Antifungal activity of chitosan against Fusarium oxysporum f. sp. cubense

The in vitro antifungal activity of chitosan against Fusarium oxysporum f. sp. cubense Race 4 (FocR4) the causal agent of banana wilt was investigated. Chitosan at all concentrations tested reduced the hyphal growth of FocR4 on potato dextrose agar media and recording maximum inhibition of 76.36% at 8 mg/mL. The inhibitory effect was found to increase as chitosan concentration increases. The 50% effective concentration value was estimated by probit analysis, and it was 1.4 mg/mL. Chitosan was more effective in potato dextrose broth where it completely inhibited the mycelial growth of FocR4 at all concentrations tested. Chitosan inhibited the sporulation of FocR4 by a maximum of 96.53% at 8 mg/mL chitosan, and 100% inhibition for spore germination was recorded at all concentrations tested. Chitosan at concentrations of more than 1.6 mg/mL was also found to induce morphological changes in FocR4 characterized by agglomeration of hyphae, abnormal shapes, vesicles, or empty cells devoid of cytoplasm in the mycelia. © 2010 Wiley Periodicals, Inc. J Appl Polym Sci, 2011     

Control of Panama Disease of Banana by Rotating and Intercropping with Chinese Chive (Allium Tuberosum Rottler): Role of Plant Volatiles

Intercropping and rotating banana (Musa spp.) with Chinese chive (Allium tuberosum Rottler) has been used as an effective method to control Panama disease (Fusarium wilt) of banana in South China. However, the underlying mechanism is unknown. In this study, we used aqueous leachates and volatiles from Chinese chive to evaluate their antimicrobial activity on Fusarium oxysporum f. sp. cubense race 4 (FOC), the causal agent of Panama disease in banana, and identified the antifungal compounds. Both leaf and root leachates of Chinese chive displayed strong inhibition against FOC, but the concentrated leachates showed lower inhibition than the original leachates. In a sealed system volatiles emitted from the leaves and roots of Chinese chive inhibited mycelial growth of FOC. Volatile compounds emitted from the intact growing roots mimicking natural environment inhibited spore germination of FOC. We identified five volatiles including 2-methyl-2-pentenal and four organosulfur compounds (dimethyl trisulfide, dimethyl disulfide, dipropyl disulfide, and dipropyl trisulfide) from the leaves and roots of Chinese chive. All these compounds exhibited inhibitory effects on FOC, but 2-methyl-2-pentenal and dimethyl trisulfide showed stronger inhibition than the other three compounds. 2-Methyl-2-pentenal at 50–100 μl/l completely inhibited the mycelial growth of FOC. Our results demonstrate that antifungal volatiles released from Chinese chive help control Panama disease in banana. We conclude that intercropping and rotating banana with Chinese chive can control Panama disease and increase cropland biodiversity.     

八宝景天叶乙醇提取物的抑真菌活性

[方法]采用菌丝生长速率法研究了八宝景天叶乙醇提取物对9种植物病原真菌的抑制作用,同时采用菌丝生长速率和孢子萌发2种方法测定其对玉米弯孢病菌的抑菌活性。[结果]在质量浓度为200 g/L时,对9种植物病原真菌均有一定的抑制作用,且抑制率均达75%以上,其中对玉米弯孢病菌的抑制作用最强,抑制率为82.27%,对其孢子萌发的抑菌活性高于对其菌丝生长的抑菌活性,EC50值分别为74.18、66.53 g/L。[结论]八宝景天叶有着良好的抑菌效果,具有很好的开发价值。     

嘧菌酯对几种植物病原真菌的室内活性测定

试验结果表明,嘧菌酯对供试8种植物病原真菌具有很高的活性,既可抑制病菌菌丝生长又能抑制孢子萌发。水稻纹枯病菌的菌丝对嘧菌酯最敏感,EC50为0．0185mg／L;嘧菌酯对辣椒根腐病菌孢子萌发的抑制作用最明显,EC50达到0．0104mg／L。     

