Production of Trichoderma strains with pesticide-polyresistance by mutagenesis and protoplast fusion

The sensitivity of two cold-tolerant Trichoderma strains belonging to the species T. harzianum and T.␣atroviride was determined to a series of pesticides widely used in agriculture. From the 16 pesticides tested, seven fungicides: copper sulfate, carbendazim, mancozeb, tebuconazole, imazalil, captan and thiram inhibited colony growth of the test strains significantly with minimal inhibitory concentrations of 300, 0.4, 50, 100, 100, 100 and 50 g/ml, respectively. Mutants resistant to carbendazim and tebuconazole were produced from both wild type strains by means of UV-mutagenesis. The cross-resistance capabilities and in␣vitro antagonistic properties of the mutants were determined. Carbendazim-resistant mutants showed total cross-resistance to benomyl and thiabendazole at a concentration of 20 g/ml. Intraspecific protoplast fusion was carried out between carbendazim- and tebuconazole-resistant mutants of both parental strains, and putative haploid recombinants with stable resistance to both pesticides were produced in the case of T.␣atroviride. These pesticide-polyresistant progenies are potential candidates for application in an integrated pest management system.This work was presented as an oral lecture in section ‘Agriculture, Soil, Forest Microbiology’ at the BioMicroWorld2005 conference.     

磁性纳米材料的生物医学应用进展

以四氧化三铁为代表的医用磁性纳米材料具有独特的磁学性能、表面易功能化、良好的生物学相容性等特点,在纳米医学相关领域展现出巨大的应用前景,特别是近年来它作为可介导外场的智能材料,在材料设计和生物医学应用方面均取得了突破性的进展.鉴于此,本文围绕磁性氧化铁纳米材料的生物医学应用,着重介绍近年来其在磁共振影像探针、磁热和磁力效应的生物医学应用、诊疗一体化以及纳米酶催化等领域的研究进展,并对磁性纳米材料在生物医学领域未来的发展方向进行了展望.     

基因工程FR-008/杀念菌素脱羧衍生物CS103的分离提取及毒性和抗白色念珠菌活性研究

本文在建立了基因工程FR-008/杀念菌素脱羧衍生物CS103分离提取工艺的基础上, 经进一步纯化, 获得一定供试样品。通过对比脱羧FR-008/杀念菌素CS103、FR-008/杀念菌素和两性霉素B三种化合物对人胚肾细胞毒性、对人血红细胞的溶血活性和对白色念珠菌的抗菌效果, 发现脱羧FR-008/杀念菌素CS103的毒性较FR-008/杀念菌素和两性霉素B大大降低, 且保持了较高的抗真菌(Candida albicans)活性。     

Antiviral effects of two Ganoderma lucidum triterpenoids against enterovirus 71 infection

Enterovirus 71 (EV71) is a major causative agent for hand, foot and mouth disease (HFMD), and fatal neurological and systemic complications in children. However, there is currently no clinical approved antiviral drug available for the prevention and treatment of the viral infection. Here, we evaluated the antiviral activities of two Ganoderma lucidum triterpenoids (GLTs), Lanosta-7,9(11),24-trien-3-one,15;26-dihydroxy (GLTA) and Ganoderic acid Y (GLTB), against EV71 infection. The results showed that the two natural compounds display significant anti-EV71 activities without cytotoxicity in human rhabdomyosarcoma (RD) cells as evaluated by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) cell proliferation assay. The mechanisms by which the two compounds affect EV71 infection were further elucidated by three action modes using Ribavirin, a common antiviral drug, as a positive control. The results suggested that GLTA and GLTB prevent EV71 infection through interacting with the viral particle to block the adsorption of virus to the cells. In addition, the interactions between EV71 virion and the compounds were predicated by computer molecular docking, which illustrated that GLTA and GLTB may bind to the viral capsid protein at a hydrophobic pocket (F site), and thus may block uncoating of EV71. Moreover, we demonstrated that GLTA and GLTB significantly inhibit the replication of the viral RNA (vRNA) of EV71 replication through blocking EV71 uncoating. Thus, GLTA and GLTB may represent two potential therapeutic agents to control and treat EV71 infection.     

The potency of lemon (Citrus limon L.) essential oil to control some fungal diseases of grapevine wood

This study aimed to evaluate the in vitro antifungal activity (AA) of the essential oil (EO) of lemon (Citrus limon L.) against three pathogenic fungi attacking grapevine wood. The composition of the EO was also studied. Ten volatile components were identified by gas chromatography–mass spectrometry. The results showed that the EO consists of volatile components where monoterpene hydrocarbons are the most abundant ones. Four major components were identified, which represent 99.9% of the total EO (limonene, neral, ß-pinene, and ?-terpinene). The AA of the EO was evaluated against three pathogenic fungi attacking grapevine wood (Eutypa sp., Botryosphaeria dothidea, and Fomitiporia mediterranea). The results showed that the EO exerts AA against all tested fungi and significantly inhibits their growth. Eutypa sp. is the most sensitive fungus. These results show, for the first time, a new use for the EO of lemon (C. limon L.) to control fungal diseases of grapevine wood.     

