Cellular damage of plant pathogenic fungi by antifungal compounds produced by Bacillus spp. isolates

The use of microorganisms as biological control agents (BCAs) has become an effective alternative to chemical means of controlling plant pathogens. The antagonistic and inhibitory activity of 71 Bacillus spp. strains, which were isolated from different Mexican sites, were tested against several phytopathogen fungi, Fusarium oxysporum, Fusarium equiseti, Fusarium avenaceum, Bipolaris spp. and Alternaria spp. From the antagonism study, the strain ELI149 showed a marked inhibition of growth against all tested fungi; therefore crude metabolites from this strain were extracted using ethyl acetate and amberlite resin and probed against the same fungi as well as strains of Mucor sp., Penicillium spp. and Paecilomyces spp. The results indicated that amberlite was more suitable for extraction of secondary metabolites with antifungal activity. Finally, observation of cell damage in the tested pathogenic fungi showed marked morphological changes on reproductive structures in all tested fungi indicating that antibiosis was the mechanism of the antagonistic effect. These results suggest that metabolites from the Bacillus strains have a wide spectrum of antibiotic activities, which can be used as biocontrol agents for controlling fungal plant diseases of agricultural importance.     

The antifungal activity of the leaf gland volatile oil of sweet gale (Myrica gale) (Myricaceae)

Summary The antifungal activity of volatile oil obtained from the leaves of sweet gale (Myrica gale) has been investigated. Fungi, isolated from the leaves ofM. gale, were grown in liquid medium containing the oil and its effect on biomass production was gauged. The oil inhibited growth of all species of fungi to greater or lesser extent. Sesquiterpenes were more active than monoterpenes, and among the sesquiterpenes germacrone was marginally more active than -elemenone. These results support the proposition that the volatile oil ofM. gale has a role in resistance to fungal pathogens.     

Chemical composition and antifungal activity of Chelidonium majus extracts – antagonistic action of chelerythrine and sanguinarine against Botrytis cinerea

This study discusses the similarities and differences between the antifungal activity of Chelidonium majus L. extracts and chelerythrine/sanguinarine, the main alkaloids in these extracts, against the plant pathogenic fungus, Botrytis cinerea. The obtained data show that the Ch. majus L. extracts inhibit the growth of B. cinerea very effectively. The extracts activity is significantly greater than that of chelerythrine/sanguinarine standard solutions alone and it does not always reflect the total content of alkaloids in the extracts. It has been proved that B. cinerea exhibits very diverse resistance to chelerythrine and sanguinarine. The IC₅₀ values for chelerythrine and sanguinarine equal 5.41?mg/mL and 1.76?mg/mL, respectively. In the mixture the antifungal activity of chelerythrine and sanguinarine is antagonistic in the concentration dependent way. Based on the presented data it is not clear, however, which alkaloids present in the Ch, majus extracts at significantly lower concentration levels enhance the activity of chelerythrine and sanguinarine so that the extracts activity is considerably greater than that of both main alkaloids mixtures.     

Electrophysiological responses of four fungivorous coleoptera to volatiles of Trametes versicolor: implications for host selection

Fungi of the genus Trametes are known as important wood decomposers and are colonized by various species of Coleoptera and other arthropods. The aim of the present study was to investigate the importance of volatile chemical compounds as key attraction factors in recognition and host selection by species of Erotylidae (Dacne bipustulata, Tritoma bipustulata) as well as Cisidae (Sulcacis affinis) and Tenebrionidae (Diaperis boleti). Volatiles from freshly collected Trametes versicolor were collected by headspace sampling technique and identified by combined gas chromatography–mass spectrometry (GC–MS). To evaluate the biological significance of the volatiles we performed behavioural tests and recorded antennal responses of the fungus-inhabiting species by gas chromatography with electroantennographic detection (GC-EAD). The scent of T. versicolor was found to be dominated by sesquiterpenes; in GC-EAD 6 of these compounds elicited reproducible antennal signals in the tested species. Highly significant attraction effects to the fungus, the obtained odour samples and previously described fungal C₈-compounds were observed in behavioural tests. The possibility to detect these chemical compounds as a key cue for host selection implicate that beetles are able to discriminate between fungi of different age as well as different stages of colonization.     

