Bioactivities of caffeic acid methyl ester (methyl-(E)-3-(3,4-dihydroxyphenyl)prop-2-enoate): a hydroxycinnamic acid derivative from Solanum melongena L. fruits

Feeding deterrent and growth inhibitory effects of caffeic acid methyl ester (CME) isolated from the fruit extract of eggplant, Solanum melongena L. along with two known phenolic acids, caffeic acid (CA) and chlorogenic acid (CG) and azadirachtin (taken as active control), were evaluated against Spodoptera litura Fab. and Achaea janata L. (Lep., Noctuidae). The structure of CME was determined on the basis of extensive spectroscopic (UV, FTIR, ¹H, and ¹³C NMR) data analysis. CME has exhibited strong feeding deterrent activity against S. litura and A. janata larvae with ED₅₀ values of 52.02 and 41.02 μg/cm² leaf area, respectively. CA and CG were relatively less effective compounds when compared with CME. The results also revealed that the isolated compound CME caused marked larval growth inhibition in S. litura and A. janata larvae after 7 days of feeding on a treated diet. The effect on growth of larvae was concomitant with the reduced feeding and digestibility of ingested food. So the activities of digestive proteases within the midgut of larvae were examined using specific substrates to understand the digestive physiology in the larval guts. The bulk of the activity was associated with serine proteases comprising trypsin-, chymotrypsin-, and elastase-like enzymes, which decreased in diets containing CME, whereas, interestingly increased with CG. From the results, we conclude that CME isolated from S. melongena fruit extract shows feeding deterrent and larval development inhibitory activities that may prove useful in the management of S. litura and A. janata larvae.     

A new leaf disease on Populus euphratica in south‐western Iran

During the winter of 2012, a leaf spot disease was observed on Euphrates poplar (Populus euphratica) in the forest areas of Khuzestan province, south‐western Iran, causing significant damage in the Karun's riverside forests. Symptoms consisted of necrotic dark brown, circular to oval, 5‐ to 10‐mm spots on both surfaces of the leaves. A fungus having distinct dictyospores similar to those produced by Alternaria spp. was observed. The morphological characteristics, as well as the phylogenetic analysis of the internal transcribed spacer (ITS1‐5,8S‐ITS2) region, confirmed the identity of the strains belonging to the species Alternaria alternata. Pathogenicity tests were conducted on the alive leaves of P. euphratica on the young branches, as well as on the detached leaves in Petri dishes, through inoculation with spore suspension. Target spot symptoms similar to those observed in naturally infected leaves were developed on the inoculated leaves seven to 10 days after inoculation in both the inoculation procedures. A. alternata was consistently re‐isolated from the spots. Interestingly, similar symptoms were observed 7 days after detached leaf treatment with droplets of 15‐day‐old fungal culture filtrate, suggesting the production of pathotoxic compounds by the fungus. To our knowledge, this is the first report of A. alternata causing leaf spot on Euphrates poplar in Iran.     

Antifeedant activity and toxicity of leaf extracts from <Emphasis Type="Italic">Porteresia coarctata</Emphasis> Takeoka and their effects on the physiology of <Emphasis Type="Italic">Spodoptera litura</Emphasis> (F.)

Antifeeding and toxic effects of chemical extracts from Porteresia coarctata Takeoka on the insect pest Spodoptera litura (F.) were investigated in the laboratory. Hexane extracts of P. coarctata leaves were re-dissolved in dimethyl sulfoxide and applied in water to leaves of Ricinus communis L. (castor). A concentration of 1,000 and 2,000 ppm extract exhibited significant antifeeding and toxic effects to third instar larvae when compared to the same concentration of dimethyl sulfoxide in water as control. One day after feeding on extract-treated castor leaves, the mortality of S. litura larvae was 50%. Mortality increased to 95% after 11 days at a dosage of 2,000 ppm. Compared to the control, third instar larvae fed on castor leaves treated with P. coarctata leaf extract at different concentrations showed significant reduction in protein and DNA content in the fat body and midgut tissues. Results demonstrate the insecticidal and antifeedant properties of P. coarctata leaf extract against the generalist pest S. litura leading to biochemical changes in the insect body. Thus, P. coarctata contains phytochemicals, and the extract, in crude and pure form, has potential as a botanical insecticide in alternative control strategies against Lepidoptera pests.     