Isolation and Heterologous Expression of a Polygalacturonase Produced by Fusarium oxysporum f. sp. cubense Race 1 and 4

Fusarium wilt (Panama disease) caused by Fusarium oxysporum f. sp. cubense (FOC) represents a significant threat to banana (Musa spp.) production. Musa AAB is susceptible to Race 1 (FOC1) and Race 4 (FOC4), while Cavendish Musa AAA is found to be resistant to FOC1 but still susceptible to Race 4. A polygalacturonase (PGC3) was purified from the supernatant of Fusarium oxysporum f. sp. cubense race 4 (FOC4), which is the pathogen of Fusarium wilt. PGC3 had an apparent molecular weight of 45 kDa according to SDS-PAGE. The enzyme hydrolyzed polygalacturonic acid in an exo-manner, as demonstrated by analysis of degradation products. The K_m and V_max values of PGC3 from FOC4 were determined to be 0.70 mg·mL⁻¹ and 101.01 Units·mg·protein⁻¹·min⁻¹, respectively. Two pgc3 genes encoding PGC3 from FOC4 and FOC1, both genes of 1368 bp in length encode 456 amino-acid residues with a predicted signal peptide sequence of 21 amino acids. There are 16 nucleotide sites difference between FOC4-pgc3 and FOC1-pgc3, only leading to four amino acid residues difference. In order to obtain adequate amounts of protein required for functional studies, two genes were cloned into the expression vector pPICZaA and then expressed in Pichia pastoris strains of SMD1168. The recombinant PGC3, r-FOC1-PGC3 and r-FOC4-PGC3, were expressed and purified as active proteins. The optimal PGC3 activity was observed at 50 °C and pH 4.5. Both recombinant PGC3 retained >40% activity at pH 3–7 and >50% activity in 10–50 °C. Both recombinant PGC3 proteins could induce a response but with different levels of tissue maceration and necrosis in banana plants. In sum, our results indicate that PGC3 is an exo-PG and can be produced with full function in P. pastoris.     

放线菌发酵提取物对百合炭疽病的抗菌活性筛选

从云南不同生态环境中采集的土壤样品巾分离到334株放线菌,以百合炭疽病菌作为指示菌,通过菌株发酵提取物对孢子萌发抑制、菌丝生长抑制和温室盆栽防治效果的测定,初步筛选出对炭疽病菌分生孢子萌发有较强抑制能力的18个菌株,对炭疽病菌菌丝生长有较强抑制能力的24个菌株,离体叶片防治效果60％以上的14个菌株,其中YIM31927、YIM31047等10株发酵提取物对百合炭疽病菌孢子萌发、菌丝生长和离体叶片防治试验均表现出了一致活性,筛选到一批具有开发应用潜能的放线菌资源。     

Inhibition of Citrinin Production in Penicillium citrinum Cultures by Neem [Azadirachta indica A. Juss (Meliaceae)]

The efficacy of different concentrations of aqueous neem leaf extract (3.12 to 50 mg/mL) on growth and citrinin production in three isolates of Penicillium citrinum was investigated under laboratory conditions. Mycotoxin production by the isolates was suppressed, depending on the concentration of the plant extract added to culture media at the time of spore inoculation. Citrinin production in fungal mycelia grown for 21 days in culture media containing 3.12 mg/mL of the aqueous extract of neem leaf was inhibited by approximately 80% in three isolates of P. citrinum. High-performance liquid chromatography was performed to confirm the spectrophotometric results. Vegetative growth was assessed, but neem extract failed to inhibit it. Neem leaf extract showed inhibition of toxin production without retardation in fungal mycelia growth.     

