Evaluation of Eupatorium cannabinum Linn. oil in enhancement of shelf life of mango fruits from fungal rotting

Essential oils extracted from 17 higher plants belonging to different families were screened against Botryodiplodia theobromae and Colletotrichum gloeosporioides causing stem end rot disease and anthracnose disease in mango respectively. The essential oil of Eupatorium cannabinum was found to be fungitoxic in nature against both the mango-rotting fungi. Eupatorium oil was standardized through physico-chemical and fungitoxic properties. Gas Liquid Chromatography (GLC) analysis of the oil led to the identification of 16 components, which represented 77.97% of the oil. Germacrene D (16.11%) was found to be the major component. The oil showed a broad fungitoxic spectrum and was recorded to be more efficient than some synthetic fungicides. The oil also showed an inhibitory effect on pectinase and cellulase enzymes. The oil enhanced the shelf life of mango fruits by protecting from fungal rotting when tested as a fumigant. The LD₅₀ of Eupatorium oil was found to be 22.01 ml/kg body weight on mammalian mice.     

In Vitro toxicity of oil extracted from neem seeds collected from different locations across savanna agro-ecological zones of Nigeria on seed and soil-borne pathogens of cowpea (Vigna unguiculata L. Walp)

Graded extracts of neem seed oil extracted across the savanna agro-ecological zones (AEZs) of Nigeria were tested on mycelial growth of Colletotrichum capsici and Macrophomina phaseolina of cowpea in Nigeria. Minimum inhibition concentrations (MIC) and effectiveness levels of oil from AEZs were determined. Data were subject to ANOVA. Biplot was employed to access variation and interactions among the AEZs. Across the AEZs, fungitoxicity, MIC and effectiveness level varies; concentrations of 0.1 and 1.0% reduced the growth of C. capsici, while M. phaseolina was reduced at 10% concentration. On M. phaseolina, neem oil extracted from Ilorin, Ogbomosho (derived savanna) and Mokwa (southern guinea savanna) performed similarly, while a strong positive association existed between Ogbomosho (derived savanna) and Hadejia (Sahel savanna) samples. On C. capsici, samples from Bida (southern guinea savanna) and Ilorin (Derived savanna) were similar. This result could be a possible link between ecology, biodiversity and toxic principles in plant materials.     

Microbial conversion and in vitro and in vivo antifungal assessment of bioconverted docosahexaenoic acid (bDHA) used against agricultural plant pathogenic fungi

Microbial modification of polyunsaturated fatty acids can often lead to special changes in their structure and in biological potential. Therefore, the aim of this study was to develop potential antifungal agents through the microbial conversion of docosahexaenoic acid (DHA). Bioconverted oil extract of docosahexaenoic acid (bDHA), obtained from the microbial conversion of docosahexaenoic acid (DHA) by Pseudomonas aeruginosa PR3, was assessed for its in vitro and in vivo antifungal potential. Mycelial growth inhibition of test plant pathogens, such as Botrytis cinerea, Colletotrichum capsici, Fusarium oxysporum, Fusarium solani, Phytophthora capsici, Rhizoctonia solani and Sclerotinia sclerotiorum, was measured in vitro. bDHA (5 μl disc⁻¹) inhibited 55.30–65.90% fungal mycelium radial growth of all the tested plant pathogens. Minimum inhibitory concentrations (MICs) of bDHA against the tested plant pathogens were found in the range of 125–500 μg ml⁻¹. Also, bDHA had a strong detrimental effect on spore germination for all the tested plant pathogens. Further, three plant pathogenic fungi, namely C. capsici, F. oxysporum and P. capsici, were subjected to an in vivo antifungal screening. bDHA at higher concentrations revealed a promising antifungal effect in vivo as compared to the positive control oligochitosan. Furthermore, elaborative study of GC-MS analysis was conducted on bioconverted oil extract of DHA to identify the transformation products present in bDHA. The results of this study indicate that the oil extract of bDHA has potential value of industrial significance to control plant pathogenic fungi.     

γ-Asarone – the fungitoxic principle of the essential oil of Caesulia axillaris

The essential oil of Caesulia axillaris has exhibited its fungitoxicity against Aspergillus flavus at its minimum inhibitory concentration of 1300 mg/l. It showed the potentiality of an ideal fungitoxicant because of its long shelf life, thermostable nature, broad fungitoxic spectrum and persistence of fungitoxicity even on introduction of high inoculum density of the test fungus. The fungitoxic principle of the oil was standardized as -asarone which showed fungitoxicity against the test fungus at 500 mg/l.     