Activity of a novel compound produced by Aspergillus parasiticus in the presence of red flour beetle Tribolium castaneum against Pseudomonas aeruginosa and coleopteran insects

Aspergillus is a promising source of bioactive secondary metabolites. The addition of insect-derived material in the broth culture could trigger the biosynthesis of insecticidal and antimicrobial compounds by entomopathogenic fungus (EF). Insects inhabit diverse niches and interact with various bacteria, for this reason, when a fungus kills an insect should inhibit the insect’s gut bacteria to prevent the insect consumption by bacteria. According with this hypothesis, the EF is able to produce substances that inhibit bacteria growth or bacteria virulence strategies. In the present, investigation is demonstrated that the addition of Tribolium castaneum Herbst (Coleoptera: Tenebrionidae) components (2 % w/v) in a culture of saprophytic fungus Aspergillus parasiticus MOR 3 induces the production of a novel compound present inside mycelium 2-(4-bromophenyl)-2-oxoethyl benzoate, that is reported here for the first time as a natural product of A. parasiticus. In addition, increased significantly the fungal extracellular production of undecyl 4-fluorobenzoate. The mycelium extract (ME) at 250 μg per g of diet on adult population of T. castaneum produced an alteration of the feeding behavior of coleopteran insects (Repellency index = +30). The fraction derived from ME (Fr2) that contained the organobromine compound, 2-(4-bromophenyl)-2-oxoethyl benzoate had the highest inhibition of the Pseudomonas aeruginosa virulence factors like elastase enzyme (49 %) at 5 μg/ml and biofilm formation (43 %) at 100 μg/ml. The results suggest that the extract from mycelium of a non-aflatoxigenic A. parasiticus MOR 3 strain is a potential candidate as food coleopteran repellent as well as an anti-virulence strategy of P. aeruginosa.     

Effects of a bio-assay guided fraction from Polygonum cuspidatum root on the viability,acid production and glucosyltranferase of mutans streptococci

Polygonum cuspidatum root has been traditionally used for the treatment of dental diseases in Korea. The purpose of this study was to evaluate effects of P. cuspidatum root on the development of dental caries, especially its effects against bacterial viability and caries-inducing factors of Strptococcus mutans and Strptococcus sobrinus. Among methanol extract of P. cuspidatum root and its fraction tested, ethyl acetate fraction, composed of polydatin, resveratrol, anthraglycoside B, and emodin, showed inhibitory effects on glycolytic acid production and glucosyltransferase activity of S. mutans and S. sobrinus in addition to antibacterial activities.     

Monitoring the fate of a 30-year-old truffle orchard in Burgundy: from Tuber melanosporum to Tuber aestivum

Truffles, i.e. tree root-associated fungal fruiting bodies, clearly range among the world’s most exclusive delicacies. Despite the quite restricted natural geographic occurrence of one of the most renowned fungal species, namely Tuber melanosporum, the development of inoculation procedures in the late 1960s made it possible to enlarge its production area in different countries. This was achieved by planting orchards with host tree seedlings colonized by the fungus. In the present work, we investigate the behavior of one of the earliest T. melanosporum orchards planted in Burgundy (France) over a long-term scale (more than 30 years). A picture of the orchard evolution was obtained by recording truffle yields and fungal morphotypes over the seasons and relating them to host-tree development and climate data. The most relevant results include the time-delayed, but rather fast replacement of inoculated T. melanosporum by naturally occurring T. aestivum and the key role of climate in the inter-annual variability of truffle production.     