The Effects of Nanomaterials as Endocrine Disruptors

In recent years, nanoparticles have been increasingly used in several industrial, consumer and medical applications because of their unique physico-chemical properties. However, in vitro and in vivo studies have demonstrated that these properties are also closely associated with detrimental health effects. There is a serious lack of information on the potential nanoparticle hazard to human health, particularly on their possible toxic effects on the endocrine system. This topic is of primary importance since the disruption of endocrine functions is associated with severe adverse effects on human health. Consequently, in order to gather information on the hazardous effects of nanoparticles on endocrine organs, we reviewed the data available in the literature regarding the endocrine effects of in vitro and in vivo exposure to different types of nanoparticles. Our aim was to understand the potential endocrine disrupting risks posed by nanoparticles, to assess their underlying mechanisms of action and identify areas in which further investigation is needed in order to obtain a deeper understanding of the role of nanoparticles as endocrine disruptors. Current data support the notion that different types of nanoparticles are capable of altering the normal and physiological activity of the endocrine system. However, a critical evaluation of these findings suggests the need to interpret these results with caution since information on potential endocrine interactions and the toxicity of nanoparticles is quite limited.     

New Antifungal Pyranoisoflavone from Ficus tikoua Bur

Considering the undesirable attributes of synthetic fungicides and the availability of Ficus species in China, the stem of Ficus tikoua Bur. was investigated. One new antifungal pyranoisoflavone, 5,3',4'-trihydroxy-2″,2″-dimethylpyrano (5″,6″:7,8) isoflavone (1), together with two known isoflavones, wighteone (2) and lupiwighteone (3) (with previously reported antifungal activities), were isolated from ethyl acetate extract by bioassay-guided fractionation. Their structures were determined by spectroscopic analysis, such as NMR ((1)H-(1)H COSY, HMQC, HMBC and NOESY), IR, UV and HRMS, as well as ESI-MS(n) analyses. The antifungal activities of 1-3 against Phytophthora infestans were evaluated by direct spore germination assay, and the IC(50) values were 262.442, 198.153 and 90.365 μg·mL(-1), respectively.     

Acute exposure to silica nanoparticles enhances mortality and increases lung permeability in a mouse model of Pseudomonas aeruginosa pneumonia

Background

The lung epithelium constitutes the first barrier against invading pathogens and also a major surface potentially exposed to nanoparticles. In order to ensure and preserve lung epithelial barrier function, the alveolar compartment possesses local defence mechanisms that are able to control bacterial infection. For instance, alveolar macrophages are professional phagocytic cells that engulf bacteria and environmental contaminants (including nanoparticles) and secrete pro-inflammatory cytokines to effectively eliminate the invading bacteria/contaminants. The consequences of nanoparticle exposure in the context of lung infection have not been studied in detail. Previous reports have shown that sequential lung exposure to nanoparticles and bacteria may impair bacterial clearance resulting in increased lung bacterial loads, associated with a reduction in the phagocytic capacity of alveolar macrophages.

Results

Here we have studied the consequences of SiO₂ nanoparticle exposure on Pseudomonas aeruginosa clearance, Pseudomonas aeruginosa-induced inflammation and lung injury in a mouse model of acute pneumonia. We observed that pre-exposure to SiO₂ nanoparticles increased mice susceptibility to lethal pneumonia but did not modify lung clearance of a bioluminescent Pseudomonas aeruginosa strain. Furthermore, internalisation of SiO₂ nanoparticles by primary alveolar macrophages did not reduce the capacity of the cells to clear Pseudomonas aeruginosa. In our murine model, SiO₂ nanoparticle pre-exposure preferentially enhanced Pseudomonas aeruginosa-induced lung permeability (the latter assessed by the measurement of alveolar albumin and IgM concentrations) rather than contributing to Pseudomonas aeruginosa-induced lung inflammation (as measured by leukocyte recruitment and cytokine concentration in the alveolar compartment).

