Antifungal activity of strawberry fruit volatile compounds against Colletotrichum acutatum

Eight volatile products characterizing strawberry aroma, which is generated from the oxidative degradation of linoleic and linolenic acids by a lipoxygenase (LOX) pathway, were examined because of their antifungal activity against Colletotrichum acutatum, one of the causal agents of strawberry anthracnose. In this study, the effects of aldehydes, alcohols, and esters on mycelial growth and conidia development were evaluated. (E)-Hex-2-enal was found to be the best inhibitor of mycelial growth [MID (minimum inhibitory doses)=33.65 microL L(-1)] and of spore germination (MID=6.76 microL L(-1)), while hexyl acetate was the least effective of all volatile compounds tested (MID=6441.89 microL L(-1) for mycelial growth and MID=1351.35 microL L(-1) for spore germination). Furthermore, the antifungal activity of (E)-hex-2-enal on susceptibility of strawberry fruits to C. acutatum was also confirmed. The presence of these molecules in jars containing strawberry fruits inoculated with a suspension of spores inhibited the fungus growth and prevented the appearance of symptoms. Moreover, a study of the effects of (E)-hex-2-enal on conidial cells of C. acutatum was evaluated by transmission electron microscopy. This volatile compound altered the structures of the cell wall and plasma membrane, causing disorganization and lysis of organelles and, eventually, cell death.     

电子束辐照对灰葡萄孢菌分生孢子萌发活性及致病力的影响

以灰葡萄孢病菌(Botrytis cinerea)分生孢子为研究对象,采用0.5、1.0、2.0和3.0kGy剂量电子 束辐照灰葡萄孢分生孢子,分别测定5℃和25℃培养条件下辐照对灰葡萄孢分生孢子的萌发活性及致病力的影响.结果表明:25℃和5℃培养条件下,2.0kGy处理分生孢子完全萌发时间比对照分别延迟了5和9d;2...     

GC-MS analysis of essential oils from some Greek aromatic plants and their fungitoxicity on Penicillium digitatum

The isolated essential oils from seven air-dried plant species were analyzed by gas chromatography-mass spectrometry (GC-MS). Thymus vulgaris (thyme), Origanum vulgare (oregano), and Origanumdictamus (dictamus) essential oils were found to be rich in phenolic compounds representing 65.8, 71.1, and 78.0% of the total oil, respectively. Origanum majorana (marjoram) oil was constituted of hydrocarbons (42.1%), alcohols (24.3%), and phenols (14.2%). The essential oil from Lavandula angustifolia Mill. (lavender) was characterized by the presence of alcohols (58.8%) and esters (32.7%). Ethers predominated in Rosmarinus officinalis (rosemary) and Salvia fruticosa (sage) essential oils, constituting 88.9 and 78.0%, respectively. The radial growth, conidial germination, and production of Penicillium digitatum were inhibited completely by oregano, thyme, dictamus, and marjoram essential oils at relatively low concentrations (250-400 microg/mL). Lavender, rosemary, and sage essential oils presented less inhibitory effect on the radial growth and conidial germination of P. digitatum. Conidial production of P. digitatum was not affected by the above oils at concentrations up to 1000 microg/mL. Apart from oregano oil, all essential oils were more effective in the inhibition of conidial germination than of radial growth. The monoterpene components, which participate in essential oils in different compositions, seem to have more than an additive effect in fungal inhibition.     

In vitro sensitivity of Botrytis cinerea to anthraquinone and anthrahydroquinone derivatives