In vitro effects of Laccaria bicolor S238 N and Pseudomonas fluorescens strain BBc6 on rooting of de-rooted shoot hypocotyls of Norway spruce

The ectomycorrhizal fungus Laccaria bicolor S238 N and the bacterium Pseudomonas fluorescens BBc6 were used separately and in combination to induce in vitro rooting of de-rooted shoot hypocotyls of Norway spruce (Picea abies (L.) Karst.). When the culture medium was supplemented with tryptophan, a precursor of indole-3-acetic acid (IAA) synthesis, the presence of the ectomycorrhizal fungus increased the percentage of hypocotyls forming roots; furthermore, both the fungal and bacterial inoculations enhanced the number of roots formed per rooted hypocotyl. Similar results were obtained by adding exogenous IAA (5 and 10 &mgr;M) to the rooting medium. After the rooting phase, the fungal inoculation enhanced adventitious root elongation and branching as well as the aerial growth of the cuttings. Pseudomonas fluorescens BBc6 had no effect on these parameters. The production of IAA by pure cultures of L. bicolor S238 N and P. fluorescens BBc6 was estimated by immunochemical analysis using specific anti-IAA antibodies. Both L. bicolor S238 N and P. fluorescens BBc6 synthesized IAA in pure culture and synthesis was stimulated in the presence of tryptophan. Thus, the effect of the fungus in stimulating adventitious root formation and subsequent elongation and branching can be attributed, at least partially, to the synthesis of IAA by the fungus. The finding that P. fluorescens BBc6 had no effect on root elongation and branching although it produced IAA suggests that either IAA was not the only parameter involved in the stimulation of these processes by L. bicolor S238 N or the bacterium produced other compounds that counteracted the stimulatory effects of IAA on root elongation and branching.     

Chemical factors in Pinus strobus inhibitory to Armillaria ostoyae

Fresh sapwood of Pinus strobus inhibited the growth of Armillaria osotyae. Compounds extracted from sapwood with ethyl acetate inhibited the fungus in bioassays. Pinosylvin monothyl ether as tentatively identified by thin layer chromatography and ultraviolet spectra was isolated from the extract and found to inhibit the growth of A. ostoyae.     

Bacterial strains isolated from cedar wood improve the mycelial growth and morphology of white rot fungus <Emphasis Type="Italic">Phlebia brevispora</Emphasis> on agar and liquid medium

Growth enhancement of white rot fungus Phlebia brevispora TN3F by two bacterial strains TN3W-8 and TN3W-14 which showed high identity with genus Pseudomonas and Enterobacter, respectively, was investigated in liquid and solid media at different glucose concentrations. The white rot fungus P. brevispora TN3F and the bacterial strains were isolated from same white rot-decayed cedar wood from the Tano Forest Science Station at University of Miyazaki. Confrontational assay between the fungus and the bacterial strains revealed a fungal growth enhancement phenomenon, in the direction of the bacterial cells, even without direct contact. This enhancement increased with decrease of glucose concentration, 0.5% glucose offering comparatively the most mycelial growth. In potato extract (PE) medium at 0.5% glucose concentration, along with mycelial growth enhancement and morphological change of P. brevispora TN3F, growth and morphological enhancement of fungus TMIC33929, another strain of P. brevispora, was achieved by the bacterial strain TN3W-14, although TMIC33929 originated from a different environment. Outside few reports of a positive interaction between white rot fungi and other microorganisms, this is a first report of such positive interaction in liquid medium. Furthermore, this also is the first report of a single bacterium having potential to positively affect mycelia growth of different strains of a white rot fungus.     

两种杨树树皮内生真菌多样性及优势种群动态变化

为了解健杨94(转基因抗虫杨94)、三倍体毛白杨2个杨树品种干部树皮内生真菌区系及其优势种群的季节变化情况,本研究利用组织分离法从2个杨树品种996块组织中分离内生真菌1 175株,健杨94和三倍体毛白杨分别分离612、563株.利用形态特征和分子生物学方法鉴定为15个属、35个分类单元,包括担子菌1个分类单元,子囊菌34个分类单元.2个杨树品种内生真菌优势种群包括链格孢、葡萄座腔菌、镰孢属真菌、间座壳属真菌等,其中,仅有链格孢、葡萄座腔菌和桑砖红镰孢是两品种共有的优势种群种类,而且优势种群会随季节变化而变化.在两品种的内生真菌中,链格孢、葡萄座腔菌是最为常见的优势种群.     

