发生和未发生连作芦蒿枯萎病土壤的尖孢镰刀菌数量、产毒能力和致病力

尖孢镰刀菌引起的枯萎病是芦蒿连作障碍主要病害.对芦蒿不同种植年限发病和未发病土壤中尖孢镰刀菌数量、产毒素(镰刀菌酸)能力、致病力等开展研究.对采集的11份土壤样品分析结果显示:相较于不发病土壤,发病率≥40%的土壤尖孢镰刀菌数量显著增加;不发病和发病率≤25%的土壤尖孢镰刀菌数量没有显著差异(P>0.05).发病率为85%和40%的土壤尖孢镰刀菌高产毒素(镰刀菌酸产量>500μg L-1)的菌株比例显著(P<0.05)高于发病率为11%和不发病土壤.致病力测定显示发病率为85%和40%土壤中高致病力菌株比例显著(P<0.05)高于发病率为11%的土壤和不发病土壤,不致病菌株比例显著(P<0.05)低于发病率为11%的土壤和不发病土壤.土壤尖孢镰刀菌数量与连作年限没有显著相关性(P>0.05).发病土壤中,发病率、尖孢镰刀菌高产毒素菌株比例和高致病力菌株(病情指数>2)比例与连作年限显著(P<0.05)相关;未发病土壤中,尖孢镰刀菌高产毒素菌株比例和高致病力菌株比例与连作年限没有显著相关性(P>0.05).随着连作芦蒿发病率的增加,土壤中尖孢镰刀菌菌株数量、高产毒素菌株比例、致病菌株比例均显著增加(P<0.05),未发病连作土壤尖孢镰刀菌菌株数量、高产毒素菌株比例、致病菌株比例均没有显著变化.本研究结果可为连作芦蒿枯萎病发病机理的研究及其防治提供理论基础.     

八种镰刀菌毒素的电子捕获气相色谱分析

通过对各种反应条件的选择,建立了八种主要镰刀菌毒素同时检测的电子捕获气相色谱分析方法(GC-ECD法).在SE-54涂布的弹性石英毛细管交链柱上,八种镰刀菌毒素达到完全分离。T-2毒素,HT-2毒素、新茄病镰刀菌烯醇、二乙酰蘸草镰刀菌烯醇,玉米赤霉烯酮等的最低检测限为5×10~(-11)g,脱氧雪腐镰刀菌烯醇、雪腐镰刀菌烯醇、镰刀菌烯酮-X的最低检测限为3×10~(-12)g.     

脱氧雪腐镰刀菌烯醇、黄曲霉毒素B_1和玉米赤霉烯酮对秀丽隐杆线虫的联合毒性研究

脱氧雪腐镰刀菌烯醇(DON)、黄曲霉毒素B_1(AFB_1)和玉米赤霉烯酮(ZEN)是最为常见的粮食真菌毒素,易共存于谷物产品和动物饲料中,而目前对其联合毒性的研究较少,且研究结果并不完全一致。为探究DON、AFB_1和ZEN的联合毒性作用,本论文以秀丽隐杆线虫(C.elegans)为模型,分别评估了毒素混合物AFB_1+DON、AFB_1+ZEN、DON+ZEN和AFB_1+DON+ZEN对C.elegans的生长发育(体长)和生殖能力(产卵量)的毒性作用,并用Chou-Talalay模型来判定毒素混合物的相互作用类型。研究表明,AFB_1、DON和ZEN单独染毒C.elegans时,其毒作用强弱为AFB_1ZENDON。联合染毒时,AFB_1+DON对C.elegans产生协同作用,而DON+ZEN则产生拮抗作用;AFB_1+ZEN对体长(24 h)和产卵量的毒作用随着暴露浓度的增加,由弱拮抗变为协同作用,而在毒素暴露48 h后,对线虫的生长发育呈协同作用;AFB_1+DON+ZEN除在EC50-24 h和EC75-24 h时对体长产生明显的毒性增强作用外,其他普遍表现出拮抗作用。由此表明,DON、AFB_1和ZEN对C.elegans的联合毒性作用与剂量和时间相关。     