Screening for plants and rhizospheral fungi with bioremediation potency of petroleum-polluted soils in a Tehran oil refinery area

The contamination of soils by toxic and/or hazardous organic pollutants, especially with crude oil, is a widespread problem. This study was conducted in a petroleum-contaminated area in a Tehran oil refinery to find petroleum-resistant plants and their rhizospheral fungal strains with bioremediation potency. The plants growing in the oil-polluted area were collected and determined taxonomically. Root samples of the plant species were collected from a polluted area and fungal strains determined by laboratory methods and taxonomical keys. The growth ability of the isolated fungal strains was studied in media containing 1%–15% crude oil. Results showed that seven plant species were of the highest density in the contaminated area: Alhagi persarum, Hordeum marinum, Peganum harmala, Phragmites australis, Prosopis farcta, Salsola kali, and Senecio glaucus. The root-associated fungi were isolated and showed that the fungal variation in the oil-polluted area is higher than that in a non-polluted area. The growth assay of isolated fungal strains showed that all studied fungal strains were able to form colonies at the applied concentrations but Alternaria sp. and Rhizopus sp. were the most resistant ones. Some plants were resistant to oil pollution, which also had positive effects on the fungal strains.     

Volatile organic compounds of Acacia longifolia and their effects on germination and early growth of species from invaded habitats

Acacia longifolia, a highly invasive species that invades coastal ecosystems in Mediterranean areas, produces significant impacts at different scales. Abundant foliage and thick canopies create a dense atmosphere that led us to hypothesise that the release of volatile organic compounds (VOCs) could play a role in the reduction of biodiversity observed in invaded areas. Therefore, we suggested that VOCs emitted by A. longifolia could exert inhibitory effects on physiological and biochemical parameters of native species. Using glass chamber bioassays, we evaluated the effect of aerial contact between VOCs from different plant parts of A. longifolia material and some native species. Volatile chemical composition was further analysed using GC-MS. Our results indicated that VOCs produced a notably reduction of seed germination. Furthermore, volatiles from leaves and flowers significantly decreased root length, shoot length and biomass for all species. Proline and malondialdehyde content did not significantly increase after contact with VOCs. Finally, chemical profile of VOCs from flowers, leaves and litter was significantly different, both qualitatively and quantitatively. As far as we know, our results constitute the first evidence of phytotoxicity induced by VOCs from A. longifolia, suggesting that flowers and leaves could influence its surrounding environment through VOCs release.     

In vitro degradation of the herbicide atrazine by soil and wood decay fungi controlled through Elisa technique

The interaction between atrazine, a triazine herbicide, and a series of decay fungi was characterized in terms of biodegradation of the herbicide and its influence on fungal growth. The following fungi were studied: thermophilic cellulolytic (Penicillium sp. 13) and noncellulolytic (Humicola lanuginosa sp. 5 and 12) strains isolated from self‐heated plant composts, mesophilic diphenol oxidase producing strain Mycelia sterilia INBI 2–26, white‐rot fungi Cerrena maxima, Coriolopsis fulvocinerea and Coriolus hirsutus. Competitive enzyme immunoassay was elaborated for detection of atrazine in cultural liquid. During agar plate cultivation the growth of Humicola sp. 5 was promoted by atrazine whereas the growth of Humicola sp. 12 and Penicillium sp. 13 was suppressed whereas M. sterilia INBI 2–26 was not affected by the herbicide. Neither atrazine‐accelerated nor atrazine‐depressed thermophilic strains decomposed atrazine during 21‐day cultivation according to ELISA data. In contrast, white‐rot fungi Coriolus hirsutus, Coriolopsis fuhocinerea and Cerrena maxima degraded nearly 50% of the herbicide in 5‐day submerged cultivation and 80–92% of the herbicide up to the 40th day. The soil strain M. sterilia INBI 2–26 decomposed 70% of atrazine in 17‐day cultivation. The degradation level depended of the time of atrazine introduction to the growing media. The relationships between the degree of atrazine decomposition and laccase and Mn‐peroxidase production were shown.     