Effect of Alternaria alternata on the coccinellid pest Henosepilachna vigintioctopunctata and its implications for biological pest management

Twenty-eight spotted potato ladybird beetle (Henosepilachna vigintioctopunctata) is a polyphagous pest that infests various crop plants of the family Solanceae. It extensively infests Withania somnifera, a high-value medicinal plant of this family. Both the beetle and W. somnifera leaf spot pathogen (Alternaria alternata) are individually found to be closely associated with the foliar damage and may rarely infest the same leaf simultaneously. Beetles fed on diseased leaves showed high pupal and larval mortality. Furthermore, the development of the larvae into pupa and from pupa to adult was also delayed, which in turn delayed the overall development of the insect. Adult emergence from pupae is also reduced in the insects fed on diseased leaves. Since the fungal pathogen is host specific to W. somnifera and does not infect the other solanaceous crops, therefore this fungal stain can be used in designing bio-control strategies for the other solanaceous plants frequently infested by the beetle.     

Interaction of root-knot nematode and mycorrhizal fungi on common beanPhaseolus vulgaris L.

The interactions of root-knot nematode and VAM fungus on common bean plants (Phaseolus vulgaris L.), and the sequence of nematode infection were studied in the greenhouse. The inoculation with VAM fungus caused a significant increase in plant height and fresh weight compared with non treated plants.Meloidogyne infection unsignificantly decreased plant height and dry weight. When fungus was inoculated at 15 and 30 days prior toM. incognita infection, a significant increase in fresh weight was observed. There were no significant differences in total nitrogen (mg/plant) between plants inoculated withM. incognita alone or those inoculated with bothM. incognita and VAM fungus at the same time or 15 days after the fungus inoculation. Plants preinoculated with VAM fungus 30 days prior to nematode infection had a significant increase in total nitrogen. The inoculation with VAM fungus caused a significant increase in phosphorus content. However, it was significantly decreased in plants inoculated with nematode alone and in plants inoculated with VAM fungus andM. incognita at the same time. Gall index and final nematode population were significantly increased when nematodes were treated at the same time with fungus, although there were significant decrease in nematode final population and gall index when the plants were treated with nematodes at 15 and 30 days after mycorrhizal infection. A decrease in percentage of fungal colonization was observed when nematodes were inoculated with fungus at the same time.     

Sesquiterpenes from Vladimiria souliei and their inhibitory effects on NO production

As a part of our ongoing search for plant-derived compounds that inhibit nitric oxide production, the methanol extract of the roots of Vladimiria souliei was found to show significant inhibitory effects on INF-γ-induced nitric oxide production in murine macrophage RAW264.7 cells. Bioactivity-guided isolation of the extract yielded two most active sesquiterpenes, including a new compound, named souliene A (1) and alismol (2). Their structures were elucidated by spectroscopic methods (IR, ESIMS, HRESIMS, 1D and 2D NMR). Two isolates showed promising inhibitory effects on INF-γ-induced nitric oxide production in murine macrophage RAW264.7 cells.     

Induced resistance of Norway spruce,variation of phenolic compounds and their effects on fungal pathogens

Three clones of Norway spruce (Picea abies) were studied for their response to mass‐inoculation with the blue‐stain fungus Ceratocystis polonica. The effect of different pretreatments (fungal inoculation and wounding) before mass‐inoculation was investigated for their possible role in an acquired resistance reaction. Pretreated trees showed enhanced resistance to the subsequent mass‐inoculation relative to control trees that received no pretreatment. Furthermore, the fungal colonization of inoculated trees was less than that of wounded trees. The phenolic content of the bark, analysed by RP‐HPLC, was compared in trees receiving different treatments. Trees inoculated with C. polonica had higher average concentration of (+)‐catechin, taxifolin and trans‐resveratrol than wounded trees. Both inoculated and wounded trees had higher average concentrations of these compounds than control trees. The effect of the phenolic extract of Norway spruce bark on the growth of the root rot fungus Heterobasidion annosum and the blue‐stain fungi C. polonica and Ophiostoma penicillatum were investigated in vitro. Heterobasidion annosum was not negatively affected, and the extracts had fungistatic effects on the blue‐stain fungi. The growth of O. penicillatum was more inhibited than the growth of the more aggressive C. polonica.     

