Isolation of antifungal compounds from someZygophyllum species and their bioassay against two soil-borne plant pathogens

Roots and aerial shoots ofZygophyllum coccineum L.,Z. album L. andZ. dumosum Boiss were extracted with solvents of increasing polarity and tested against spore germination ofVerticellium albo-atrum andFusarium oxysporum f.sp.lycopersici. The effectiveness of each extract depends on the solvent, plant species, organ and test fungus. Petroleum ether extracts were ineffective as fungistatics while the methanol extracts possessed a high inhibitory effect toward spore germination suppression, depending on the fungus and plant species. Water or ethyl acetate root extracts ofZ. album caused a marked suppression of spore germination of the test fungi. Six saponins were isolated from the effective extracts on silica gel and Sephadex columns. They were identified by acid hydrolysis, alkaline saponification, IR and FAB MS. Quinovic acid was the dominant aglycone. Kaempferol was isolated from the flavonoid fraction ofZ. dumosum for the first time. Some of the isolated saponins and kaempferol provided successful control against the test plant pathogens under greenhouse experiment when used as seed treatment.     

Antifungal activity of alkyl gallates against plant pathogenic fungi

The antifungal activity of alkyl gallates against plant pathogenic fungi was evaluated. All of the fungi tested in this study were susceptible to some alkyl gallates, and the effect of linear alkyl gallates against plant pathogenic fungi was similar to the previously reported effects against Gram-negative and Gram-positive bacteria. We found that branched alkyl gallates showed stronger activity than did linear alkyl gallates with similar log P values. In addition, the antifungal activity of alkyl gallates was correlated with gallate-induced inhibition of the activity of mitochondrial complex II. The antifungal activity of alkyl gallates likely originates, at least in part, from their ability to inhibit the membrane respiratory chain.     

Antagonistic activity of Rhizobium spp. against beneficial and plant pathogenic fungi

Six different Rhizobium strains were tested for their antagonistic activities against 10 isolates of fungi. Their abilities to inhibit the growth of fungal isolates varied tremendously. Rhizobium legurninosnrum biovar phaseoli 6-3 showed the highest antagonistic activity which significantly reduced the mycelium dried weights of all the fungal isolates tested. This suggests that care should be taken when Rhizobium and fungal biocontrol agents are to be used together.     

Antimicrobial activity of a porcine myeloperoxidase against plant pathogenic bacteria and fungi

Porcine myeloperoxidase was evaluated for its antimicrobial activity against plant pathogenic bacteria and fungi. The results indicated that the enzyme, in the presence of a small amount of hydrogen peroxide, was effective against a broad spectrum of plant pathogens. The growth of seven bacterial species, including nine pathovars, from the genera Erwinia , Pseudomonas and Xanthomonas , was significantly inhibited by the enzyme at a concentration as low as 0·4 U ml⁻¹, while 4·0 U ml⁻¹ was lethal to all plant pathogenic bacteria examined. Myeloperoxidase, at 40 U ml⁻¹, was lethal to germinating spores from three isolates of the fungal plant pathogen Fusarium solani and two isolates from each of Colletotrichum gloeosporioides and C. malvarum . The enzyme's antifungal effects on the rice blast pathogen Magnaporthe grisea were studied both in vitro and on host plants. The enzyme significantly inhibited spore germination of two isolates of M. grisea races IC17 and IB49 at concentrations over 16 U ml⁻¹, and disintegration of fungal spore walls was caused by 80 U ml⁻¹. The enzyme was even more effective in reducing disease incidence of blast on young rice plants treated with 0·5 U ml⁻¹, while 2·5 U ml⁻¹ resulted in complete inhibition of infection. These results support and further extend the suggestion that myeloperoxidase could be used as a broad-spectrum biocontrol agent or as a transgenically expressed protein to combat diseases caused by plant pathogenic bacteria and fungi.     

Antifungal activity against plant pathogenic fungi of chaetoviridins isolated from Chaetomium globosum

The Chaetomium globosum strain F0142, which was isolated from barnyard grass, showed potent disease control efficacy against rice blast (Magnaporthe grisea) and wheat leaf rust (Puccinia recondita). Two antifungal substances were purified from broth from this organism and identified as chaetoviridins A and B. Chaetoviridin A exhibited higher antifungal activity than chaetoviridin B against plant pathogenic fungi both in vitro and in vivo. Treatment with chaetoviridin A at 62.5 microg/mL suppressed the development of rice blast and wheat leaf rust by over 80%. The molecule also exhibited moderate control of tomato late blight, resulting in 50% control following the application of 125 microg/mL chaetoviridin A.     