Impact of deltamethrin on the endophytic fungal community of a Chinese cabbage,Brassica chinensis

Endophytic fungi, being closely associated with their host plants, are an important part of the ecosystem and food web. In modern agriculture, pesticides are widely used. The impact of pesticides on the endophytic fungal community of crops remains poorly understood. In this paper, Chinese cabbage Brassica chinensis, a popular leafy vegetable consumed worldwide, was selected for evaluating the potential effects of a pesticide, deltamethrin, on the endophytic fungal community. By culture method, 195 endophytic fungal strains classified into 39 taxa and 186 strains classified into 30 taxa, respectively, were isolated from the pesticide-treated and untreated B. chinensis samples. Sirodesmium spp. were the predominant endophytic fungi from both samples. The colonisation rates of endophytic fungi of the treated and the untreated samples were not significantly different, as determined by one-way analysis of variance (p?>?.05). The Shannon diversity indexes (H′) were close, being 2.216 and 2.152, respectively. However, the endophyte compositions of the treated and the untreated samples were significantly different (p?B. chinensis.     

Isolation and identification of fungi tolerant to polycyclic aromatic hydrocarbons and coal tar from different habitats in Lithuania

A group of 36 fungal strains, belonging to the Lithuanian mycobiota, was collected and isolated from different locations, habitats, and matrices, including creosote-treated wood in storage yards for crosstie wastes. The eight most perspective strains selected according to preliminary assessment of tolerance to coal tar were subsequently identified combining taxonomical evaluation and molecular techniques. The tolerance of the eight identified fungal species (five basidiomycetes and three ascomycetes) to the presence of various concentrations of coal tar, and for the four most perspective fungal strains (Pleurotus sp., Schizophyllum sp., Irpex lacteus, Bjerkandera adusta) to polycyclic aromatic hydrocarbons was evaluated. The ligninolytic enzymatic activity assay of the isolated strains resulted in a good correspondence between the tolerance to pollutants and the capability to produce ligninolytic enzymes indicating that this group of white-rot fungi is perspective for further investigation and eventual usage for mycoremediation of polycyclic aromatic hydrocarbons polluted substrates.     

Phytoremediation of petroleum-contaminated soils: Pre-screening for suitable plants and rhizospheral fungi

A field study was conducted in a petroleum-contaminated site in Kermanshah refinery (Iran) to find the petroleum-resistant plant species and the rhizospheral fungi for being used in bioremediation. Results showed that the amounts of petroleum pollution in non-vegetated areas is 6.8% and in vegetated areas is 2.2%. Plant samples were collected from petroleum-polluted areas and determined using morphological characters. It was found that eight plant species were growing on the contaminated sites: Polygonum aviculare, Amaranthus retroflexus, Noea mucronata, Alhaji cameleron, Crozophora heirosolymitrana, Poa sp., Convolvulus arvensis and Trifolium repense. The rhizospheral fungi of the plants were collected and determined using microscopic studies and taxonomical keys. The results determined the presence of 21 species in the rhizosphere of the plants growing in the polluted areas; three of these species were common in all of the plants and the others have species-specific distribution within the plants. The highest number of rhizospheral fungi (11 species) were determined for P. aviculare in non-polluted areas and nine species in polluted areas. It seems that the plant is the best candidate for using phytoremediation. However, the variation of fungi in petroleum-polluted areas was more than non-polluted zones. The culture of fungi in oil-contaminated media showed that, although all the studied fungi were resistant to low petroleum pollution (1% v/v), a few species, especially Fusarium species, showed higher resistance to petroleum pollution (10% v/v) and hence they may be suitable for mycoremediation in highly polluted areas.     

Indoor air quality in elementary schools of Lisbon in spring

Analysis of indoor air quality (IAQ) in schools usually reveals higher levels of pollutants than in outdoor environments. The aims of this study are to measure indoor and outdoor concentrations of NO₂, speciated volatile organic compounds (VOCs) and carbonyls at 14 elementary schools in Lisbon, Portugal. The investigation was carried out in May–June 2009. Three of the schools were selected to also measure comfort parameters, such as temperature and relative humidity, carbon dioxide (CO₂), carbon monoxide (CO), total VOCs, and bacterial and fungal colony-forming units per cubic metre. Indoor concentrations of CO₂ in the three main schools indicated inadequate classroom air exchange rates. The indoor/outdoor (I/O) NO₂ ratio ranged between 0.36 and 0.95. At the three main schools, the total bacterial and fungal colony-forming units (CFU) in both indoor and outdoor air were above the advised maximum value of 500 CFU/m³ defined by Portuguese legislation. The aromatic compounds benzene, toluene, ethylbenzene and xylenes, followed by ethers, alcohols and terpenes, were usually the most abundant classes of VOCs. In general, the indoor total VOC concentrations were markedly higher than those observed outdoors. At all locations, indoor aldehyde levels were higher than those observed outdoors, particularly for formaldehyde. The inadequate ventilation observed likely favours accumulation of pollutants with additional indoor sources.     

