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1.
紫花针茅作为青藏高原高寒草原群落中主要优势禾草之一,关于该植物所感染禾草内生真菌的形态和分类研究尚未见报道.本研究采用分离培养获取不同样点的紫花针茅内生真菌菌落,利用特异性引物克隆紫花针茅内生真菌序列,并与Genbank中下载的序列共同构建系统进化树.结果表明,青海省紫花针茅样品带菌率高达100%,而其他区域样品均不带菌.分离的内生真菌菌落从菌落形态,生长速度及分生孢子形态等特征均与Epichloё属内生真菌的形态特征相似.系统进化关系表明,它们分别与北美洲竖针茅体内无性态内生真菌Epichloё chisosa,醉马草内生真菌Epichloё inebrians及甘肃内生真菌Epichloё gansuensis具有较近的亲缘关系,进一步说明紫花针茅所感染内生真菌与宿主间未体现出严格的宿主特异性.  相似文献   

2.
异针茅作为青藏高原地区重要的乡土草种之一,是高寒草甸草原主要的建群禾草种类。迄今为止,有关异针茅所感染内生真菌的种类和分类地位的研究尚未见报道。基于此,以青海高原地区两个地理种群的异针茅为研究对象,采用常规分离培养形态观测和管家基因扩增目的片段基因并与已公布的内生真菌序列构建系统发育树的方法,确定异针茅感染内生真菌的种类和分类地位。结果表明:异针茅分离出的内生真菌菌落形态、生长速度和分生孢子形态等均与Epichloё属内生真菌的形态特征相似。编码蛋白tub和肌动蛋白actin序列构建的系统发育树表明,异针茅感染的内生真菌种类分别为Epichloёgansuensis和Epichloёsylvatica,而转录延长因子tef序列构建的系统发育树发现异针茅感染的内生真菌种类分别为Epichloёinebrians和E.sylvatica。由此可见,异针茅感染的内生真菌分别为甘肃内生真菌E.gansuensis、醉马草内生真菌E.inebrians和欧洲短柄草内生真菌E.sylvatica;进一步揭示异针茅感染的内生真菌表现出多样性,异针茅与其感染的内生真菌间未形成严格的宿主特异性。  相似文献   

3.
捕食线虫性真菌的分离培养技术与形态学   总被引:19,自引:3,他引:16  
分离培养捕食线虫性真菌的最佳培养基为 0 .4g/ L 玉米粉琼脂培养基 (CMA) ;0 .4g/ L 玉米粉液体培养基可作为保存菌种培养基。在分离培养过程中 ,必须加入活的线虫第 3期幼虫 ,以诱导菌株产生捕食性器官——菌环、菌网 ,并与其他真菌鉴别。捕食线虫性真菌有菌丝和孢子 2种形态 ,其发育时期不同。本试验分离的少孢节丛孢菌(Arthrobotrys oligospora) ,菌丝发达 ,无色有隔 ,隔距不等 ;分生孢子梗直立不分支 ,分生孢子围绕着分生孢子梗呈轮状排列 ,似梅花瓣状 ,2层以上 ;孢子呈倒卵形 ,丰满无色 ,有横隔 ,游离端大而钝圆 ,梗端小而稍尖。  相似文献   

4.
赵清梅  余永涛  贝盏临 《草业科学》2012,29(12):1821-1826
为研究苦马豆(Swainsona salsula)中内生真菌的种属分类,对采自宁夏的苦马豆内生真菌进行分离培养和形态观察,对分离菌株的ITS序列进行扩增、测序和系统发育分析。从宁夏苦马豆植株的叶和茎中共分离出4个真菌菌株,编号分别为SS_NXB1、SS_NXF1、SS_NXG2、SS_NXA1,因植株未表现出任何病害症状,故确定其为内生真菌。根据形态特征和ITS序列,确定前3株真菌为枝顶孢属(Acremonium)真菌。因菌株SS_NXA1未产生分生孢子及GenBank数据库中缺少同源菌株的分类信息而未确定其分类地位。  相似文献   

