Antimycotic activities of Cinnamon-derived compounds against <Emphasis Type="Italic">Rhizoctonia solani</Emphasis> in vitro

In this study, the effects of medicinal plant extracts on the development of mycelium in the following phytopathogenic fungi were evaluated: Phytophthora capsici, Rhizoctonia solani, Fusarium solani, Colletotrichum gloeosprorioides, and Botrytis cinera. Of the 26 medicinal plants tested, six plant extracts showed antifungal activity against phytopathogenic fungi. The highest antifungal activity was exerted against R. solani by the n-hexane fraction of a Cinnamon (Cinnamomum cassia Blume) solvent extract. Therefore, the antifungal compound fractions I and II were purified from the n-hexane fraction by TLC on silica gel plates. When treated with solutions containing compound fractions I or II at a concentration of 2%, the mycelia growth rate of R. solani was reduced to 0.19 and 0.18, respectively. In addition, microscopic observation of the hyphal morphology of R. solani following treatment with compound fraction I revealed the presence of severely damaged hyphae. Specifically, the hyphal tips became swollen, collapsed or were completely destroyed in response to treatment with solution containing compound fraction I at concentration of 1%.     

Interaction Between Pratylenchus coffeae and Pythium aphanidermatum and/or Rhizoctonia solani on Chrysanthemum*

The interaction between Pratylenchus coffeae and Pythium aphanidermatum and/or Rhizoctonia solani on chrysanthemum and the influence of farmyard manure (FYM) on it was studied under pot conditions. The disease caused by P. aphanidermatum or R. solani was significantly high at varying inoculum levels of Pratylenchus coffeae. The severiy further increased when nematode infected plants were inoculated with both the fungi together. Organic manuring also influenced the disease severity which was greatest in soils containing 200% organic manure. The nematode population significantly increased in presence of R. solani, decreased in presence of P. aphanidermatum and remained unaffected when both the fungi occurred together around the plants. The nematode population was greatly reduced in soils containing 200% organic manure.     

<Emphasis Type="Italic">Exiguobacterium acetylicum</Emphasis> strain 1P (MTCC 8707) a novel bacterial antagonist from the North Western Indian Himalayas

Exiguobacterium acetylicum strain 1P (MTCC 8707) is a rhizospheric, Gram positive, rod shaped, yellow pigmented bacterium isolated from an apple orchard rhizospheric soil, on nutrient agar plates incubated at 4°C. The species level identification was arrived on the basis of 16S rRNA gene sequencing. The sequence showed 98% similarity with sequences of E. acetylicum available in the public domain. The strain was positive for siderophore and HCN production. In separate invitro assays it was found to inhibit the growth and development of Rhizoctonia solani, Sclerotium rolfsii, Pythium and Fusarium oxysporum. The volatile compound produced by the bacterium was found to be the most potent in inhibiting the hyphal development of R. solani, S. rolfsii, Pythium and F. oxysporum by 45.55, 41.38, 28.92 and 39.74% respectively. Commonly observed deformities caused by the diffusible and volatile compounds produced by the bacterium included hyphal inhibition, constriction and deformation. Under pot culture conditions the bacterium improved the germination and early growth parameters of pea (Pisum sativum) in the presence of R. solani and S. rolfsii.     

Pyrrolnitrin and phenazine production by Pseudomonas cepacia,strain 5.5B,a biocontrol agent of Rhizoctonia solani

Rhizoctonia stem rot of poinsettia caused by Rhizoctonia solani is controlled by strain 5.5B of Pseudomonas cepacia when poinsettia cuttings are rooted in polyfoam rooting cubes. Experiments were conducted to isolate and characterize secondary metabolites from strain 5.5B that were inhibitory towards R. solani. Inhibitory compounds were detected in fractions processed from liquid cultures of strain 5.5B. The most inhibitory compound isolated was pyrrolnitrin. A purple pigment consistently produced in culture by strain 5.5B was isolated and identified as 4,9-dihydroxyphenazine-1,6-dicarboxylic acid dimethyl ester, a phenazine. In vitro inhibition of R. solani occurred with the phenazine.     