Synthesis and antimicrobial activity of esters of 3-ethoxy-4-hydroxybenzaldehyde oxime

A series of oxime esters was synthesized by esterification of 3-ethoxy-4-benzaldehyde oxime with acid chlorides in the presence of triethylamine. They were evaluated for their in vitro antifungal activity against three pathogenic fungi, namely Rhizoctonia bataticola, Fusarium udum, and Alternaria porii, and their antibacterial activity against the three bacterial strains Staphylococcus aureus, Pseudomonas aeruginosa, and Klebsiella pneumoneae. The compounds bearing medium-length alkyl chains showed greater activity than those with long alkyl chains. An electron-donating group in para position of the aryl ring confers highest activity.     

Floral scent variation in two <Emphasis Type="Italic">Antirrhinum majus</Emphasis> subspecies influences the choice of naïve bumblebees

Two wild subspecies of snapdragon, Antirrhinum majus, subspecies pseudomajus and striatum, differ in floral color and can be visually discriminated by insect visitors. The extent to which olfactory cues derived from floral scents contribute to discrimination between snapdragon subspecies is however unknown. We tested whether these two subspecies differ in floral scent and whether these olfactory differences are used by bumblebees (Bombus terrestris) to discriminate between them. We grew individuals of both subspecies, collected from a total of seven wild populations, under controlled conditions. We quantified the volatile organic compounds (VOCs) emitted by the flowers using gas-chromatography/mass-spectrometry/flame-ionization-detection. We studied antennal detection of VOCs by bumblebees, by means of electroantennogram study (EAG). We also performed behavioral experiments in a Y-maze to determine the innate response of bumblebees to the main floral VOCs emitted by our snapdragon subspecies. The floral scent of Antirrhinum majus pseudomajus contained three volatile benzenoids absent in the floral scent of Antirrhinum majus striatum. One of them, acetophenone, contributed over 69% of the absolute emissions of A. majus pseudomajus. These benzenoids elicited a significantly higher EAG response compared with other VOCs. In the Y-maze, bumblebees were significantly less attracted by acetophenone, suggesting an aversive effect of this VOC. Our findings indicate that bumblebees are able to discriminate between the two Antirrhinum majus subspecies. Differences in flower scent between these subspecies and olfactory bumblebee preferences are discussed in the light of biochemical constraints on VOCs synthesis and of the role of flower scent in the evolutionary ecology of A. majus.     

Electrophysiological responses and field attraction of the grey corn weevil, Tanymecus (Episomecus) dilaticollis Gyllenhal (Coleoptera: Curculionidae) to synthetic plant volatiles

The grey corn weevil, Tanymecus (Episomecus) dilaticollis, is an economically important polyphagous pest in Eastern and Central Europe. The present research is the first published electrophysiological study of olfactory sensitivity of adult T. dilaticollis to plant volatiles. The electrophysiological responses of male and female antennae were recorded to 20 synthetic volatile organic compounds (VOCs), previously identified to be released from the preferred host plants, maize and sunflower. Antennae of both sexes responded to a wide range of volatiles from different chemical classes: fatty acid derivatives, aromatic compounds and terpenoids. Two green leaf volatiles, (E)-2-hexen-1-ol and (Z)-3-hexen-1-ol, and one terpenoid, (±)-linalool, elicited the strongest electrophysiological responses by male and female antennae. Relatively high electrophysiological activity by T. dilaticollis antennae was also recorded to benzaldehyde, eugenol and caryophyllene. In the field, (E)-2-hexen-1-ol and (Z)-3-hexen-1-ol, when presented individually, caught significantly more male and female T. dilaticollis than unbaited controls. These compounds could be developed into an attractive trap for the detection and monitoring of T. dilaticollis.     