Antifungal activity of a novel compound from Burkholderia cepacia against plant pathogenic fungi

AIMS: To investigate antifungal activity of a novel compound (named as CF66I provisionally) against plant pathogenic fungi, mainly including Fusarium sp., Colletotrichum lindemuthianum, Rhizoctonia solani, etc. METHODS AND RESULTS: Minimal inhibition concentrations (MIC) and minimal fungicidal concentrations (MFC) of CF66I for each fungi were determined using serial broth dilution method. The data demonstrated MIC ranged from 2.5 to 20.0 microg ml(-1) and MFC were shown at levels of < or =7.5 microg ml(-1) except Fusarium sp. With reverse microscopy, profound morphological alterations of fungal cells were observed after exposure to CF66I. Conidiospores were completely inhibited, and protoplasm aggregated to form chalamydospores because of the changes of cell permeability. Some chalamydospores were broken, suggesting the compound probably possessed strong ability of damaging the cell wall. In addition, CF66I was investigated for its antifungal stability against Curvularia lunata. The results showed CF66I kept strong fungi-static activity over-wide pH range (pH 4-9) and temperature range (from -70 to 120 degrees C). CONCLUSIONS: The compound CF66I exhibited strong and stable broad-spectrum antifungal activity, and had a significant fungicidal effect on fungal cells. SIGNIFICANCE AND IMPACT OF THE STUDY: Results from prebiocontrol evaluations performed to date are probably useful in the search for alternative approaches to controlling serious plant pathogens.     

Isolation and characterization of Bacillus sp. producing broad-spectrum antibiotics against human and plant pathogenic fungi

A strain of bacterium producing antifungal antibiotic was isolated and identification of the strain was attempted. We could identify the bacterium as being a Bacillus sp., based on morphological observation, physiological characteristics, and 16S rDNA sequence analysis, thus leading us to designate the strain as Bacillus sp. AH-E-1. The strain showed potent antibiotic activity against phytopathogenic and human pathogenic fungi by inducing mycelial distortion and swelling and inhibiting spore germination. The antibiotic metabolite produced by the strain demonstrated excellent thermal and pH (2-11) stability, but was labile to autoclaving. From these results, we could find a broader antifungal activity of Bacillus genus. Isolation and characterization of the active agent produced by the strain are under progress.     

Effect of Mannich bases on some plant pathogenic fungi

3-(2-Pyridyl)-3-iminoisatin, 1-piperidinomethyl-3-(2-pyridyl)-3-iminoisatin, and 1-acetyl-3-(2-pyridyl)-3-iminoisatin affect spore germination ofAlternaria alternata, A. carthemi, Curvularia lunata, Fusarium oxysporum f.sp.cieri andF. udum and influence the development of powdery mildew (Erysiphe pisi) on pea under glasshouse condition as well as conidial germination ofE. pisi on excised pea leaves. Spore germination was inhibited in the sequence 1-acetyl-3-(2-pyridyl)-3-iminoisatin > 1-piperidinomethyl-3-(2-pyridyl)-3-iminoisatin > 3-(2-pyridyl)-3-iminoisatin followed the order accordingly. The powdery mildew development and conidial germination ofE. pisi 1-piperidinomethyl-3-(2-pyridyl)-3-iminoisatin > 1-acetyl-3-(2-pyridyl)-3-iminoisatin > 3-(2-pyridyl)-3-iminoisatin. The chemicals were compared with commonly used antifungal fungicides.     

Characterization and antimicrobial activity of water-soluble N-(4-carboxybutyroyl) chitosans against some plant pathogenic bacteria and fungi

Water-soluble N-(4-carboxybutyroyl) chitosan derivatives with different degrees of substitution (DS) were synthesized to enhance the antimicrobial activity of chitosan molecule against plant pathogens. Chitosan in a solution of 2% aqueous acetic acid-methanol (1:1, v/v) was reacted with 0.1, 0.3, 0.6 and 1 mol of glutaric anhydride to give N-(4-carboxybutyroyl) chitosans at DS of 0.10, 0.25, 0.48 and 0.53, respectively. The chemical structures and DS were characterized by ¹H and ¹³C NMR spectroscopy, which showed that the acylate reaction took place at the N-position of chitosan. The synthesized derivatives were more soluble than the native chitosan in water and in dilute aqueous acetic acid and sodium hydroxide solutions. The antimicrobial activity was in vitro investigated against the most economic plant pathogenic bacteria of Agrobacterium tumefaciens and Erwinia carotovora and fungi of Botrytis cinerea, Pythium debaryanum and Rhizoctonia solani. The antimicrobial activity of N-(4-carboxybutyroyl) chitosans was strengthened than the un-modified chitosan with the increase of the DS. A compound of DS 0.53 was the most active one with minimum inhibitory concentration (MIC) of 725 and 800 mg/L against E. carotovora and A. tumefaciens, respectively and also in mycelial growth inhibiation against B. cinerea (EC₅₀ = 899 mg/L), P. debaryanum (EC₅₀ = 467 mg/L) and R. solani (EC₅₀ = 1413 mg/L).     