5.
为探索宁夏干旱荒漠区苦豆子(Sophora alopecuroides L.)内生真菌的多样性及区系组成,为苦豆子内生真菌的合理开发和利用提供理论依据。从宁夏白芨滩国家级自然保护区不同植被和土壤类型的6个样区采集苦豆子样品30份,采用组织匀浆法从植株的根、茎、叶和种子中分离内生真菌,根据培养性状、菌落、孢子等形态特征,结合rDNA-ITS序列分析对菌株进行鉴定;根据内生真菌的相对频率、物种多样性指数、丰富度指数和相似性系数分析其区系组成特点。结果表明,从30份样品中共分离得到内生真菌213株,分别属于19个属,其中镰刀菌属(Fusarium)、链格孢属(Alternaria)和茎点霉属(Phoma)为优势属。苦豆子内生真菌的分布具有一定的组织特异性,根部的内生真菌数量高于其他部位,茎部内生真菌多样性指数最高。植被类型中,沙生植被草原(样区Ⅴ)分离的内生真菌物种多样性指数最高,而荒漠草原(样区Ⅲ)最低。荒漠草原(样区Ⅰ)和沙生植被草原(样区Ⅴ)内生真菌群落有密切的相似性。可见,苦豆子蕴含丰富的内生真菌资源,其内生真菌具有很高的宿主特异性,且分布受生境影响。  相似文献   

6.
对宁夏黄花棘豆中产苦马豆素内生真菌进行分离和鉴定,为进一步研究疯草内生真菌合成苦马豆素的分子调控机理提供菌种和奠定基础。分离并纯化黄花棘豆植物组织中的真菌,采用气相色谱质谱联用法对各菌株菌丝中的苦马豆素进行分析,筛选能产生苦马豆素的内生真菌;紫外线照射和黑暗交替培养诱导内生真菌产孢;提取真菌DNA,扩增和测定真菌甘油醛-3-磷酸脱氢酶(glyceraldehyde-3-phosphate dehydrogenase,GPD)、内部转录间隔区(internal transcribed space,ITS)和线粒体核糖体小亚基(mitochondrial small subunit,mt SSU)基因序列,进行同源性比对和遗传进化分析,对其进行种属分类和命名。本研究从宁夏黄花棘豆中共分离到5株真菌,其中菌株NX-FEL001菌丝中含有苦马豆素,经诱导培养,该菌株能产生分生孢子及分生孢子梗等结构;序列同源性和系统发育分析结果表明,该菌株与Undifilum真菌的亲缘关系最近,根据形态结构和分子进化分析结果将菌株NX-FEL001鉴定为Undifilum oxytropis。  相似文献   

7.
为探明内生真菌在航天诱变材料黔草1号高羊茅中的分布情况和种类,利用光学显微镜进行镜检观察,对被侵染的航天诱变材料高羊茅展开内生真菌分离培养研究。结果发现,内生真菌的菌丝体普遍存在于试验材料的叶片、叶鞘、茎节间,内生真菌侵染率大小为:茎节叶鞘叶片,侵染差异在航诱材料与对照间(未经航天诱变的原始材料黔草1号)差异不显著(P0.05);对分离条件优化后,发现从试验材料中分离的内生真菌在pH 7.5,25℃黑暗培养的条件下以添加高羊茅煎煮液的马铃薯琼脂糖培养基PSA的培养效果最佳;在pH 7.5,25℃紫外光照/黑暗交替培养的条件下以添加高羊茅煎煮液的水琼脂培养基WSA的促孢效果最佳。  相似文献   

8.
青海高原的高寒特性决定其生境内禾草植物的多样性和特异性,然而有关青海高原地区禾草内生真菌的研究报道较少,本研究对青海高原地区48个样点内禾草植物内生真菌带菌率和内生真菌分离进行了研究.结果表明,14种禾本科植物内生真菌平均感染率低于50%,而12种植物内生真菌平均感染率大于50%.其中,缘毛鹅观草(Roegneria pendulina)、紫花针茅(Stipa purpurea)、醉马草(Achnatherum inebrians)、中华羊茅(Festuca sinensis)、硬质早熟禾(Poa sphondylodes)、微药羊茅(F.nitidula)、毛稃羊茅(F.kiritowii)和紫羊茅(F.rubra) 的平均带菌率均超过80%.同时,同属禾草植物内生真菌带菌率存在差异.禾草内生真菌分离率较低,其中紫花针茅、麦宾草(El ymus tangutorum)、光稃早熟禾(P.psiolepis)和异针茅(S.aliena)内生真菌在菌落和分生孢子形态上存在差异.可见,青海高原禾草内生真菌带菌率较高,同时不同禾草内生真菌可能存在多样性.  相似文献   