The effect on mycelial growth of the fungus Botrytis cinerea of a set of structurally related tricyclic hydroquinones [9,10-dihydroxy-4,4-dimethyl-2,3,5,8-tetrahydroantracen-1(4H)-one and 9,10-dihydroxy-4,4-dimethyl-5,8-dihydroanthracen-1(4H)-one derivatives] and tricyclic quinones [4,4-dimethylanthracen-1,9,10(4H)-trione derivatives] was studied. In general, the anthraquinones presented higher activity than the anthrahydroquinones. Anthraquinone and anthrahydroquinone derivatives with methyl groups on the A ring showed higher antifungal activity than the unsubstituted ones, 4,4,6,7-tetramethyl-(4H)-anthracene-1,9,10-trione being the most active compound of this set. The presence of a polar group such as hydroxymethyl reduced the activity. The effect of two anthrahydroquinones and two anthraquinones on the conidia germination of the fungus was also determined. Anthrahydroquinones did not affect the germination. The most active compound was 4,4-dimethylanthracene-1,9,10(4H)-trione, with 100% inhibition of germination at 7 h of incubation. These results again suggest that the structure of the anthraquinones is important in exerting an antifungal effect on B. cinerea. Furthermore, possible mechanisms of action of compound 4,4-dimethylanthracene-1,9,10(4H)-trione were studied. This compound did not produce lipoperoxidation of membrane and did not induce the formation of oxygen reactive species, but it was able to permeabilize the plasmatic membrane of B. cinerea, increasing the phosphorus concentration in the intracellular medium.     

The response of cylindrocladium conidia to soil fungistasis

Direct observation of washed conidia of Cylindrocladium scoparium on non-sterile soils, air dried and rewetted immediately before deposition of conidia, indicated that peak germination (33–58%) occurred after 24 h incubation at 26°C. Peak germination on continually moist soils was lower (18–26%) than on rewetted soils. Lysis of germ tubes and germinating conidia on continually moist soils at 26°C was evident with 48 h. Conidia did not germinate on continually moist soils at 6°C and lysis did not become apparent until 168 h. Conidia germinated at a high level (93–99%) in axenic culture in the absence of exogenous C and N sources. The inhibition of conidial germination on soils may be attributed, in part, to the presence of soil volatiles. Germination of conidia placed on washed agar disks and exposed to volatiles from four soils ranged from 51 to 86% of the no-soil controls. Addition of carbon (13 ng C per conidium as glucose) and nitrogen (65 pg N ng^?1 C as NH₄C1) nullified the inhibitory effect of the soil volatiles. Germinability assayed on a selective medium at 26°C of conidia in artificially infested soils (approximately 10⁴ conidia g^?1 soil) decreased progressively during incubation at 26°C from 1 week to 4 months. No germinable conidia were recovered from artificially infested soils after 2 months incubation at 6°C. Conidia of C. floridanum and C. crotalariae responded similarly to C. scoparium in many assays.     

Influence of a volatile inhibitor in natural or limed soil on fungal spore and seed germination

Seed germination on sterilized moist filter or blotting papers resting on natural soils in stacked “Nalgene” plastic specimen dishes was inhibited particularly in alkaline soils in comparison with controls under similar conditions but without soil. The same response was observed when seeds were suspended on filter or blotting papers above the soils, suggesting the influence of an inhibitory volatile factor. Seed germination was not affected in Good's buffers over the pH range of the soil samples tested indicating that inhibition on filter or blotting papers in contact with soils was not due to the direct effect of pH. The observed inhibition of seed germination is similar to that found in the germination of conidia of common soil fungi; a volatile fungistatic factor was also detected in these soils. Liming of moderately acid soils increased conidial inhibition on indirect soil contact or from volatiles: germination of seeds, however, was not affected by liming in five of six soils tested. These results indicate that seeds of higher plants as well as fungal conidia are affected by some natural soils and volatiles are at least partially responsible for inhibition.     

Effect of 1-methylcyclopropene on volatile emission and aroma in cv. Anna apples

The rapidly ripening summer apple cultivar Anna was treated with 0.1 micro L(-1) and 1 microL L(-1) 1-methylcyclopropene (MCP) at harvest and kept at 20 degrees C, or stored for 5 weeks at 0 degrees C and then transferred to 20 degrees C. Total volatiles were not reduced by treatment with 0.1 microL L(-1) MCP, but were 70% lower in fruits treated with 1 microL L(-1) MCP than in untreated fruits. Ethylene production was 50% and 95% inhibited by 0.1 microL L(-1) and 1 microL L(-1) MCP, respectively. The volatiles produced by fruit at harvest were predominantly aldehydes and alcohols, with some acetate esters as well as 2-methyl butyl acetate and beta-damascenone. During ripening, the acetate and butyrate esters increased greatly and alcohols and aldehydes decreased. MCP-treated apples retained more alcohols, aldehydes, and beta-damascenone volatiles than did untreated apples. Sensory evaluation found that control and 0.1 microL L(-1) treated apples developed more fruity, ripe, and overall aromas, but the preference was for the 1 microL L(-1) treated apples with a less ripe aroma.     