Source apportionment and ozone formation potential of volatile organic compounds in Lagos (Nigeria)

Concentrations of volatile organic compounds (VOCs) of the city air in Oba Akran road, Lagos were investigated. The chlorinated VOCs and xylene in Oba Akran were high. The average benzene/toluene ratio in Oba Akran was 1.7 suggesting that vehicular emission was a possible VOC source in most areas of Oba Akran. The toluene/(m+p-xylene) ratio also suggests a common source of toluene and xylene at these sites, presumably organic solvent used by industries located at Oba Akran. Our results showed that 7.10 tons of CO ₂ equivalents of VOCs are being emitted per year from Oba Akran. Xylenes were found to be the largest contributor to the ozone formation followed by trichloroethylene. A multivariate statistical analysis (Factor analysis extracted with Principal Component Analysis) has been applied to a set of data, and it was found that the main principal components, extracted from the air VOC pollution data, were related to gasoline and oil combustion/industrial activities.     

Removal of odors and VOCs in municipal solid waste comprehensive treatment plants using a novel three-stage integrated biofilter: Performance and bioaerosol emissions

• TSIBF was composed of ABRS, FRS and HBRS. • THIBF can effectively remove various odors, VOCs and bioaerosols. • Different reaction segments in TSIBF can remove different types of odors and VOCs. • TSIBF can reduce the emission of bioaerosols through enhanced interception. A novel three-stage integrated biofilter (TSIBF) composed of acidophilic bacteria reaction segment (ABRS), fungal reaction segment (FRS) and heterotrophic bacteria reaction segment (HBRS) was constructed for the treatment of odors and volatile organic compounds (VOCs)from municipal solid waste (MSW) comprehensive treatment plants. The performance, counts of predominant microorganisms, and bioaerosol emissions of a full-scale TSIBF system were studied. High and stable removal efficiencies of hydrogen sulfide, ammonia and VOCs could be achieved with the TSIBF system, and the emissions of culturable heterotrophic bacteria, fungi and acidophilic sulfur bacteria were relatively low. The removal efficiencies of different odors and VOCs, emissions of culturable microorganisms, and types of predominant microorganisms were different in the ABRS, FRS and HBRS due to the differences in reaction conditions and mass transfer in each segment. The emissions of bioaerosols from the TSIBF depended on the capture of microorganisms and their volatilization from the packing. The rational segmentation, filling of high-density packings and the accumulation of the predominant functional microorganisms in each segment enhanced the capture effect of the bioaerosols, thus reducing the emissions of microorganisms from the bioreactor.     

Wind tunnel attraction of grapevine moth females, Lobesia Botrana, to natural and artificial grape odour

Summary. Host plant volatiles which attract insect herbivores for egg-laying are of principal interest with respect to insect ecology and evolution. Direct applications concern population monitoring and control through behavioural manipulation. Identification of behaviourally active plant secondary metabolites is essential also for plant breeding for insect resistance. Grapevine moth females Lobesia botrana are attracted by upwind flight to green grape berries Vitis vinifera. The headspace of grape berries was collected on air filters. A solvent extract of these filters, released from a sprayer, attracted females in the wind tunnel. The results demonstrate that volatile cues mediate attraction of grapevine moth females to grape berries, and that headspace collections capture the essence of this odour signal. The air filter extracts were examined by gas chromatography coupled with electroantennographic detection, and the compounds eliciting a consistent antennal response in L. botrana females were identified by mass spectrometry. The headspace collection apparatus was calibrated for collection efficiency of the active compounds. Their recovery rate ranged from 35 % for methyl salicylate to 83 % for (E,E)-α-farnesene. A synthetic ten-component blend was then formulated. The blend consisted of compounds eliciting an antennal response, formulated in a blend ratio corrected for differences in collection efficiency. Subsequent wind tunnel tests showed that female attraction to this synthetic ten-component blend was not significantly different from attraction to grape berries, or to headspace collections of the same berries. At a release rate of 35 ng/h of the most abundant compound (E)-β-caryophyllene, 20 % of the test females approached the source of sprayed headspace collections and the ten-component synthetic blend, respectively. In comparison, 100 g of green berries, releasing the main compound (E)-β-caryophyllene at a rate of ca. 4.7 ng/h, attracted 10 % of the females by upwind flight followed by source contact.     