硝酸咪康唑和克霉唑对临床常见分离真菌的体外抗真菌活性研究

硝酸咪康唑（Miconazole）和克霉唑（Clotrimazole）均属于咪唑类抗真菌药物,多年来广泛外用治疗皮肤癣菌病、皮肤和黏膜念珠菌病。为了解硝酸咪康唑和克霉唑对临床常见分离真菌的体外抗真菌活性,     

Mycosynthesis and characterization of silver nanoparticles and their activity against some human pathogenic bacteria

The aim of this study was to biosynthesis silver nanoparticles from the fungus Nigrospora sphaerica isolated from soil samples and to examine their activity against five human pathogenic strains of bacteria viz. Escherichia coli, Proteus mirabilis, Pseudomonas aeruginosa, Salmonella typhi and Staphylococcus aureus using disc diffusion method. The synergistic effect of silver nanoparticles in combination with commonly used antibiotic Gentamycin against the selected bacteria was also examined. The synthesized silver nanoparticles from free-cell filtrate were characterized by using UV–Vis spectrophotometer analysis, Fourier transform infrared spectroscopy (FTIR) and scanning electron microscope (SEM). UV–Vis spectrophotometer analysis showed a peak at 420 nm indicating the synthesis of silver nanoparticles, FTIR analysis verified the detection of protein capping of silver nanoparticles while SEM micrographs revealed that the silver nanoparticles are dispersed and aggregated and mostly having spherical shape within the size range between 20 and 70 nm. The synthesized silver nanoparticles exhibited a varied growth inhibition activity (15–26 mm diam inhibition zones) against the tested pathogenic bacteria. A remarkable increase of bacterial growth inhibition (26–34 mm diam) was detected when a combination of silver nanoparticles and Gentamycin was used. A significant increase in fold area of antibacterial activity was observed when AgNPs in combination with Gentamycin was applied. The synthesized silver nanoparticles produced by the fungus N. sphaerica is a promising to be used as safe drug in medical therapy due to their broad spectrum against pathogenic bacteria.     

Isolation and characterization of Bacillus sp. strain BC01 from soil displaying potent antagonistic activity against plant and fish pathogenic fungi and bacteria

Fungal and bacterial pathogens infect a diverse range of hosts including various plant and animal species. Fungal and bacterial diseases, especially of plants and aquatic animals, such as fish, lead to significant damage to crops and aquaculture, respectively, worldwide. The present study was conducted to isolate and characterize potent Bacillus strains with significant antagonistic activity against the major plant and fish pathogenic fungi and bacteria. We randomly collected 22 isolates of Bacillus from the soil, rhizosphere, and sediment from different parts of Bangladesh. Initial characterization, based on in vitro antagonistic activity on the culture plate, resulted in the selection of four gram-positive Bacillus sp. isolates. Among these, the isolate BC01, obtained from soil demonstrated the highest broad-spectrum anti-bacterial and anti-fungal activities. We confirmed the genus of BC01 to be Bacillus by morphological and biochemical tests as well as using molecular data analysis tools, including the study of 16s rDNA, phylogenetic relationship, and evolutionary divergence scores. The isolate significantly inhibited the mycelial growth of the plant pathogen, Penicillium digitatum and fish pathogen, Aphanomyces invadans in vitro. The anti-bacterial effect of the isolate was also evaluated against Pseudomonas spp. and Xanthomonas spp., the two deadliest plant pathogens, and Aeromonas veronii, Pseudomonas fluorescens, and Streptococcus iniae, three major fish pathogens that are primarily responsible for global aquaculture loss. The results of the present study could pave the way for developing potent drugs to combat microbial infection of plants and fish.     