Selection of antagonistic soil microbes against a few fungal pathogens of piper betle L

Summary Against the fungal pathogens,Phytophthora parasitica, Sclerotium rolfsii, Colletotrichum capsici andGlomerella cingulata responsible for leaf and foot rot of betel vines, 3 fungus, 9 actinomycetes and 4 bacterial antagonists were screened out from 61 fungus, 28 actinomycetes and 4 bacterial organisms isolated from 3 sources of soils. One each from the 3 groups of antagonists when further tested, was found quite effective against the pathogens towards neutral side of H-ion concentration. The antagonists (P. citrinum, Streptomyces sp. and bacterial organism B-7) were also found effective against 18 among 32 isolated fungus organisms from potted soil (collected from betel vine soil) and except one which happened to beP. citrinum, the rest had no adverse effect upon their growth.This work is a part of a scheme supported by Food and Agriculture Council of Pakistan and conducted in Jute Research Institute.     

Evaluation of some essential oils as botanical fungitoxicants in management of post-harvest rotting of citrus fruits

During screening of some essential oils against Penicillium italicum, the oils of Mentha arvensis, Ocimum canum and Zingiber officinale were found to exhibit absolute fungitoxic activity against the test fungus. The oils were subsequently standardized through physico-chemical and fungitoxic properties. Practical applicability of the essential oils was observed in control of blue mould rot of oranges and lime fruits caused by P. italicum during storage. The Mentha oil-treated oranges and lime fruits showed enhancement of storage life of 6 and 8 days, respectively. The storage life of Ocimum oil-treated oranges and lime fruits was found to be enhanced by 6 days while in the case of Zingiber oil, it was 4 and 8 days enhancement of shelf life of oranges and lime fruits, respectively. This revised version was published online in August 2006 with corrections to the Cover Date.     

Biocontrol of collar rot disease of betelvine (Piper betle L.) caused by Sclerotium rolfsii by using rhizosphere-competent Pseudomonas fluorescens NBRI-N6 and P. fluorescens NBRI-N

Collar rot disease of betelvine (Piper betle L.) caused by Sclerotium rolfsii is difficult to control by conventional means by use of chemicals; therefore, use of biocontrol agents is desirable. In the present study, 186 bacterial strains of different morphological types were screened for their biocontrol activity against S. rolfsii under in vitro conditions. Two strains, Pseudomonas fluorescens NBRI-N6 and P. fluorescens NBRI-N, were selected for further studies because of their ability to inhibit the mycelial growth of the pathogen significantly. Spontaneous rifampicin-resistant (Rif) derivatives of P. fluorescens NBRI-N6 and P. fluorescens NBRI-N showing growth rate and membrane protein composition comparable to the wild type were selected to facilitate their monitoring in the rhizosphere. Field trials demonstrated that strain P. fluorescens NBRI-N6 was better than P. fluorescens NBRI-N in increasing the yield of betelvine significantly, whereas a consortium of the two strains controlled the disease more than either of the strains. The screening method should prove useful in identifying rhizosphere bacteria with the greatest potential for controlling diseases caused by phytopathogenic fungi.     

Fungitoxic and Phytotoxic Studies with Essential Oil of Ocimum adscendens

On screening the leaf extracts of some higher plants for their volatile antifungal activity against the test organism Aspergillus flavus, the extract of Ocimum adscendens exhibited the strongest fungitoxicity. The leaves showed maximum, fungitoxicity as compared with other plant parts. The volatile fungitoxic fraction obtained as an essential oil was standardized by various physico-chemical properties. The oil showed its fungicidal nature and broad range of activity at its minimum inhibitory concentration. The oil was thermostable and the toxicity remained unchanged even on autoclaving and on storage for up to 360 days. Moreover, the oil proved more active than some prevalent synthetic fungicides and exhibited no phytotoxic effect onVigna radiata.     

Fungitoxicity of some higher plants againstRhizoctonia solani

Summary Leaves ofChenopodium ambrosioides exhibited strong fungitoxicity against the mycelial growth ofRhizoctonia solani causing damping off diseases of some seedlings. Minimum inhibitory concentration of the fungitoxic constituent isolated in form of essential oil, was found to be 1000 ppm at which it was fungicidal in nature. It exhibited broad range of antifungal activity and did not show any phytotoxicity on the germination and seedling growth ofPhaseolus aureus.     