Cinnamic acid inhibits growth but stimulates production of pathogenesis factors by in vitro cultures of Fusarium oxysporum f.sp. niveum

Long-term monoculture of watermelon leads to frequent occurrence of watermelon fusarium wilt caused by Fusarium oxysporum f.sp. niveum (FON). Some allelochemicals contained in watermelon root exudates and decaying residues are possibly responsible for promoting the wilt disease. The purpose of this study was to evaluate the allelopathic effect of artificially applied cinnamic acid on FON. Results demonstrated that hyphal growth of FON was strongly inhibited by cinnamic acid. At the highest concentration of cinnamic acid, the biomass in liquid culture was decreased by 63.3%, while colony diameter, conidial germination on plates, and conidial production in liquid culture were completely inhibited. However, mycotoxin production and activity of phytopathogenic enzymes were greatly stimulated. Mycotoxin yield, pectinase activity, proteinase activity, cellulase activity, and amylase activity were increased by 490, 590, 760, 2006, and 27.0%, respectively. It was concluded that cinnamic acid dramatically stimulated mycotoxin production and activities of hydrolytic enzymes by FON but inhibited growth and germination of FON. The findings presented here indicate that cinnamic acid is involved in promoting watermelon fusarium wilt.     

Interaction with and effects on the profile of proteins of Botrytis cinerea by C6 aldehydes

The natural volatile compounds cis-3-hexenal (c-3-H) and trans-2-hexenal (t-2-H) have significant antifungal activity with potential for use as postharvest fumigants of fruits and vegetables. However, the nature of their interaction with fungi and impact on fungal growth at the molecular level are largely unknown. The sites of interaction of these six carbon (C6) aldehydes with Botrytis cinerea, a common pathogen of many plant species, was characterized using 3H-labeled c-3-H and t-2-H. Radiolabeled C6 aldehydes were produced with lipoxygenase and hydroperoxide lyase extracts using [9,10,12,13,15,16-3H6]linolenic acid as a substrate. Following exposure of B. cinerea cultures to radiolabeled C6 aldehydes, radiolabel was recovered in protein-enriched but not lipid-enriched fractions. Radiolabel was incorporated at higher levels (6-fold per milligram of fresh weight and 4-fold per microgram of protein) into conidia than mycelia. About 95% of the radiolabeled aldehyde recovered in the protein fraction was from the surface of the fungal tissue, while 5% was from protein in internal tissue (cell wall, membrane, and cytosol). Exposure to t-2-H at both 5.4 and 85.6 micromol affected the protein profile of B. cinerea, changing the intensity of over one-third of all proteins. Both up-regulation and down-regulation of specific proteins were observed by two-dimensional gel electrophoresis, indicating a clear effect of t-2-H on changes in the protein profile of B. cinerea. This is the first evidence that fungal proteins are targets of the volatile C6 aldehydes and that sublethal levels of the aldehydes cause changes in the protein profile of a fungus.     

Structural changes in apple rings during convection air-drying with controlled temperature and humidity

The structure of heat pump dried apple slices, developed as a function of air temperature and constant humidity, was studied by measuring porosity and using electron microscopy. The porosity of the apple rings increased linearly when the moisture content decreased during drying and then reached a constant value. In all dried apple slices, a degree of cellular collapse occurred. Case hardening occurred in the surface of the dried tissue when the apple slices were dried at 40-45 and 60-65 degrees C, and in the extreme case (at 60-65 degrees C) cracks were formed on the surface.     

Induction of delta-cadinene synthase and sesquiterpenoid phytoalexins in cotton by Verticillium dahliae.