产类胡萝卜素菌株的筛选及其培养条件初探

从土壤、植物落花、细菌污染的组织培养二三角瓶里的培养基上及被细菌污染的LB琼脂平板上分离筛选出十几株产类胡萝卜素的菌株，其中从污染的LB琼脂平板上分离的一株菌落颜色为黄色的菌株PBH，分类鉴定为库特氏菌属(Kurthia sp.)．菌株在以葡萄糖为碳源，添加番茄汁(3．6mL／L)、芝麻油(1．6mL／L)、Na2CO3(6g/L)和磷酸缓冲盐的液体培养基中28℃振荡培养5d，每mL培养液细胞生物量湿重达到65．57mg，类胡萝卜素产量达到9．896μg，菌体每g湿重细胞胡萝卜素含量为150．9μg．图10参13     

重金属对禾谷镰刀菌的生长及毒素合成的影响

以禾谷镰刀菌为研究对象,研究了常见重金属(Cu、Pb、Zn、Cd)对禾谷镰刀菌菌株生长及其毒素合成的影响。研究结果表明,重金属对菌株的生长和其产毒能力均产生影响。Cu~(2+)和Cd~(2+)对菌株生长影响较大,随着浓度的增加对生长的抑制作用增强,当离子浓度分别为20 mg·L~(-1)和40 mg·L~(-1)时,能够完全抑制菌株生长。Zn~(2+)在0~160 mg·L~(-1)浓度范围内促进菌株生长,在10 mg·L~(-1)浓度下促进毒素合成。Pb~(2+)在察氏培养基中对菌株生长的影响没有明显规律可循,但是,随着Pb~(2+)浓度增加,抑制毒素合成作用增强。     

脱氧雪腐镰刀菌烯醇毒理作用研究进展

脱氧雪腐镰刀菌烯醇(DON)是人畜食品中检出率和超标率最高的一种真菌毒素。与核糖体60s亚基的肽基转移酶活性中心结合后会引发核糖体应激反应,诱导细胞凋亡等作用,对肠道、免疫系统造成破坏,并且具有遗传毒性。为今后的研究工作提供参考,笔者将从DON的毒理学机制和对机体的影响角度入手对近几年新的研究成果做综合论述。     

链格孢菌毒素细交链孢菌酮酸对蚕豆根尖细胞微核和有丝分裂的影响

利用蚕豆根尖为材料,研究了链格孢菌毒素细交链孢菌酮酸对细胞微核和有丝分裂的影响,以评价其细胞遗传毒性.结果表明,当浓度为12.5～50μg/mL时,细交链孢菌酮酸对蚕豆根尖细胞微核率和有丝分裂指数不产生显著影响,微核率低于3‰,有丝分裂指数超过9.5%.浓度升高到100～400μg/mL时,细交链孢菌酮酸使蚕豆根尖细胞微核率显著升高,有丝分裂指数显著下降,其中,400μg/mL时,微核率高达36.01‰,有丝分裂指数仅为1.90%.当细交链孢菌酮酸浓度为12.5～100μg/mL时,与对照相比,蚕豆根尖细胞多核仁率无显著差异,多核仁率低于3‰;浓度升高到200～400μg/mL时,细胞多核仁率显著升高,400μg/mL时,多核仁率高达26.10‰.     

不对称还原α-氨基苯乙酮菌种的筛选及转化条件优化

芳香基手性胺醇是许多手性药物合成的重要手性砌块,生物催化不对称还原前手性酮是合成该类醇的重要方法之一.以α-氨基苯乙酮盐酸盐为模型底物从土壤中筛选获得两株能分别高立体选择性催化底物产生R型、S型相应醇的菌株,对映体过量值(e.e.)分别为99%和77%,编号为1403和4802,鉴定菌株所属为镰刀菌属和地霉属.对两株菌培养时期和转化条件的研究表明镰刀菌1403最适生长时间为24 h,最优菌体浓度20 g/L,最优底物浓度5 g/L;地霉4802最适生长时间24 h,最优菌体浓度80 g/L,最优底物浓度3 g/L.底物特异性研究表明,菌株1403和4802均可转化α-氯代苯乙酮、α-溴代苯乙酮、α-羟基苯乙酮和苯乙酮为相应醇,且以α-羟基苯乙酮为底物时,其产物均为S型,e.e.值达99%.     