Individual and combined effects of berberine and santonin on spore germination of some fungi

Berberine and santonin were isolated from rhizomes ofBerberis aristata and unexpanded flower buds ofArtemisia maritima, respectively. Efficacy of these two chemicals individually as well as of their mixtures, was tested against spore germination of some saprophytic and obligate fungi. While berberine individually was effective against most of the fungi,Helminthosporium spp. were least affected even at the highest dose (1500 ppm). Santonin was equally effective against several fungi. Mixture of both alkaloids found to be more effective than individual ones. Keeping the dose of berberine constant and santonin at two different concentrations (viz. 250 and 500 ppm) the spore germination ofHelminthosporium, oryzae was stimulated. Increasing concentration of santonin inhibited the spore germination of all other fungi tested,Colletotrichum capsici being affected only by 20 and 5% (at berberine concentration of 250 and 500 ppm, respectively). On the other hand, santonin being constant and berberine at different concentrations, the mixture was effective against all the fungi.     

Effects of treatment with phenylthiosemicarbazide and 4'-chlorophenylthiosemicarbazide on Fusarium wilt of pea and tomato plants

Effects of treatment with phenylthiosemicarbazide (PTS) and its 4′-chloro-derivative (4′-chloro-PTS) on Fusarium wilt of pea and tomato plants were investigated. Depending on pH and availability of oxygen, PTS and 4′-chloro-PTS are converted to their corresponding phenylazothioformamides and phenylazothioformamide-S-oxides, which are the actual fungitoxic compounds. PTS and 4′-chloro-PTS were shown to inhibit growth of Fusarium oxysporum f. pisi and F. oxysporum f. lycopersici in liquid media as well as on agar plates at concentrations of 50–100 mg/1. Inhibition was greater at pH 7 than at pH 5. When administered to pea and tomato plants, both compounds caused severe phytotoxic effects, especially at temperatures favouring Fusarium wilt, thus almost entirely obscuring any protective activity against the diseases. All compounds were strongly adsorbed to loam, but readily released from sand. Neither in pea nor in tomato plants were PTS and 4′-chloro-PTS converted to any fungitoxic substance, not already present in the aqueous solutions administered.     

Schinus molle: a new source of natural fungitoxicant

The oil of Schinus molle exhibited the maximum fungitoxic activity during the screening of some essential oils against some common storage and animal pathogenic fungi. It showed absolute toxicity against animal pathogens and mild activity against storage fungi. The effective concentrations of the oil varied from 200 to 900 ppm. The toxicity of the oil persisted up to 80 degrees C and 90 days of storage but declined when autoclaved. It withstood heavy inoculum density. The oil exhibited a narrow range of activity and was found to be more effective than Multifungin, an antifungal drug. The oil was characterized by its various physicochemical properties. It was found to comprise 50 constituents. It appeared that some changes in the oil constituents during storage affected its fungitoxic potency.     

Schinus molle: a new source of natural fungitoxicant.

The oil of Schinus molle exhibited the maximum fungitoxic activity during the screening of some essential oils against some common storage and animal pathogenic fungi. It showed absolute toxicity against animal pathogens and mild activity against storage fungi. The effective concentrations of the oil varied from 200 to 900 ppm. The toxicity of the oil persisted up to 80 degrees C and 90 days of storage but declined when autoclaved. It withstood heavy inoculum density. The oil exhibited a narrow range of activity and was found to be more effective than Multifungin, an antifungal drug. The oil was characterized by its various physicochemical properties. It was found to comprise 50 constituents. It appeared that some changes in the oil constituents during storage affected its fungitoxic potency.     

Potential use of the essential oil ofTrachyspermum ammi against seed-borne fungi of Guar (Cyamopsis tetragonoloba L. (Taub.))

Essential oils isolated from leaves and seeds of seven umbelliferous plants were tested against the growth ofAspergillus flavus. Those from seeds ofTrachyspermum ammi, Cuminum cyminum, Carum carvi, Daucus carota and from leaves ofAnethum graveolens exhibited antifungal activity against the test fungus. Amongst these, oil from seeds ofTrachyspermum ammi was most toxic. Its minimum inhibitory concentration was 300 ppm, at which it exhibited fungistatic but not phytotoxic properties, when tested at 200, 300 and 400 ppm. The fungitoxic potency ofTrachyspermum seed oil remained unchanged after a long storage period and at high inoculum density of the test fungus. The oil was thermostable and was more efficaceous than the fungicides Agrosan G.N., Benlate, Ceresan, Dithane M-45 and Thiovit commonly used for the control of plant diseases.     

Zerumbone: a potential fungitoxic agent isolated from Zingiber cassumunar Roxb.

The rhizomes of Zingiber cassumunar exhibited strong fungitoxic action against Rhizoctonia solani, the damping-off pathogen. On chemical and spectral investigations, the antifungal compound was found to be zerumbone — a sesquiterpene. Its minimum effective dose against R. solani was 1000 ppm, much lower than some commercial fungicides. Zerumbone had fungistatic activity, a narrow fungitoxic spectrum and was not phytotoxic. Moreover, when used as a seed treatment, zerumbone could control damping-off disease of Phaseolus aureus caused by Rhizoctonia solani by 85.7%.     