Phytoalexin biosynthesis occurred earlier in the resistant cotton cultivar Seabrook Sea Island 12B2 (SBSI) (Gossypium barbadense) than in the susceptible cotton cultivar Rowden (G. hirsutum) after inoculation with a defoliating isolate of the pathogen Verticillium dahliae. This was demonstrated by significantly higher levels of phytoalexins in SBSI 12 h after inoculation. Furthermore, by 48 h after inoculation of SBSI, the phytoalexins hemigossypol and desoxyhemigossypol achieved levels (23.9 and 10.5 microgram/g of fresh tissue, respectively) sufficient to completely inhibit conidial germination. Rowden required 96 h to attain comparable levels. Similarly, the activity of delta-cadinene synthase, a key enzyme required for the biosynthesis of the terpenoid phytoalexins, increased more rapidly in the resistant cotton cultivar than in the susceptible one. The changes in phytoalexin concentrations and enzyme activity are consistent with the hypothesis that phytoalexins are an essential component in protecting the plant from infection by V. dahliae.     

Investigations of aroma volatile biosynthesis under anoxic conditions and in different tissues of "Redchief Delicious" apple fruit (Malus domestica Borkh.)

Disks from different tissues were obtained from "Redchief Delicious" apple fruit (Malus domestica Borkh.) and analyzed for the ability to metabolize 1-pentanol as well as synthesize constitutive esters and alcohols under anoxic and aerobic conditions. The skin tissue displayed a greater capacity to synthesize pentanal, pentyl acetate, pentyl propionate, pentyl butyrate, and pentyl hexanoate than the hypanthial and carpellary tissues during incubation with 1-pentanol. With the exception of pentyl acetate and pentyl propionate biosynthesis, the hypanthial tissue synthesized these compounds at a higher rate than the carpellary tissue. Anoxia inhibited both constituent and 1-pentanol-derived ester biosynthesis. While anoxia inhibited ester biosynthesis, ethanol biosynthesis increased at a greater rate in tissue disks held under these conditions. Biosynthesis of 1-butanol, 2-methyl-1-butanol, and 1-hexanol was greater in tissue disks held in air during the first part of the measurement period and dropped off more rapidly than those transpiring in tissue disks held under anoxic conditions. The biosynthetic rates of all esters, both constituent and 1-pentanol-derived, increased as a result of air exposure. While hypoxic or anoxic conditions may promote ethanol synthesis, these conditions also appear to inhibit the formation of the ethanol-derived esters partially responsible for the off-flavor in apples attributed to ultralow O(2) controlled atmosphere storage.     

In vitro assessment of the inhibition of humic substances on the growth of two strains of <Emphasis Type="Italic">Fusarium oxysporum</Emphasis>

The effects of four humic substance (HS) samples, a soil humic acid and two humic acids and one fulvic acid isolated from a composting substrate, were evaluated on the mycelial growth of Fusarium oxysporum f. sp. melonis (FOM) and F. oxysporum f. sp. lycopersici (FOL). In general, any HS treatment reduced significantly the radial growth of the FOM mycelium either in normal [potato dextrose agar (PDA) medium] or sub-optimal (water–agar medium) nutritional conditions. Differently, the FOL growth, which was tested only on PDA, was either inhibited or stimulated on dependence of the HS treatment used. The HS fractions isolated from the composting substrate were the most effective inhibitors of mycelial growth of both fungi. Furthermore, any HS treatment was also able to alter the germination process of FOL in aqueous medium, not only by reducing significantly the number of viable germinating conidia but also by generally decreasing the rate of conidial germ-tube elongation. Apparently, the extent of the inhibitory action was related to some chemical and functional properties of HS, such as the COOH group content and elemental composition.     