Investigating the bioactivity of cells and cell-free extracts of Streptomyces griseus towards Fusarium oxysporum f. sp. cubense race 4

The antifungal activity of viable cells of Streptomyces griseus (St 4) and its cell-free extracts were investigated against the pathogenic Fusarium oxysporum f. sp. cubense race 4 (FOC race 4), causal agent of wilt disease in bananas. Results from in vitro and soil assays showed cells and cell-free extracts of S. griseus were able to inhibit FOC race 4 with varying degree of success. Antifungal activity was attributed to chitinase and β-1,3-glucanase, detected in both cells and cell-free extracts, which caused lysis of fungal cell wall and inhibited sporulation. Interestingly, β-1,3-glucanase and chitinase activities were significantly higher in cell-free extracts compared to cells, with 8.30 and 5.43 against 7.96 and 4.95 U mL⁻¹, respectively. Application to soil however, showed inoculation using S. griseus cells were more effective in suppressing growth of FOC race 4 than crude extracts, with 6 log₁₀ CFU of FOC race 4 g⁻¹ soil enumerated compared to 7 log₁₀ CFU of FOC race 4 g⁻¹ soil after 20 days. To summarize, this study has shown that cell-free extracts of S. griseus have antifungal properties but may not be suitable for soil application in its current form (liquid suspension). Further investigations on bioformulation may address this limitation.     

Anticandidial and anticryptococcal activity of <Emphasis Type="Italic">Euphorbia fusiformis</Emphasis>, a rare medicinal plant

The present work demonstrates the screening of extracts of the rare medicinal herb Euphorbia fusiformis for antifungal activity. The main aim was to investigate its antifungal properties against Candida albicans and Cryptococcus neoformans, the causative agents of human candidiasis and cryptococcosis, respectively. Aqueous and organic solvent extracts from the leaves and rootstock of the plant were tested against the fungi by the well-in-agar method. Almost all the organic solvent extracts exhibited an inhibitory effect against C. albicans and to some extent on C. neoformans, except for the aqueous extracts, which had no effect. The combined formulations of the extracts also had better activity against C. albicans than C. neoformans. This study thus concludes by demonstrating the antifungal properties of E. fusiformis and also the potential research in identifying the active principles, which may have future therapeutic value.     

260例浅部真菌病及致病菌种分析

目的了解新疆乌鲁木齐市浅部真菌病发病情况及其病原菌的菌种分布。方法对2004年2月至2010年08月在新疆医科大学第一附属医院皮肤科门诊就诊,临床拟诊为头癣、甲癣、体股癣、手足癣的1 308例患者取病发、皮屑、甲屑行真菌镜检、培养。培养阳性标本在形态学鉴定的基础上行核糖体DNA(rDNA)ITS区序列测定确切鉴定菌种。使用SPSS 17.0统计软件对于结果进行统计分析。结果真菌镜检培养均为阳性患者260例,头癣培养阳性率最高,占33.1%,其次为体股癣占28.8%、甲癣占21.5%、手足癣占16.5%;菌种鉴定:红色毛癣菌为最常见的致病菌(28.5%),其次为须癣毛癣菌(22.3%)、犬小孢子菌(18.8%)、断发毛癣菌(10.0%)。犬小孢子菌为头癣最主要致病菌(16.2%)。统计学分析显示:不同性别、族别间体股癣、手足癣发病率有统计学意义(P<0.05),均好发于男性,汉族发病率高于维吾尔族。不同年龄头癣、甲癣、体股癣、手足癣发病率差异也均有统计学意义(P<0.05),甲癣、体股癣、手足癣好发于中青年,头癣好发于儿童。结论乌鲁木齐市浅部真菌病主要致病真菌是红色毛癣菌。头癣是本地区最主要的儿童浅部真菌病,主要病原菌为犬小孢子菌。     

循环水养殖系统中凡纳滨对虾肠道微生物对三种复合益生菌制剂的响应

【目的】在循环养殖系统中应用不同的复合益生菌制剂,探讨凡纳滨对虾肠道菌群结构特征及免疫水平发生的变化。【方法】30 d养殖周期结束后,通过平板计数法分析肠道细菌总数和弧菌总数;基于高通量测序技术分析肠道样品V3+V4区菌群特征;采用实时荧光定量PCR技术分析免疫相关因子TLR1和Dorsal基因表达水平,阐述益生菌制剂应用的意义。【结果】益生菌制剂的应用降低了凡纳滨对虾肠道中细菌总数,抑制弧菌的生长,间接预防疾病的发生。不同益生菌制剂从不同程度上优化了凡纳滨对虾肠道菌群结构,提高高质量序列和有效OTU数量,Chao1、Simpson、Shannon指数显示了丰富度和多样性变化,复合益生菌制剂3效果较好。同时,菌群结构得到优化,其中益生菌制剂1组对拟杆菌门含量百分比产生显著影响。Toll受体TLR1和Toll通路中的Dorsal基因m RNA表达受到益生菌制剂的影响,1、3组促进了TLR 1表达,1、2、3组都促进了Dorsal基因表达。【结论】在循环水养殖系统中添加益生菌制剂可优化凡纳滨对虾肠道菌群特征,提高免疫水平,为病害防控和健康养殖提供理论依据。     

Pyrrolomycins as potential anti-staphylococcal biofilms agents

With the goal of discovering new anti-infective agents active against microbial biofilms, this investigation focused on some natural pyrrolomycins, a family of halogenated pyrrole antibiotics. In this study the anti-staphylococcal biofilm activity of pyrrolomycins C, D, F1, F2a, F2b, F3 and of the synthesized related compounds I, II, III were investigated. The susceptibility of six staphylococcal biofilms was determined by methyltiazotetrazolium staining. Most of the compounds were active at concentrations of 1.5 μg ml^?1 with significant inhibition percentages. A few of the compounds were active at the lowest screening concentration of 0.045 μg ml^?1. The population log reduction of activity against the two best biofilm forming Staphylococcus aureus strains as determined by viable plate counts is also reported. In order to adequately assess the utility of these compounds, their toxicity against human cells was evaluated. It is concluded that pyrrolomycins and synthetic derivatives are promising compounds for developing novel effective chemical countermeasures against staphylococcal biofilms.