Fungitoxicity of Some Higher Plants with Special Reference to the Synergistic Activity Amongst some Volatile Fungitoxicants

During screening of the leaves of 25 plant species for their volatile toxicity against the test pathogen Fusarium lateritium f.sp.cajani, Aegle marmelos (Ae), Citrus aurantifolia (Ci) and Mentha arvensis var. piperascens (Me) exhibited strong toxicity inhibiting the mycelial growth completely. The active volatile constituents from each plant were isolated in the form of essential oil and the fungitoxicity of each oil was tested separately. The Aegle oil was found fungistatic while Citrus and Mentha oils were fungicidal in nature. Three oil combination (1:1 v/v) viz., Ae-Ci, Ae-Me and Ci-Me were made and their fungitoxicity was tested. The oil combinations were found to be more fungitosic than the individual oils, which were fungistatic in nature. The Ci-Me combination exhibited a broad fungitoxic spectrum while the other two Ae-Ci and Ae-Me possessed a narrow range of toxicity. The oils were found to have no toxic effect on seed germination, seedling growth (root and shoot length), and general morphology of the host plant(Cajanus cajan).     

Fungitoxicity of Chemical Analogs with Heartwood Toxins

Trans-stilbene and tropolone as chemical analogs with naturally occurring fungitoxic heartwood compounds were studied with respect to their fungitoxic potency. While stilbene showed no fungitoxic activity towards the fungi Aureobasidium pullulans var. melanogenum, Penicillium glabrum, and Trichoderma harzianum in the concentrations tested, the minimal inhibiting concentration of tropolone was 10⁻³ M for Penicillium glabrum and Trichoderma harzianum, and 10⁻⁵ M for Aureobasidium pullulans var. melanogenum . In all cases, the effect of tropolone was a fungistatic one. Using chemical analogs for assessing the chemical basis of the fungitoxicity of tropolone, this substance proved to be the only compound tested which possesses fungitoxic properties. Received: 29 December 1997 / Accepted: 10 February 1998     

The volatile-producing Flavobacterium johnsoniae strain GSE09 shows biocontrol activity against Phytophthora capsici in pepper

Aims: Previously, we selected a bacterial strain (GSE09) antagonistic to Phytophthora capsici on pepper, which produced a volatile compound (2,4‐di‐tert‐butylphenol), inhibiting the pathogen. In this study, we identified strain GSE09 and characterized some of the biological traits of this strain in relation to its antagonistic properties against P. capsici. In addition, we examined bacterial colonization on the root surface or in rhizosphere soil and the effect of various concentrations of the volatile compound and strain GSE09 on pathogen development and radicle infection as well as radicle growth. Methods and Results: Strain GSE09 was identified as Flavobacterium johnsoniae, which forms biofilms and produces indolic compounds and biosurfactant but not hydrogen cyanide (HCN) with little or low levels of antifungal activity and swimming and swarming activities. Fl. johnsoniae GSE09 effectively colonized on pepper root, rhizosphere, and bulk (pot) soil, which reduced the pathogen colonization in the roots and disease severity in the plants. Various concentrations of 2,4‐di‐tert‐butylphenol or strain GSE09 inhibited pathogen development (mycelial growth, sporulation, and zoospore germination) in I‐plate (a plastic plate containing a center partition). In addition, germinated seeds treated with the compound (1–100 μg ml^?1) or the strain (10²–10¹⁰ cells ml^?1) significantly reduced radicle infection by P. capsici without radicle growth inhibition. Conclusions: These results indicate that colonization of pepper root and rhizosphere by the Fl. johnsoniae strain GSE09, which can form biofilms and produce indolic compounds, biosurfactant, and 2,4‐di‐tert‐butylphenol, might provide effective biocontrol activity against P. capsici. Significance and Impact of the Study: To our knowledge, this is the first study demonstrating that the Fl. johnsoniae strain GSE09, as a potential biocontrol agent, can effectively protect pepper plants against P. capsici infection by colonizing the roots.     

Baiting with Sclerotium rolfsii for selective isolation of Gliocladium virens from natural soil

A baiting technique for selective isolation of Gliocladium virens from natural soil was developed by mixing Sclerotium rolfsii —colonized sorghum grains with moist natural soil and incubating at 30 ± 5°C for 6–10 days. G. virens developed large, distinct colonies on the soil surface by colonizing S. rolfsii and was then easily isolated for use in biocontrol programmes. Trichoderma spp. were present in the soil but never developed conspicuous colonies on the soil surface, even when the soil was supplemented with large numbers of conidia.