Evidence for protein degradation by Botrytis cinerea and relationships with alteration of synthetic wine foaming properties

Botrytis cinerea is an important fungal pathogen particularly dreaded in the cool climate vineyard. It is responsible for important damage, especially the decrease in foamability of sparkling wines, such as Champagne. Different studies have shown that proteins are largely involved in the stabilization of Champagne foam despite their low concentration. Other works demonstrated changes in the electrophoretic characteristics of must proteins originating from botrytized grapes, although the cause of such alterations was never explained. In the first part of this study, results showed the release by B. cinerea of 3.5 mg/L total proteins in a synthetic liquid medium. Among these proteins, the presence of a protease activity on bovine serum albumin (BSA) and must proteins was demonstrated by using a colorimetric method and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. In the model wine, the Bradford method showed a BSA loss of 66% after 24 h and a loss of 96% after 120 h. In the same model wine, the soluble must protein concentration decreased by 35% after 1 week and by 53% after 2 weeks while the control showed no protein loss. B. cinerea proteases were then able to degrade BSA and must proteins and were above all active at must and wine pH and in the presence of ethanol and SO(2). The second part of this work was dedicated to the relationship between the presence of B. cinerea proteases and its effects on the synthetic wine foaming properties. The addition of a B. cinerea culture medium (1/33 v/v) to the synthetic wine containing 21 mg/L soluble grape proteins induced a decrease in foamability by 60% after 1 week. For BSA in the model wine, the foamability decreased by 32% after 24 h and by 95% after 120 h, as shown by the colorimetric method. These experiments demonstrate for the first time the relationship between B. cinerea protease activity and the decrease in wine foaming properties.     

Parallel synthesis: a new approach for developing analytical internal standards. Application to the analysis of patulin by gas chromatography-mass spectrometry

The polymer-assisted reaction of 4-(hydroxymethyl)furan-2(5H)-one (4HM2F) with 21 carboxylic acids using polystyrene-carbodiimide (PS-carbodiimide) yielded an ester library. Four of the esters, (5-oxo-2,5-dihydrofuran-3-yl)methyl acetate (IS-1), (5-oxo-2,5-dihydrofuran-3-yl)methyl butyrate (IS-2), (5-oxo-2,5-dihydrofuran-3-yl)methyl 2-methylpropanoate (IS-3), and (5-oxo-2,5-dihydrofuran-3-yl)methyl chloroacetate (IS-4), were tested as internal standards for the quantification of patulin in apple juice by gas chromatography-mass spectrometry in the selected ion monitoring mode (GC-MS-SIM). The developed method combines an AOAC official extractive step and a GC-MS-SIM analysis. Using a chromatographic column containing trifluoropropylmethylpolysiloxane as the stationary phase and IS-1 as the internal standard, it was possible to perform an accurate and precise quantification of underivatizated patulin in apple juice at concentrations down to 6 microg/L. A detection limit of 1 microg/L was established.     

Antifungal activity and fungal metabolism of steroidal glycosides of Easter lily (Lilium longiflorum Thunb.) by the plant pathogenic fungus, Botrytis cinerea

Botrytis cinerea Pers. Fr. is a plant pathogenic fungus and the causal organism of blossom blight of Easter lily (Lilium longiflorum Thunb.). Easter lily is a rich source of steroidal glycosides, compounds which may play a role in the plant-pathogen interaction of Easter lily. Five steroidal glycosides, including two steroidal glycoalkaloids and three furostanol saponins, were isolated from L. longiflorum and evaluated for fungal growth inhibition activity against B. cinerea, using an in vitro plate assay. All of the compounds showed fungal growth inhibition activity; however, the natural acetylation of C-6' of the terminal glucose in the steroidal glycoalkaloid, (22R,25R)-spirosol-5-en-3β-yl O-α-L-rhamnopyranosyl-(1→2)-[6-O-acetyl-β-D-glucopyranosyl-(1→4)]-β-D-glucopyranoside (2), increased antifungal activity by inhibiting the rate of metabolism of the compound by B. cinerea. Acetylation of the glycoalkaloid may be a plant defense response to the evolution of detoxifying mechanisms by the pathogen. The biotransformation of the steroidal glycoalkaloids by B. cinerea led to the isolation and characterization of several fungal metabolites. The fungal metabolites that were generated in the model system were also identified in Easter lily tissues infected with the fungus by LC-MS. In addition, a steroidal glycoalkaloid, (22R,25R)-spirosol-5-en-3β-yl O-α-L-rhamnopyranosyl-(1→2)-β-D-glucopyranoside (6), was identified as both a fungal metabolite of the steroidal glycoalkaloids and as a natural product in L. longiflorum for the first time.     