Conclusions

We show that pre-exposure to SiO₂ nanoparticles increases mice susceptibility to lethal pneumonia but independently of macrophage phagocytic function. The deleterious effects of SiO₂ nanoparticle exposure during Pseudomonas aeruginosa-induced pneumonia are related to alterations of the alveolar-capillary barrier rather than to modulation of the inflammatory responses.

Electronic supplementary material

The online version of this article (doi:10.1186/s12989-014-0078-9) contains supplementary material, which is available to authorized users.     

Nanoparticle Encapsidation of Flock House Virus by Auto Assembly of Tobacco Mosaic Virus Coat Protein

Tobacco Mosaic virus (TMV) coat protein is well known for its ability to self-assemble into supramolecular nanoparticles, either as protein discs or as rods originating from the ~300 bp genomic RNA origin-of-assembly (OA). We have utilized TMV self-assembly characteristics to create a novel Flock House virus (FHV) RNA nanoparticle. FHV encodes a viral polymerase supporting autonomous replication of the FHV genome, which makes it an attractive candidate for viral transgene expression studies and targeted RNA delivery into host cells. However, FHV viral genome size is strictly limited by native FHV capsid. To determine if this packaging restriction could be eliminated, FHV was adapted to express enhanced green fluorescent protein (GFP), to allow for monitoring of functional FHV RNA activity. Then TMV OA was introduced in six 3'' insertion sites, with only site one supporting functional FHV GFP expression. To create nanoparticles, FHV GFP-OA modified genomic RNA was mixed in vitro with TMV coat protein and monitored for encapsidation by agarose electrophoresis and electron microscopy. The production of TMV-like rod shaped nanoparticles indicated that modified FHV RNA can be encapsidated by purified TMV coat protein by self-assembly. This is the first demonstration of replication-independent packaging of the FHV genome by protein self-assembly.     

Antifungal activity of denture soft lining material modified by silver nanoparticles-a pilot study

Soft liner materials in oral cavity environments are easily colonized both by fungi and dental plaque. These factors are the cause of mucosal infections. The microorganism that most frequently colonizes soft liner materials is Candida albicans. Colonization occurs on the surface of materials and within materials. A solution to this problem might involve modification of soft liner materials with silver nanoparticles (AgNPs). In this article, we present results showing the antifungal efficacy of silicone soft lining materials modified with AgNPs. The modification process was conducted by dissolving both material components (base and catalyst) in a colloidal solution of AgNPs and evaporating the solvent. Composites with various AgNP concentrations (10, 20, 40, 80, 120 and 200 ppm) were examined. The in vitro antifungal efficacy (AFE) of composite samples was 16.3% to 52.5%.     

Antioxidant Potential,Lipid Peroxidation Inhibition and Antimicrobial Activities of Satureja montana L. subsp. kitaibelii Extracts

The antioxidant activity of different Satureja montana L. subsp. kitaibelii extracts was tested by measuring their ability to scavenge reactive hydroxyl radical during the Fenton reaction, using ESR spectroscopy. Also, the influence of these extracts on lipid peroxyl radicals obtained during lipid peroxidation of: (I) sunflower oil (37 ºC, 3h) induced by 4,4′-azobis(4-cyanovaleric acid) (ACVA) and (II) liposomes induced by 2,2′-azobis(2- amidino-propane)dihydrochloride (AAPH) was studied. n-Butanol extract had the best antioxidant activity (100% at 0.5 mg/mL in Fenton reaction system; 89.21% at 5 mg/mL in system I; 83.38% at 5 mg/mL in system II). The antioxidant activities of the extracts significantly correlated with total phenolic content. The antimicrobial activity of Satureja montana L. subsp. kitaibelii extracts was investigated. Petroleum ether, chloroform and ethyl acetate extracts expressed a wide range of inhibiting activity against both grampositive and gram-negative bacteria.     