Somatic Hybrids Between Potato and S. berthaultii Show Partial Resistance to Soil‐Borne Fungi and Potato Virus Y

Three tetraploid somatic hybrid lines produced by protoplast fusion between a dihaploid potato, Solanum tuberosum, cultivar BF15 and the wild potato species Solanum berthaultii were evaluated here for their response to different soil‐borne pathogens, that is Fusarium solani, Pythium aphanidermatum and Rhizoctonia solani as well as to infection by potato virus Y (PVY). Both hybrid and BF15 plants grown in vitro were inoculated with the tested pathogen strains, that is R. solani, P. aphanidermatum, or F. solani. The growth level and disease severity index of these plants were compared to the susceptible commercial cultivar Spunta. A better growth of inoculated hybrid plants and restricted disease symptoms were observed in comparison with the commercial plants. Under glasshouse conditions and after inoculation with R. solani and P. aphanidermatum, improved resistance of the hybrid plants to these pathogens was confirmed. Indeed, these plants showed no significant damage following inoculation and a better development in R. solani‐infected plants. The susceptibility of the hybrid tubers to R. solani, P. aphanidermatum, and to F. solani infection was also determined. A significant reduction of tissue colonisation was observed in all the hybrid lines compared to the cultivated cultivars. The STBc and STBd hybrids also showed improved resistance to the PVY ordinary strain (PVY^o) under glasshouse conditions.     

假单胞菌属双组分信号转导系统调控吩嗪生物合成研究进展

吩嗪是由假单胞菌或链霉菌产生的一类具有抗菌、抗肿瘤和抗寄生虫活性的含氮杂环代谢物,在农业和医疗领域具有广泛的应用.但吩嗪的合成受到复杂的级联网络调控.本文总结了假单胞菌属中双组分信号转导系统(two-component signal transduction system,TCS)对吩嗪生物合成的调控机制,阐明在双组分...     

A Wiskott–Aldrich syndrome protein is involved in endocytosis in Aspergillus nidulans

Endocytosis is vital for hyphal tip growth in filamentous fungi and is involved in the tip localization of various membrane proteins. To investigate the function of a Wiskott–Aldrich syndrome protein (WASP) in endocytosis of filamentous fungi, we identified a WASP ortholog-encoding gene, wspA, in Aspergillus nidulans and characterized it. The wspA product, WspA, localized to the tips of germ tubes during germination and actin rings in the subapical regions of mature hyphae. wspA is essential for the growth and functioned in the polarity establishment and maintenance during germination of conidia. We also investigated its function in endocytosis and revealed that endocytosis of SynA, a synaptobrevin ortholog that is known to be endocytosed at the subapical regions of hyphal tips in A. nidulans, did not occur when wspA expression was repressed. These results suggest that WspA plays roles in endocytosis at hyphal tips and polarity establishment during germination.     

Potential of Allyl Isothiocyanate to Control Rhizoctonia solani Seedling Damping Off and Seedling Blight in Transplant Production

The synthetic mustered flavouring essential oil, allyl isothiocyanate (AITC), was evaluated for its effect on suppression of Rhizoctonia solani growth in vitro, and in field soils for reducing inoculum density, saprophytic substrate colonization and seedling damping off and blight using snap bean and cabbage as indicator plants. In vitro growth was completely inhibited at the concentration of 50 μl/l. Inoculum density and saprophytic substrate colonization by the fungus in soil were not affected by AITC concentrations of 50 or 75 μl/kg soil. The inoculum density estimation by the use of soil‐drop technique created an artefact leading to an erroneous conclusion that the fungus was eradicated from soil within 1–3 days after AITC treatment at 150 or 200 μl/kg soil. The saprophytic substrate colonization showed that although the activity of R. solani was greatly reduced, the fungus still colonized 45% of the substrate units at these concentrations, and up to 100% at lower concentrations within 1 day after treatment. At higher concentrations the recovery rate from the substrates gradually declined over time to <6%. Drenching R. solani infested sandy‐loam or silty‐clay‐loam soil with water containing the emulsified AITC to provide 150 or 200 μl/l soil, a few days prior to planting, gave over 90% disease control in snap bean and cabbage, with no apparent phytotoxic effect. The effect of AITC was not influenced by the physical soil texture. AITC appears to have a good potential to replace methyl bromide fumigation of the substrate used for transplant production.     