Polyphenolic antioxidants from the fruits of Chrysophyllum cainito L. (Star Apple)

Chrysophyllum cainito L. (Sapotaceae), known commonly as star apple or caimito, is a tropical tree that bears edible fruits. The fruits are grown commercially in certain tropical and subtropical areas, such as southern Florida. In this study, the fresh fruits were extracted with methanol and partitioned with hexane and ethyl acetate sequentially. The ethyl acetate soluble fraction displayed high antioxidant activity in the 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay (IC50 = 22 microg/mL). Activity-guided fractionation of the ethyl acetate soluble fraction was performed to identify the antioxidant constituents. Nine known polyphenolic antioxidants, (+)-catechin (1), (-)-epicatechin (2), (+)-gallocatechin (3), (-)-epigallocatechin (4), quercetin (5), quercitrin (6), isoquercitrin (7), myricitrin (8), and gallic acid, have been identified from the fruits. Of these nine antioxidants, 2 is present in the highest concentration in star apple fruits (7.3 mg/kg fresh weight), and 5 showed the highest antioxidant activity (IC50 = 40 microM) in the DPPH assay.     

超高压加工鲜切苹果片的色变动态特性

以红富士和冰糖心2种苹果为试材,研究0、200、300、400和500MPa超高压处理对鲜切苹果片的色变动态特性。结果表明,超高压处理能显著抑制鲜切苹果片在空气中的色变速率。采用4种三色值组合(ΔL*a*/b* 、ΔL*a*b*、Δ(ΔE)和ΔW.I.)作为色变指标研究鲜切苹果片的色变动态特性,结果表明,超高压处理和未处理的鲜切苹果片的各色变指标均随时间呈线性变化,鲜切苹果片的色变符合零级动力学反应。通过对三色值组合的优选,色变指标ΔL*a*/b*被选作反应压力对色变影响的最佳指标,并在此基础上建立了色变指标(ΔL*a*/b*)与压力(P)的关系模型。模型表明,400MPa为超高压加工鲜切水果片的较优压力。     

Responses of <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> f. sp. <Emphasis Type="Italic">niveum</Emphasis> to exogenously added sinapic acid in vitro

To assess the influence of phenolic acids from plant root exudates on soil pathogens, we studied the effect of sinapic acid added to chemically defined media on the growth and virulence factors of Fusarium oxysporum f. sp. niveum. Sinapic acid inhibited the growth and conidial formation and germination of F. oxysporum f. sp. niveum by 6.7–8.8% and 11.2–37.3%, respectively. Mycotoxin production by F. oxysporum f. sp. niveum was stimulated by 81.6–230.7%. Pectinase, proteinase, cellulase, and amylase activities were stimulated at a lower concentration of sinapic acid, while they were inhibited at a higher concentration. It is concluded that sinapic acid inhibited the growth and conidial germination of F. oxysporum f. sp. niveum and decreased the pathogenic enzymes’ activity at higher doses.     

Grape contribution to wine aroma: production of hexyl acetate, octyl acetate, and benzyl acetate during yeast fermentation is dependent upon precursors in the must

Wine is a complex consumer product produced predominately by the action of yeast upon grape juice musts. Model must systems have proven ideal for studies of the effects of fermentation conditions on the production of certain wine volatiles. To identify grape-derived precursors to acetate esters, model fermentation systems were developed by spiking precursors into model must at different concentrations. Solid-phase microextraction-gas chromatgraphy mass spectrometry analysis of the fermented wines showed that a variety of grape-derived aliphatic alcohols and aldehydes are precursors to acetate esters. The C6 compounds hexan-1-ol, hexenal, (E)-2-hexen-1-ol, and (E)-2-hexenal are all precursors to hexyl acetate, and octanol and benzyl alcohol are precursors to octyl acetate and benzyl acetate, respectively. In these cases, the postfermentation concentration of an acetate ester increased proportionally with the prefermentation concentration of the respective precursor in the model must. Determining viticultural or winemaking methods to alter the prefermentation concentration of precursor compounds or change the precursor-to-acetate ester ratio will have implications upon the final flavor and aroma of wines.