Antifungal Activity of Bacillus velezensis CE 100 against Anthracnose Disease (Colletotrichum gloeosporioides) and Growth Promotion of Walnut (Juglans regia L.) Trees

Walnut anthracnose caused by Colletotrichum gloeosporioides is a deleterious disease that severely affects the production of walnut (Juglans regia L.). The aim of this study was to assess the antifungal and growth promotion activities of Bacillus velezensis CE 100 as an alternative to chemical use in walnut production. The crude enzyme from B. velezensis CE 100 exhibited chitinase, protease, and β-l,3-glucanase activity and degraded the cell wall of C. gloeosporioides, causing the inhibition of spore germination and mycelial growth by 99.3% and 33.6% at 100 µL/mL, respectively. The field application of B. velezensis CE 100 culture broth resulted in a 1.3-fold and 6.9-fold decrease in anthracnose disease severity compared to the conventional and control groups, respectively. Moreover, B. velezensis CE 100 produced indole-3-acetic acid (up to 1.4 µg/mL) and exhibited the potential for ammonium production and phosphate solubilization to enhance the availability of essential nutrients. Thus, field inoculation of B. velezensis CE 100 improved walnut root development, increased nutrient uptake, enhanced chlorophyll content, and consequently improved total biomass by 1.5-fold and 2.0-fold compared to the conventional and control groups, respectively. These results demonstrate that B. velezensis CE 100 is an effective biocontrol agent against anthracnose disease and a potential plant growth-promoting bacteria in walnut tree production.     

植物源农药的筛选及补骨脂抑菌活性

以水稻稻瘟病菌、辣椒疫霉病菌、黄瓜炭疽病菌为靶标菌，通过孢子萌发试验对12种植物提取物的抑菌活性进行粗筛。结果表明，植物粗提物在100mg／L质量浓度下，补骨脂提取物对水稻稻瘟病菌和辣椒疫霉病菌孢子萌发的抑制率均为100％，虎杖和黄柏提取物对水稻稻瘟病菌孢子萌发的抑制率均为100％。然后，采用菌丝生长速率法将补骨脂提取物对11种植物病原菌的抑菌活性进行了测定，结果表明，补骨脂粗提物对苹果腐烂等11种植物病原菌均具有抑制作用，其中，对苹果腐烂病菌、黄瓜炭疽病菌、小麦赤霉病菌、水稻稻瘟病菌的EC50分别为17．669、73．136、77．294、32．141mg／L。     

Antifungal Activity of Essential Oil and Extracts of <Emphasis Type="Italic">Piper chaba</Emphasis> Hunter Against Phytopathogenic Fungi

The efficacy of the essential oil and various leaf extracts of Piper chaba Hunter was evaluated for controlling the growth of some important phytopathogenic fungi. The hydrodistilled essential oil was analysed by GC–MS. Fifty-three compounds representing 95.1% of the total oil were identified, of which α-humulene (16.4%), caryophyllene oxide (12.2%), viridiflorol (8.1%), globulol (7.4%), β-selinene (7.1%), spathulenol (6.2%), (E)-nerolidol (5.1%), linalool (4.5%) and 3-pentanol (3.5%) were the major compounds. The oil (1,000 ppm) and organic extracts (1,500 ppm) of P. chaba Hunter displayed a great potential of antifungal activity as a growth inhibition against the tested phytopathogenic fungi, Fusarium oxysporum KACC 41083, Phytophthora capsici KACC 40157, Colletotrichum capsici KACC 410978, Fusarium solani KACC 41092 and Rhizoctonia solani KACC 40111 in the range of 55.1–70.3% and minimum inhibitory concentration ranging from 125 to 500 μg/ml. The oil had also a strong detrimental effect on spore germination of all the tested fungi along with concentration as well as time-dependent kinetic inhibition of P. capsici. The results obtained from this study may contribute to the development of new antifungal agents to protect crops from fungal diseases.     