Evaluation of antifungal potential of Marchantia polymorpha L., Dryopteris filix-mas (L.) Schott and Ephedra foliata Boiss. against phyto fungal pathogens

Methanol and flavonoid extracts (free and bound) of Marchantia polymorpha L., Dryopteris filix-mas (L.) Schott and Ephedra foliata Boiss. were screened against three fungal plant pathogens: Alternaria solani, Fusarium oxysporum and Rhizoctonia solani. The extracts from D. filix-mas and E. foliata showed >80% of mycelial inhibition of A. solani whereas M. polymorpha and D. filix-mas (rhizome) completely inhibited the mycelial growth of R. solani when tested at highest concentration (5 mg/ml). Inhibition of spore germination of fungi (A. solani and F. oxysporum) was observed to be 100% by most of the extracts at 10 mg/ml. Moreover, plant extracts were found effective in increasing seed germination and seed vigour simultaneously thereby decreasing the percentage of pathogen infection. The results of the present study reveal that the plants screened possess the potential to inhibit the crop fungal pathogens and further investigation is required to explore the biologically active constituents of these plants and to use them as natural plant protectants for agriculture.     

甲霜灵对大豆疫霉野生菌株及突变菌株生长发育影响的细胞学研究

本文利用电子显微镜技术研究了内吸性杀菌剂甲霜灵(Metalaxyl)对大豆疫霉Phytophthoras ojae野生菌株和突变菌株的形态学及超微结构的影响。结果表明:不同浓度甲霜灵处理后可导致野生菌株和突变菌株发生一系列不同的变化。低浓度(1μg/mL)处理后,野生菌株在培养基上的生长即可受到抑制,菌丝呈现不规则的肿胀、过度分枝;菌丝细胞壁不规则加厚,菌丝细胞内液泡增加,脂肪粒累积,细胞器排列紊乱,原生质最终坏死。随浓度的升高,野生菌株立即停止生长,菌丝干瘪坏死。而突变菌株只在高浓度(10μg/mL)甲霜灵处理后顶端菌丝出现少量较小的分枝,菌丝细胞壁无增厚现象,但细胞内脂肪粒大量积累,明显高于敏感性菌株;突变菌株在高浓度甲霜灵压力下仍继续生长。     

Inhibitory effect of essential oils of Eucalyptus sp. and Zataria multiflora on fungal growth of Macrophomina phaseolina

Macrophomina phaseolina is a causal agent for charcoal rot and cause economic damage in plants. In recent years, the great tendency for the use of biological substances to control of pests, weeds and the pathogens has been created. In this study, antifungal effects of concentrations 30, 60 and 120?μl/100?ml of essential oil of Eucalyptus sp. and Zataria multiflora were evaluated on the growth of M. phaseolina in vitro condition. The experiment was conducted on phactorial basis of completely randomised design with four replications. According to the results, a significant difference between inhibitory percentage of essential oils and different concentrations of essential oils on fungal colony growth has been observed (p?≤?0.01). Highest inhibitory percentage was related to the concentration of 120?μl/100?ml of Z. multiflora with 58.16% inhibition.     

Activity of Glucanases of Zygorrhynchus moelleri in Relation to Antagonism Against some Soil-Borne Plant Pathogenic Fungi

Zygorrhynchus moelleri antagonized some soil-borne fungi on agar and in composts. Filtrates of Z. moelleri cultures contained high levels of β-1,3-glucanase (exoglucanase) and β-1,3(4)-glucanase (endoglucanase) when the fungus was grown on hyphal wall material from Rhizoctonia solani or Pythium intermedium as sole carbon source. Lower exo- and endoglucanase activities were produced on Fusarium oxysporum f. sp. lini (F.o.l.) hyphal walls while insignificant levels were produced on Z. moelleri hyphal walls. Protease was also produced when Z. moelleri was grown on hyphal wall material, but chitinase or cellulase were not detected. Optimum production of both glucanases occurred in media at pH,6 to 7 and activity was optimal at pH 5. The enzymes were active over a temperature range in excess of 20–50 °C but optimum activity occurred at 40 °C. Cell free culture filtrates containing exo- and endoglucanase activity lysed living hyphal walls of R. solani, P. intermedium, Verticillium albo-atrum and F.o.l. although walls of the latter were least sensitive to lytic activity. The culture filtrates did not lyse living mycelium of Z. moelleri. Z. moelleri also produced substances inhibitory to growth of colonies of test fungi on agar. In glasshouse experiments disease severity in flax induced by R. solani and P. intermedium, in parsley by P. paroecandrum and in radish and Zinnia by P. intermedium was suppressed in composts inoculated with sporangiospore suspensions of Z. moelleri.     

