于蔗糖酶及过氧化氢酶。6种不同母质水稻土土壤酶综合得分排序为：板页岩>河流沉积物>花岗岩>第四纪红土>紫色页岩>石灰岩。相关性分析表明,土壤蔗糖酶活性与有机质、全氮含量呈显著的正相关关系(P<0.05)。脲酶活性与硝态氮、铵态氮含量呈显著的正相关关系(P<0.05),与pH值呈显著的负相关关系(P<0.05)。酸性磷酸酶活性与速效磷含量呈显著正相关关系(P<0.05),与pH值呈显著负相关关系(P<0.05)。     

Trichoderma communities in soils from organic, sustainable, and conventional farms, and their relation with Southern blight of tomato

The objectives of this research were to investigate the relationship between propagule numbers and genetic diversity of Trichoderma species and Southern blight of tomato caused by soilborne plant pathogen Sclerotium rolfsii in soils with long-term organic, sustainable, and conventional farms. Dilution plating was use to quantify the propagule numbers of Trichoderma, denaturing gradient gel electrophoresis (DGGE) and DNA sequence analysis were used to identify Trichoderma species, and greenhouse assay were conducted for soil suppressiveness to Southern blight. The propagule numbers of Trichoderma tend to be higher in soils from conventional farms. There was no clear separation for the propagule numbers of Trichoderma based on different management systems using canonical correspondence analysis (CCA). However, there was general separation for total microbial communities based on organic and conventional management systems using CCA. That suggests that the difference of soil suppressiveness to disease from organic, sustainable, and conventional farms is due to the difference of the total microbial diversity but not directly due to the Trichoderma populations in each farming system. The propagule numbers of soil Trichoderma did not significantly correlate with the diseases suppressiveness, although individual species of Trichoderma harzianum was shown to be related to disease suppressiveness. Moreover, several Trichoderma species were found in the soil tested based on DGGE and DNA sequence analysis. Trichoderma hamatum, T. harzianum, Trichoderma virens, and Trichoderma erinaecem were the most abundant species in tested soil.