粮食与饲料中七种单端孢霉烯族真菌毒素的测定

本文提出了一种同时检测粮食和饲料中七种主要单端孢霉烯族真菌毒素的电子捕获气相色谱分析方法.粮食样品经醋酸乙酯、氯仿、乙醇提取后,减压浓缩.浓缩液用活性炭-中性氧化铝(2:1,W/W)柱净化,84％乙腈水溶液洗脱.洗脱液减压浓缩至于,残渣用N-七氟正丁酰咪唑衍生化.电子捕获气相色谱的分析结果表明,活性炭-中性氧化铝柱能有效地除去粮食中的干扰物,有很好的分离效果.T-2毒素、HT-2毒素、二乙酰藨草镰刀菌烯醇、新茄病镰刀菌烯醇、脱氧雪腐镰刀菌烯醇、雪腐镰刀菌烯醇、镰刀菌烯酮-X等七种毒素在小麦、大麦、玉米及大米中的回收率在浓度范围为50ng/g—500ng/g的范围内均为80％以上.     

粉葛根腐病的病原鉴定

近年来发现一种严重为害粉葛块根的土传病害。经显微镜直接检查、病原菌分离培养观察、致病性测定及田间病害调查，初步鉴定该病为粉葛镰刀菌根腐病，病原菌为腐皮镰刀菌(Fusarium solani，属半知菌亚门真菌)；该病的发生是由于腐皮镰刀菌与植物线虫复合感染所致。该报道在中国尚属首例。     

Clinical confirmation of trichothecene mycotoxicosis in patient urine

The investigations of four Cases involving mold-contaminated buildings and human reaction to exposure, documents tests of extracted urine containing trichothecene mycotoxins confirming exposure and the diagnosis of mycotoxicosis in humans. In each of four Cases, the urine demonstrated antibiotic activity, sulfuric acid charring, and protein release. Urine was extracted using ethyl acetate 40V/60V[EA]. Extracted mycotoxin spotted on (TLC) displayed color and a range of (rf) between 0.2-0.6 using various solvents. Extract was re-suspended using 50% ethanol V/V to inject mycotoxins into weanling female Sprague-Dawley rats. Degeneration and necrosis of the rat's tissue followed. Koch's Postulates conditions were fulfilled by isolation of the causative agent, the trichothecene mycotoxins and the reproduction of disease. Examination of human tissue within the urine extraction group confirms Koch's Postulates and comparative pathology confirms inhalation Mycotoxicosis, with severe necrosis of the central nervous system and severe scarring within the lungs. Extraction of mycotoxins from human patient urine is a very useful confirmatory test to demonstrate exposure and identify mycotoxicosis. Low concentrations (6%) of sodium hypochlorite were ineffective against the activity of trichothecene mycotoxin. The severity or stages of disease directly correlates the level of exposure or poisoning (Patent Pending).     

Fungal toxicity with special reference to mycotoxins

Mycotoxins are fungal secondary metabolites formed by consecutive series of enzyme-catalysed reactions from a few biochemically simple intermediates of primary metabolism. These mycotoxins can enter the human and animal food chain by direct or indirect contamination. Mycotoxins are equally harmful to animal and human beings. Realizing the importance of mycotoxins to the health of man and animals there have been concentrated efforts to develop highly sensitive analytical methods for detection and proper determination of mycotoxins in food, mixed feeds and feed ingredients, animal tissue, blood, urine and milk. Most of the mycotoxins are identified and most current research on it is concentrated on increasing sensitivity accuracy and reproducibility and above all to decrease the time of determination. A detailed review of mycotoxin and their detection is summarised in the paper.     

Die Bedeutung von Lebensmitteln tierischer Herkunft für die Mykotoxinaufnahme beim Menschen