Copper Oxide Nanoparticles Greener Synthesis Using Tea and its Antifungal Efficiency on Fusarium solani

Abstract

A great deal of research has been done on various uses of copper oxide. The synthesis of copper oxide nanoparticles was mediated using tea extract. The first sign of the reduction of copper ions to copper oxide was the change in color of extract to dark brown after treating with copper chloride. The resulting nanoparticles were characterized using X-ray diffraction (XRD) and Fourier transform infrared spectrometry (FTIR). Finally, the antimicrobial effects of these nanoparticles on Fusarium solani were studied in vitro by agar dilution method. The TEM images showed the synthesis of copper oxide with size of less than 80?nm. The synthesized copper oxide nanoparticles showed significant inhibitory effects on F. solani cultures so that the concentration of 80?μg/ml prevented approximately 90% of the mycelium growth of the fungus. The results showed that the inhibition zone of silver nanoparticles strongly depends on their concentration and increases by increasing the concentration of copper oxide nanoparticles in the medium.     

Antagonism against Rhizoctonia solani and fungitoxic metabolite production by some Penicillium isolates

A number of Penicillium isolates were recovered in association to Rhizoctonia solani strains pathogenic on tobacco and from soil on plates pre-colonized by the pathogen itself. Their antagonism toward R. solaniAG-2-1 was evaluated in dual cultures in vitro. Inhibition of growth was evident to some extent in most pairings, while hyphal interactions referable to mycoparasitic relationships were not observed. However, the occurrence of plasmolysis and/or vacuolisation and the induction of monilioid cells were indicative of the release of bioactive compounds. Therefore, production of fungitoxic metabolites was tested by adding concentrated culture filtrates of each Penicillium isolate to the growth medium of R. solani. Complete and lasting inhibition was incited by culture filtrates of some isolates belonging to P. brevicompactum, P. expansum, and P. pinophilum. Three purified compounds, respectively mycophenolic acid, patulin and 3-O-methylfunicone, which were extracted from culture filtrates, were able to inhibit R. solani in vitro. Their production was also detected in dual cultures of the same Penicilliumstrains with R. solani prepared in sterilized soil and when the Penicilliumstrains were cultured directly on R. solani mycelium harvested from liquid cultures. The possible role of such metabolites in antagonism of the above-mentioned Penicilliumspecies against R. solani is discussed.     

Enzymes Associated with Defence against Toxic Oxygen Species in PCNB-Tolerant and Sensitive Soil Fungi

The specific activities of enzymes such as superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPOX) and glutathione reductase (GR), which are involved in protection against toxic species of oxygen, were determined in mycelia extracts of pentachloronitrobenzene (PCNB)-tolerant and susceptible soil fungi. The organisms assayed were the highly PCNB-sensitive Rhizoctonia solani and Rhizopus arrhizus; Sclerotium rolfsii and Trichoderma harzianum, which are moderately susceptible to PCNB, and the fungicide-tolerant Fusarium oxysporum f. sp. melonis and Pythium aphanidermatum. No GPOX activity was detected in the six examined fungi. Significant differences in the specific activities of the other enzyme systems among the fungi were evident. Remarkably low levels of CAT activities were measured in R. solani. Except for T. harzianum, no meaningful differences regarding SOD, CAT and GR activities with age of the fungi cultures were observed. The electrophoretic patterns of SOD and CAT displayed dissimilarities among the fungi under study. P. aphanidermatum is more polymorphic with respect to both SOD and CAT enzyme systems as compared to the other fungi. The SOD of F. oxysporum f. sp. melonis, R. arrhizus and T. harzianum is a cuprozinc enzyme, while the mangano-SOD species was detected in S. rolfsii, R. solani and T. harzianum.     

Investigation of the infection process of macrophomina phaseolina on the surface of soybean roots using scanning electron microscopy

Sclerotial germination, hyphal growth pattern, and appressorial formation of the charcoal rot fungus, Macrophomina phaseolina (Tassi) Goid. on the surface of soybean roots, Glycine max (L.) Merrill was observed using scanning electron microscopy. Sclerotial germination is followed by a hyphal growth pattern which does not appear to be associated with any specific structural feature on the corrugated root surface. Appressoria are produced on the root surface at the tips of both primary hyphae and side branches within three days after inoculation.     