An in situ study on the coalescence of monolayer-protected Au-Ag nanoparticle deposits upon heating

The structural evolution of thiolate-protected nanoparticles of gold, silver, and their alloys with various Au/Ag ratios (3:1, 1:1, and 1:3) upon heating was investigated by means of in situ synchrotron radiation X-ray diffraction. The relationships between the coalescence and composition of nanoparticles, as well as the surfactant reactions, were clarified. Experimental results show that there existed a critical temperature ranging from 120°C to 164°C, above which the tiny broad X-ray diffraction peaks became sharp and strong due to particle coalescence. The coalescence temperatures for alloy nanoparticle deposits were clearly lower than those for pure metals, which can be ascribed to the rivalry between the thermodynamic effect due to alloying and the interactions between surface-assembled layers and the surface atoms of the nanoparticles. The strong affinity of thiolates to Ag and thus complex interactions give rise to a greater energy barrier for the coalescence of nanoparticles into the bulk and subsequent high coalescence temperature. The influences of particle coalescence on the optical and electrical properties of the nanoparticle deposits were also explored.     

Synthesis and antifungal activity of reduced graphene oxide nanosheets

Reduced graphene oxide (rGO) nanosheets were produced using a modified Hummers method. Antifungal activity of rGO nanosheets was tested against three fungi i.e., Aspergillus niger (A. niger), Aspergillus oryzae (A. oryzae) and Fusarium oxysporum (F. oxysporum). The rGO inhibits the mycelial growth of the fungi and it is believed that this is due to its sharp edges. The half maximal inhibitory concentration (IC₅₀), a measure of the effectiveness of the rGO in inhibiting the fungi, was investigated. IC₅₀ values of the rGO against F. oxysporum, A. niger, and A. oryzae are 50, 100, and 100 μg ml^?1, respectively.     

Synthesis of silver nanoparticles using reducing agents obtained from natural sources (Rumex hymenosepalus extracts)

We have synthesized silver nanoparticles from silver nitrate solutions using extracts of Rumex hymenosepalus, a plant widely found in a large region in North America, as reducing agent. This plant is known to be rich in antioxidant molecules which we use as reducing agents. Silver nanoparticles grow in a single-step method, at room temperature, and with no addition of external energy. The nanoparticles have been characterized by ultraviolet-visible spectroscopy and transmission electron microscopy, as a function of the ratio of silver ions to reducing agent molecules. The nanoparticle diameters are in the range of 2 to 40 nm. High-resolution transmission electron microscopy and fast Fourier transform analysis show that two kinds of crystal structures are obtained: face-centered cubic and hexagonal.     

Biogenic synthesis and characterization of gold nanoparticles by Escherichia coli K12 and its heterogeneous catalysis in degradation of 4-nitrophenol

Room-temperature extracellular biosynthesis of gold nanoparticles (Au NPs) was achieved using Escherichia coli K12 cells without the addition of growth media, pH adjustments or inclusion of electron donors/stabilizing agents. The resulting nanoparticles were analysed by ultraviolet–visible (UV–vis) spectrophotometry, atomic force microscopy, transmission electron microscopy and X-ray diffraction. Highly dispersed gold nanoplates were achieved in the order of around 50 nm. Further, the underlying mechanism was found to be controlled by certain extracellular membrane-bound proteins, which was confirmed by Fourier transformation-infrared spectroscopy and sodium dodecyl sulfate polyacrylamide gel electrophoresis. We observed that certain membrane-bound peptides are responsible for reduction and subsequent stabilization of Au NPs (confirmed by zeta potential analysis). Upon de-activation of these proteins, no nanoparticle formation was observed. Also, we prepared a novel biocatalyst with Au NPs attached to the membrane-bound fraction of E. coli K12 cells serving as an efficient heterogeneous catalyst in complete reduction of 4-nitrophenol in the presence of NaBH₄ which was studied with UV–vis spectroscopy. This is the first report on bacterial membrane-Au NP nanobiocomposite serving as an efficient heterogeneous catalyst in complete reduction of nitroaromatic pollutant in water.