Mycotoxins are a heterogeneous group of secondary fungal metabolites. Their formation in food and feedstuffs is influenced by many factors, including humidity, temperature, pH, oxygen concentration, type of substrate or presence of competitive microflora. The Food and Agriculture Organisation of the United Nations estimated in 1985 that approximately 25% of the world’s grain supply is contaminated with mycotoxins. There are more than 300 known mycotoxins; however, due to their occurrence and concentrations in food, and their toxic potential, only few of these are relevant with regard to consumer protection. These include the aflatoxins, ochratoxins and trichothecenes like deoxynivalenol, zearalenone and fumonisins. In Germany, only aflatoxins in food and feedstuffs are subjected to legal regulations. Beside the negative effects of mycotoxins on health and performance of farm animals, it is of importance to consider to what extent mycotoxins might be carried over into edible tissues like meat, milk and eggs when fed to farm animals. The present review shows that the carry over of mycotoxins into edible tissues is relatively low and is dependent on the specific mycotoxin and animal species. For example, due to the microbial decomposition of mycotoxins in the rumen, the carry over of mycotoxins into edible tissues of ruminants is considerably lower compared to monogastric species (pigs, chickens). Furthermore, after a short withdrawal period of 4 to 14 days combined with the feeding of non-contaminated diets, most of the mycotoxins, aside from ochratoxin A, are no longer detectable in any edible tissue. Ochratoxin A has a high binding affinity to specific blood proteins and is reabsorbed in the kidney; these mechanisms delay its elimination and increase the withdrawal period to at least 4 weeks. However, aside from the concentration of these substances in food, the daily intake of contaminated food is important for the risk analysis of mycotoxins to humans. In a recent study, it has been shown that more than 50% of the daily intake of ochratoxin A is derived from cereals and cereal products, and 30% from red wine, coffee and beer. Mear products from pigs and chickens only account for 4% of the total intake, while products from ruminants are considered to be negligible. These results indicate that mycotoxin-contaminated edible tissue, as compared to plant products, only plays a minor role with respect to consumer protection.     

Use of high-performance liquid chromatography to investigate the production of paralytic shellfish toxins by Protogonyaulax spp. in culture

Procedures have been developed for the extraction and high-performance liquid chromatography (HPLC) analysis of paralytic shellfish poisoning (PSP) toxins from Protogonyaulax spp. grown in batch culture. Using these procedures, the toxin content of two isolates of P. tamarensis (NEPCC 183 and 255) and one isolate of P. catenella (NEPCC 355) were examined. Total toxin and individual toxin concentrations were measured for each isolate during the exponential and stationary phases of growth in batch culture. The total toxicity of each isolate as measured by HPLC analysis was found to agree with toxicity as determined by the standard mouse bioassay. Two of the isolates (255 and 355) were found to be toxic and the third (183) was non-toxic. The toxic isolates (255 and 355) both showed higher average total PSP toxin content during the exponential phase (35 and 23 fmol toxin cell^-1, respectively) than during the stationary phase (21 and 8 fmol toxin cell^-1, respectively). These cultures differed dramatically in their toxin composition. P. tamarensis (255) contained a large proportion of the N(21) sulfo toxins (B₁, B₂, C₁, C₂) while P. catenella (355) contained primarily Gonyautoxins 1 through 4. The percent composition of individual toxins was found to be constant throughout the growth cycle for both toxic isolates, even though the total toxin concentration varied. Our results suggest that PSP toxin profiles might be useful as chemotaxonomic indicators.     

Plant diversity, CO2, and N influence inorganic and organic N leaching in grasslands

In nitrogen (N)-limited systems, the potential to sequester carbon depends on the balance between N inputs and losses as well as on how efficiently N is used, yet little is known about responses of these processes to changes in plant species richness, atmospheric CO2 concentration ([CO2]), and N deposition. We examined how plant species richness (1 or 16 species), elevated [CO2] (ambient or 560 ppm), and inorganic N addition (0 or 4 g x m(-2) x yr(-1)) affected ecosystem N losses, specifically leaching of dissolved inorganic N (DIN) and organic N (DON) in a grassland field experiment in Minnesota, USA. We observed greater DIN leaching below 60 cm soil depth in the monoculture plots (on average 1.8 and 3.1 g N x m(-2) x yr(-1) for ambient N and N-fertilized plots respectively) than in the 16-species plots (0.2 g N x m(-2) x yr(-1) for both ambient N and N-fertilized plots), particularly when inorganic N was added. Most likely, loss of complementary resource use and reduced biological N demand in the monoculture plots caused the increase in DIN leaching relative to the high-diversity plots. Elevated [CO2] reduced DIN concentrations under conditions when DIN concentrations were high (i.e., in N-fertilized and monoculture plots). Contrary to the results for DIN, DON leaching was greater in the 16-species plots than in the monoculture plots (on average 0.4 g N x m(-2) x yr(-1) in 16-species plots and 0.2 g N x m(-2) x yr(-1) in monoculture plots). In fact, DON dominated N leaching in the 16-species plots (64% of total N leaching as DON), suggesting that, even with high biological demand for N, substantial amounts of N can be lost as DON. We found no significant main effects of elevated [CO2] on DIN or DON leaching; however, elevated [CO2] reduced the positive effect of inorganic N addition on DON leaching, especially during the second year of observation. Our results suggest that plant species richness, elevated [CO2], and N deposition alter DIN loss primarily through changes in biological N demand. DON losses can be as large as DIN loss but are more sensitive to organic matter production and turnover.     