Apical assemblies of intermediate filament‐like protein FilP are highly dynamic and affect polar growth determinant DivIVA in Streptomyces venezuelae

Streptomyces spp. grow as branching hyphae, building the cell wall in restricted zones at hyphal tips. The organization of this mode of polar growth involves three coiled‐coil proteins: DivIVA and Scy, which form apical protein complexes referred to as polarisomes; and the intermediate filament‐like protein FilP, which influences cell shape and interacts with both Scy and DivIVA. Here, we use live cell imaging of Streptomyces venezuelae to clarify the subcellular localization and dynamics of FilP and its effect on hyphal morphology. By monitoring a FilP‐mCherry fusion protein, we show that FilP accumulates in gradient‐like zones behind the hyphal tips. The apical gradient pattern of FilP localization is dependent on hyphal tip extension and immediately dissipates upon growth arrest. Fluorescence recovery after photobleaching experiments show that FilP gradients are dynamic and subject to subunit exchange during vegetative growth. Further, the localization of FilP at hyphal tips is not directly dependent on scy, even though the strongly perturbed morphology of most scy mutant hyphae is associated with mislocalization of FilP. Finally, we find that filP has an effect on the size and position of the foci of key polar growth determinant DivIVA. This effect likely contributes to the phenotype of filP mutants.     

Resistance ofUrtica dioica to mycorrhizal colonization: a possible involvement ofUrtica dioica agglutinin

Roots of stinging nettle (Urtica dioica L.) were sampled at different sites around Basel (Switzerland) and examined under the microscope. They were completely devoid of mycorrhizal structures. Similarly, stinging nettle plants grown in the greenhouse in the presence of the arbuscular mycorrhizal fungusGlomus mosseae did not show any signs of mycorrhiza formation. Spread ofG. mosseae through the rhizosphere of stinging nettle plants was inhibited, and application of extracts of stinging nettle roots and rhizomes to hyphal tips ofG. mosseae reduced hyphal growth.Urtica dioica agglutinin, an antifungal protein present in the rhizomes of stinging nettle, inhibited hyphal growth in a similar way as the crude root extract. The possibility thatUrtica dioica agglutinin is at least partially responsible for the inability of stinging nettle to form the arbuscular mycorrhizal symbiosis withG. mosseae is discussed.     

Effects of the Plant Defence Inducer, Acibenzolar-S-Methyl, on Hypocotyl Rot of Soybean Caused by Rhizoctonia solani AG-4

The plant defence inducer, acibenzolar‐S‐methyl (ASM) was tested for its ability to protect soybean against hypocotyl rot caused by Rhizoctonia solani Kühn AG‐4. ASM in vitro exhibited an antifungal dose‐dependant activity in the form of reduced mycelial growth. This inhibition reached 40% in comparison with the control at a concentration of 0.5 g/l ASM in the growth media. Seed imbibition with ASM at a concentration of 0.08 g/l and 0.5 g/l significantly induced a reduction of the intensity of hypocotyl rot symptoms caused by R. solani AG‐4 that was correlated with a stimulation of chitinase activity. The protective effect of ASM against R. solani AG‐4 is probably due to the combination of induced resistance and its effect on pathogen growth. Seed treatment with ASM affected also the growth of 2‐day‐old seedlings. A dose‐dependant inhibition of the seminal root growth was observed which reached 53% at a concentration of 0.5 g/l ASM. This growth reduction of soybean was transitional and was rapidly recovered in optimal growth conditions except at 0.5 g/l of ASM.     

Antifungal activity of Bacillus thuringiensis strains and their efficacy against the cotton leaf worm Spodoptera littoralis

Bacillus thuringiensis strains that belong to B. thuringiensis, B. kurstaki and soil-isolated B.t. were assessed in the following phytopathogenic: Rhizoctonia solani, that had their mycelial growth decreased after incubation in the presence of the bacterial strains. The bacteria have also pathogenic effect against the insect pest Spodoptera littoralis. The isolate B.t. D-1 and the B.t. kurstaki HD-203 were found to be inhibiting R. solani, the strain B. kurstaki HD-203 displayed the highest percentage of inhibition (64%) and B.t. D-1 showed 49% of inhibition. Antagonistic activity was maintained up to pH 8.5, and the antifungal activity was stable to heat at 70?°C for 1?h. Minimal inhibitory concentrations were 152 and 131?μl/ ml for B.t. D-1 and B. kurstaki HD-203, respectively. The two strains also have high efficacy against S. littoralis larvae, B.t. D-1 gave 70% and the B. kurstaki HD-203 strain gave 80% mortality after seven days of treatment.