In vitro activity of seven azole compounds against some clinical isolates of non-dermatophytic filamentous fungi and some dermatophytes

The in vitro activity of seven azole compounds viz clotrimozole, isoconazole, bifanazole, fluconazole oxyconazole, Bay n 7133 and Bay L 9139 was investigated against 47 clinical isolates of pathogenic non-dermatophytic filamentous fungi and dermatophytic fungi. The isolates included Hendersonula toruloidea-26, Scytalidium hyalinium-5, Scytalidium japonicum-1, Trichophyton rubrum-5, Trichophyton tonsurans-3, Trichophyton mentagrophytes var. mentagrophytes-4, Epidemophyton floccosum-2, Microporum gypseum-2 isolates. The drugs were significantly more active against the dermatophytes (MIC range 0.025–1.56 g/ml) than non-dermatophytes (MIC range 0.39–6.25 g/ml). Isoconazole showed more activity than the rest of the azole compound tested. Clotrimazole, fluconazole, oxyconazole, bifonazole were comparable in their inhibitory activity against both dermatophytes and non-dermatophytes. The azole derivatives, Bay n 7133 and Bay L 9139 showed higher MIC range i.e. gave a range of 0.39–1.56 g/ml for dermatophytes and 1.56–6.25 g/ml for non-dermatophytic filamentous fungi. The minimal fungicidal concentration (MFC) of all the drugs tested were mostly within 2–8 times their MIC values.     

Biosynthesis of silver nanoparticles by the endophytic fungus Epicoccum nigrum and their activity against pathogenic fungi

There is an enormous interest in developing safe, cost-effective and environmentally friendly technologies for nano-materials synthesis. In the present study, extracellular biosynthesis of silver nanoparticles was achieved by Epicoccum nigrum, an endophytic fungus isolated from the cambium of Phellodendron amurense. The reduction of the silver ions was monitored by UV–visible spectrophotometry, and the characterization of the Ag NPs was carried out by X-ray diffraction and transmission electron microscopy. The synthesized Ag NPs were exceptionally stable. It was found that an alkaline pH favored the formation of Ag NPs and elevated temperature accelerated the reduction process. Furthermore, the antifungal activity of the Ag NPs was assessed using a microdilution method. The biosynthesized Ag NPs showed considerable activity against the pathogenic fungi. The current research opens a new path for the green synthesis of Ag NPs and the process is easy to scale up for biomedical applications.     

链霉菌KIB-H1556次级代谢产物抗植物真菌活性研究

对曼陀罗(Datura stramonium L.)根际链霉菌Streptomyces sp.KIB-H1556的次级代谢产物进行研究,利用硅胶柱色谱、凝胶柱色谱和半制备HPLC等分离手段对其发酵产物进行分离纯化,采用MS和NMR等波谱学手段并结合文献数据鉴定了3个单体化合物的结构,分别为:Bafilomycin D(1)、Bafilomycin B1(2)和Bafilomycin B2(3)。初步抗植物病原真菌活性筛选发现化合物2和3具有广谱抗真菌活性,尤其对玉米病原真菌的抑制活性显著,可作为玉米病原真菌病害的潜在生物防治剂。     

Evaluation of antifungal activity of food additives against soilborne phytopathogenic fungi

The efficacy of eight food additives as possible alternatives to synthetic fungicides for the control of soilborne pathogens, Fusarium oxysporum f. sp. melonis, Macrophomina phaseolina, Rhizoctonia solani, and Sclerotinia sclerotiorum was evaluated in this study. A preliminary selection of food additives was performed through in vitro tests. The ED₅₀, minimum inhibition concentration (MIC), and minimum fungicidal concentration (MFC) values showed that ammonium bicarbonate and potassium sorbate were more toxic to soilborne pathogens compared to other food additives with few exceptions and, therefore selected for further testing in soil. The inhibitory and fungistatic efficacy potassium sorbate were higher than that of ammonium bicarbonate in in vitro tests. Potassium sorbate completely inhibited F. oxysporum f. sp. melonis, M. phaseolina, and R. solani at 0.6% in soil tests. In contrast ammonium bicarbonate at 0.6% was inferior compared to potassium sorbate. Ammonium bicarbonate achieved to control all fungi at 2% that is the highest concentration used in this study. Potassium sorbate showed higher toxicity to all fungi compared to ammonium bicarbonate in soil tests. Both ammonium bicarbonate and potassium sorbate increased the pH of soil. The rate of pH increase was higher in ammonium bicarbonate.     

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.     

Rhizobia as a biological control agent against soil borne plant pathogenic fungi

Rhizobia promote the growth of plants either directly through N2 fixation, supply of nutrients, synthesis of phytohormones and solubilization of minerals, or indirectly as a biocontrol agent by inhibiting the growth of pathogens. The biocontrol effect of rhizobia is due to the secretion of secondary metabolites such as antibiotics and HCN. Siderophore production in iron stress conditions provides rhizobia an added advantage, resulting in exclusion of pathogens due to iron starvation.