热带滨海区不同土地利用背景下土壤溶解性有机氮的组成与粒径分布特征

溶解态有机氮(DON)是土壤中活跃的氮库,其生态环境行为与它的化学组成和粒径分布密切相关。为评估热带滨海区不同土地利用方式对不同粒径中土壤溶解性有机氮组成特征的影响,从水稻田、橡胶园、菜园和果园采集土壤样品,通过一系列微滤和超滤(0.7,0.45,0.2,0.1μm,100,10,1 kDa)对土壤溶解性有机氮分级,并使用连续流动分析仪、三维荧光光谱和红外光谱研究了滤液中溶解态有机氮、无机氮的含量及荧光组分和有机官能团特征。结果表明,4种土地利用背景下土壤DON值的范围为5.25-10.88 mg·kg^-1,其大小顺序为水稻>菜园>果树>橡胶,且DON与溶解性总氮(DTN)的比值范围为26.08%-67.11%,其中橡胶土最高,水稻土最低;不同粒径下4种土地利用类型土壤DON主要集中在<100 kDa的粒径中,其值范围为4.85-9.48 mg·kg^-1,占全量的85.89%-92.41%。三维荧光光谱(3D-EEMs)及平行因子分析表明,4种土地利用背景下土壤DON含有两种类腐殖质组分及一种类蛋白质组分,且以类腐殖质组分为主,占比54.00%-77.67%;类蛋白组分对土地利用变化敏感,且随着粒径的减小,类蛋白组分占比增加,在<1 kDa组分中比例最高。红外光谱结果表明,4种土地利用背景下土壤DON主要在6个位置有相似的吸收峰,包含3410 cm^-1、1636 cm^-1、1402 cm^-1、1138-1035 cm^-1、673 cm^-1、602 cm^-1,不同土地利用背景下各吸收峰的透光度不同,强度最大的吸收来自游离的胺类N-H伸缩振动;水稻、菜园土壤DON芳香物质含量较高,结构较复杂。了解DON的组成与粒径分布对土地利用的响应,对进一步研究其生态环境行为具有重要意义。     

T-2毒素的毒性效应及致毒机制研究进展

T-2毒素是由多种镰刀菌产生的单端孢霉烯族化合物中毒性较强的一种毒素,广泛存在于谷物和谷类食品中.经食物连续摄入T-2毒素会对人类和动物的健康造成潜在危害.目前,T-2毒素对人和动物健康的影响受到了社会的广泛关注,已成为毒理学领域研究的热点课题之一.在综述了T-2毒素的生物转化、代谢途径及其毒理学效应与机制基础上,对现...     

Production of an antimicrobial cytochalasan by an endophytic Chaetomium globosum HYML55 from Hypericum mysorense and its RNA secondary structure analysis

The mutualistic benefits and disadvantages of non-clavicipitaceous endophytes for their hosts are not well studied and are postulated as being distinct from free-living or pathogenic counterparts. The defensive metabolites and antioxidants produced by endophytes are postulated to be the benefits offered for the hosts. Therefore, the antimicrobial and free-radical-scavenging activities of a foliar endophyte Chaetomium globosum HYML55 isolated from Hypericum mysorense of Western Ghats, India were studied. RNA secondary structure analysis refuted the unique biotope postulation because strain HYML55 was more similar to a coprophilous strain than to endophytic strains. Ethyl acetate extract of the fungus inhibited bacteria and fungi. The extract also quenched 98.65% 1,1-diphenyl- 2-picrylhydrazyl radicals with total antioxidant capacity of 13.62 mg ascorbic acid equivalent/g. A yellow amorphous antimicrobial compound purified from the extract was identified as chaetoglobosin F, which had a minimum inhibitory concentration of 7.8–15.6 μg/mL, except against Pseudomonas aeruginosa. This is the first report on the antimicrobial activity of chaetoglobosin F and other bioactivities of any endophytes from H. mysorense. The study emphasises the inter-habitat cycling of endophytes and possible benefits of C. globosum for the host. Chaetoglobosin F is an anti-inflammatory and anti-neoplastic compound, and C. globosum HYML55 could be exploited for the industrial production of this compound.