美洲大蠊乙酸乙酯提取物的分离和抑菌活性分析

采用活性追踪法和柱层析法,以金黄色葡萄球菌、大肠杆菌等5种细菌为供试菌种,对美洲大蠊乙酸乙酯提取物进行分离和抑菌试验.研究发现:美洲大蠊乙酸乙酯提取物对供试的革兰氏阳性菌具有一定的抑制作用,其中,对枯草芽孢杆菌的抑菌活性最好;经硅胶柱层析初分离后,从乙酸乙酯提取物获得了9个具有抑菌活性的组分,其中,组分Fr7对5种供试菌均显示了较好的抑菌活性;组分Fr7及其相邻组分Fr8经硅胶和凝胶柱层析进一步分离纯化后,得到了7个具有抑菌活性的不饱和脂肪酸类化合物,均对藤黄微球菌表现出最好的抑菌活性.结果表明:美洲大蠊乙酸乙酯提取物的抑菌活性物质为不饱和脂肪酸类化合物,对供试的革兰氏阳性菌的抑菌活性较强.     

Effect ofMelia azedarach extract on larval development and reproduction parameters ofSpodoptera littoralis (Boisd.) andAgrotis ipsilon (Hufn.) (lep. noctuidae)

Methanolic extract ofMelia azedarach fruits enriched by washing with petrolether and ethyl acetate was used for laboratory treatments of two lepidopteran pests in Egypt. The experiments were carried out with concentrations of 10, 15, 25, 50, 100 and 1000 ppm in a diet and compared with control insects. In both insects food consumption, weight gain and conversion of ingested food (ECI) in body matter decreased with increasing extract amounts. The conversion of digested food (ECD) was lowered gradually by using higher concentrations ofMelia extract. Some antifeedant activity was observed in larvae ofS. littoralis andA. ipsilon. The percentage of mortality increased with application of higher concentrations ofMelia extract in both species. Starting from 3rd larval instar the larvae of both species reduced significantly their weight until pupation in 25 ppm and higher extract concentrations, while the larval period was prolonged. The pupal weight was significantly reduced at 15, 25 and 50 ppm. At higher concentrations the larvae failed to pupate. Duration of pupal period was affected only inA. ipsilon. All reproduction parameters, as period of oviposition, fecundity, fertility and longevity of males and females were affected using emerged adults from treated larvae with concentrations of 10, 15 and 25 ppmMelia extract. InS. littoralis no adult emerged from pupae originated from larvae treated with 50 ppm and higher amounts and no larva hatched from eggs laid by adults treated with 25 ppmMelia concentration as larvae. In both species the oviposition period was shortened at 15 and 25 ppm extract, the fecundity and fertility were drastically reduced, and the longevity of males and females was reduced. Cross sections of the midgut showed that the epithelial cells are destroyed in both pests. This can be one of the reasons for the observed effects.     

Truffle seedling production method has long-term consequences for tree growth and root colonization