Synthesis and antifungal property of N,N′-bis(trans-cinnamaldehyde)-1,2-diiminoethane and its derivatives

N,N′-bis(trans-cinnamaldehyde)-1,2-diiminoethane and its derivatives were designed and synthesized. Fourier transform infrared spectroscopy (FTIR) and nuclear magnetic resonance (¹H-NMR) techniques were applied to characterize chemical structures, and the filter paper disk method was used to test for antifungal effect of the products against Aspergillus niger, Paecilomyces variotii, and Paecilomyces lilacinus. Chemical structure charaterization showed that the products are target compounds. Antimicrobial experiment results indicated that N,N′-bis(trans-cinnamaldehyde)-1,2-diiminoethane displayed reliable antifungal effects against three fungi, and its antifungal effect is close to that of cinnamaldehyde. N,N′-bis(p-methoxy cinnamaldehyde)-1,2-diiminoethane and N,N′-bis(p-chlorocinnamaldehyde)-1,2-diiminoethane also showed antifungal effects at a low concentration. Of the two, N,N′-bis(p-methoxy cinnamaldehyde)-1,2-diiminoethane appeared to be more suitable as it was more effective at higher concentrations against Paecilomyces.     

Response of non-enzymatic antioxidative mechanisms to stress caused by infection with Fusarium fungi and chemical protection in different wheat genotypes

The aim of this study was to assess the effect of infection with Fusarium fungi and chemical protection on the contents of phenolic acids and antioxidative potential of extracts from grains of 23 wheat genotypes. Based on chemical analyses including abiotic factors such as weather conditions and the location of the plantation, the authors attempted to clarify non-enzymatic antioxidative response mechanisms to two different stressors in 23 wheat genotypes. Based on chemical analysis, it was found that mass infection with fungi from the genus Fusarium induces defence mechanisms of the plant, which results in an increased production of phenolic acids, mainly those, which are substrates for the production of acids exhibiting antimicrobial action. This mechanism is confirmed by the analyses of antioxidant activity, which also increases significantly during fungal infection, connected with the production of mycotoxins. In the course of chemical plant protection, significant changes were observed in the production of phenolic acids. Even if antioxidant activity was not higher in comparison to the control, an increased production of certain phenolic acids was recorded, which suggests stimulation of the resistance system of the plant, at a simultaneous complete response of antioxidative mechanisms, as it takes place during fungal infection.     

Aggregation pheromone compounds of the black larder beetle Dermestes haemorrhoidalis Kuster (Coleoptera: Dermestidae)

Gas chromatography with simultaneous flame ionisation and electroantennographic detection (GC–EAD) and gas chromatography with mass spectrometry analysis (GC–MS) of abdominal extracts of adult male Dermestes haemorrhoidalis Kuster (Coleoptera: Dermestidae) revealed the presence of electrophysiologically and behaviourally active compounds to its conspecific males and females. Isopropyl dodecanoate (3), isopropyl (Z)-9-tetradecenoate (5), isopropyl tetradecanoate (6), isopropyl (Z)-9-hexadecenoate (7) and isopropyl hexadecanoate (8) were detected in male abdominal extracts only. Analysis of collected male headspace volatiles revealed the presence of six EAD-active compounds (3), (5), (6) and isopropyl tridecanoate (4) plus two unidentified compounds (1) and (9). Synthetic compounds (3), (4), (5), (6) and (7) showed EAD activity with antennae of both sexes in contrast to synthetic (8) which showed EAD activity with female antennae only. Male and female antennae of D. haemorrhoidalis reacted with high receptor potentials to isopropyl (Z)-9-dodecenoate (2), although this compound itself was detected in neither male nor female abdominal extracts or headspace volatiles. Petri dish bioassays indicated that male abdominal extracts and compounds (2), (3), (5) and (6) aroused and attracted conspecific male and female beetles significantly (P < 0.05) compared to female extracts. These results suggested the presence of a male-produced aggregation pheromone in D. haemorrhoidalis. Field assays with any of the described compounds, however, did not result in attraction of this beetle in significant numbers.