Cultivation of Burgundy black truffles (Tuber aestivum syn. T. uncinatum) in Midwestern agroforestry has the potential to provide important income not only to landowners, but also to the hospitality industry. Economically viable harvest depends upon both successful seedling colonization by the truffle fungus and successful growth of the fungus along with the extending root system. We evaluated an established hybrid oak (Quercus bicolor × Q. robur) truffière 5 and 6 years after planting in May 2005, to evaluate the effects of three seedling production methods on tree growth and root colonization by T. aestivum. Oak seedlings produced using two variations on the RPM^© (Root Production Method) substrate grew significantly faster in height and diameter, but were significantly less well colonized by the truffle fungus compared with saplings from seedlings produced by the “Typical” method. We found that 0.7–19.8 % of root tips (mean 5.1 %) sampled from saplings grown from Typical seedlings were colonized by T. aestivum. By 2010, two distinct canopy forms had developed: ‘upright’ versus ‘spreading’. Although root colonization was not related to canopy form, future fruit body production may be influenced by the greater soil shading provided by the spreading canopy form. A comparison of autumn/winter air temperatures in Missouri, USA with European conditions suggests that fruit body production in Missouri will likely be greatest from mid-September through mid-December. Soil pH modification by application of crushed limestone prior to planting was effective in maintaining average pH at 7.16 through 2010, a level consistent with fruit body production.     

Additional diet of the parasitoids (Hymenoptera: Braconidae) and the parasitizing of the Elm Bark Beetle (Coleoptera: Scolytidae)

Supplementing the diet of Coeloides scolyticida, Dendrosoter protuberans and Ecphylus silesiacus (Hymenoptera: Braconidae) with Parker's diet and the nectars of melliferous plants: Daucus carota L., Sinapis alba L. and Stachys recta L. had a significant effect on longevity, reproduction, developmental rate, and parasitizing of Scolytus scolytus (Fab.), Scolytus multistriatus (Marsh.) and Scolytus pygmaeus (Fab.) (Coleoptera: Scolytidae). These values were considerably lower for wasps fed with the nectar of Sinapis arvensis L. and Trifolium repens L. and the lowest for those without dietary additives. Parker's diet had the most significant effect on the parasitoids and parasitizing of the bark beetles. Nectar of S. recta and S. alba proved most suitable for C. scolyticida and D. protuberans, while E. silesiacus using the nectar from D. carota flower significantly increased parasitizing of S. pygmaeus. T. repens flowers were not a suitable additive for the diet of parasitoids. The parasitoids without additives in their diet had shorter lives, shorter development, and were less effective in reducing host numbers. E. silesiacus wasps had the longest development, and C. scolyticida the shortest. Generally, the parasitoids of the second generation had a slightly prolonged development (1?–?2 days).     

Field resistance of Spodoptera litura (Lepidoptera: Noctuidae) to organophosphates, pyrethroids, carbamates and four newer chemistry insecticides in Hunan, China

The present studies were carried out to evaluate resistance in the populations of Spodoptera litura Fab. (Lepidoptera, Noctuidae) from five districts of Hunan Province in China to various insecticides from 2010 to 2012 using a standard leaf dip bioassay method. For organophosphates and pyrethroids, resistance ratios compared with a susceptible Lab-BJ strain were in the range of 14–229-fold for organophosphates and 12–227-fold for pyrethroids. Similarly, relative low levels of resistance to emamectin, indoxacarb, and chlorfenapyr were observed in all five populations. In contrast, the resistance to carbamates (thiodicarb or methomyl) was significantly higher than that of organophosphates, pyrethroids and newer chemistry insecticides. The pairwise correlation coefficients of LC₅₀ values indicated that the newer chemistry insecticides and old generation insecticides were not significant except abamectin, which was negatively significantly correlated with methomyl. A significant correlation was observed between thiodicarb, methomyl, and deltamethrin, whereas resistance to bifenthrin showed no correlations with resistance to other insecticides except deltamethrin. The results are discussed in relation to integrated pest management for S. litura with special reference to management of field evolved resistance to insecticides.     

In vitro TAXOL production,by Pestalotiopsis breviseta — A first report

Coelomycetous fungi were screened for the production of TAXOL. TAXOL production of Pestalotiopsis breviseta fungi is confirmed by Ultra Violet (UV) spectroscopic analysis, Infra Red (IR) analysis, high performance liquid chromatography (HPLC), nuclear magnetic resonance (NMR) and LC–MASS spectroscopy. TAXOL isolated from the P. breviseta fungus was identical with authentic TAXOL and produces 0.064mg/L (0.128% dry weight of fungal mat).