9.
我国3种疯草的内生真菌培养特性研究   总被引:1,自引:0,他引:1  
汪治刚  曹师  徐娜  徐杉  李彦忠 《草业学报》2017,26(10):158-169
家畜取食疯草出现中毒或死亡的原因为疯草体内含可产生苦马豆素的内生真菌。我国的多种疯草中的内生真菌为同一种,同种疯草中的内生真菌因随种子传播而孤立于单一寄主,可能出现与其他疯草内生真菌不同的生物学特性。为此在通过形态学和分子生物学鉴定菌种的基础上研究了我国最主要3种疯草小花棘豆、黄花棘豆和甘肃棘豆中内生真菌菌株(LYZ0091、LYZ0093和LYZ0109)在不同温度、pH值和培养基上的菌落生长速率和形态特性。结果表明,1)3个菌株的菌落停止生长的最高温度不同,LYZ0109无法在30 ℃下生长,但LYZ0091和LYZ0093仍可生长,最适宜生长温度分别为20、25和25 ℃,在5、10和15 ℃下4周时LYZ0091的生长速率显著(P<0.05)高于LYZ0109,在20和25 ℃下则显著(P<0.05)低于LYZ0109;2)在pH 4~11下3个菌株均可生长,菌株之间无显著(P>0.05)差异;在培养基NA、PCA、PDA、PSA和WHDA上LYZ0109多显著(P<0.05)大于LYZ0091;3)在温度、pH值和培养基处理中3个菌株均未产生分生孢子,LYZ0091和LYZ0109的菌丝扭曲状,厚垣孢子少,而LYZ093无扭曲状菌丝,大量成串的厚垣孢子由基质底部向上呈树状生长。  相似文献   

10.
为了研究昆虫寄生性真菌——棕色绿僵菌(Metarhizium brunneum)在家畜外寄生虫病防控中的应用,试验通过培养棕色绿僵菌获得分生孢子,稀释至浓度为1×104个/mL后统计其萌发率,在棕色绿僵菌分生孢子悬液对草原革蜱幼蜱和成蜱的杀灭作用试验中分别设置1个对照组和5个浓度的试验组,即将草原革蜱幼蜱和成蜱分别放入5个不同浓度的棕色绿僵菌分生孢子悬液及0.5%孢子洗脱液中停留5 s,在25℃条件下每日定时观察幼蜱和成蜱的虫体死亡情况及作用时间。结果表明:棕色绿僵菌培养至第3天时所得的分生孢子萌发率可达到90.40%;对草原革蜱幼蜱杀灭作用明显,用试验所得的最佳作用浓度1×106个/mL作用至第7天时草原革蜱幼蜱的死亡率高达100%,成蜱的死亡率为80.00%。说明昆虫寄生性真菌——棕色绿僵菌可以用于防治家畜外寄生虫病。  相似文献   

11.
为分析云南省虫媒病毒的种类与遗传特征,在云南省师宗县采集库蠓进行病毒的分离与鉴定;通过全长cDNA扩增与高通量测序技术获取病毒全基因组序列,进行序列比对与系统发生树构建。结果显示,从采集的库蠓样本中分离出1株可在C6/36细胞上引起细胞病变的毒株(YNSZ043),病毒基因组为分节段双链RNA,琼脂糖凝胶电泳呈"2-4-3"的带型特征;电镜观察可见直径为70~80 nm,呈"指环状",表面具有纤维突起的病毒粒子。全基因组测序结果显示,YNSZ043毒株为版纳病毒(Banna virus,BAV),基因组大小为20 683 bp,由Seg-1(3 762 bp)至Seg-12(861 bp)12个基因节段组成,与中国BAV毒株各基因节段的核苷酸序列相似性在64.8%~99.6%之间,氨基酸序列相似性在58.8%~100%之间,在系统发生树上YNSZ043毒株与中国分离的BAV聚为一簇,形成独立的中国进化支系。对决定BAV基因型的Seg-12分析结果显示,YNSZ043毒株属于A2基因型,该毒株的Seg-5/VP5与越南分离BAV毒株的核苷酸和氨基酸序列相似性高达97.1%和97.6%,表明该毒株的Seg-5基因节段很可能与越南毒株之间发生了基因重配。研究结果丰富了中国BAV的基因组序列,为开展云南省BAV的流行病学研究提供了参考。  相似文献   

12.
新鲜牛粪便中食线虫真菌的分离和鉴定   总被引:3,自引:1,他引:2  
从78个新鲜牛粪样中分离到4株食线虫真菌(nematophagous fungi),通过对其菌丝体、孢子、捕食器官等的形态学观察、测定,2株鉴定为节丛孢属(Arthrobotrys)的寡孢节丛孢菌(A.oligospora),1株为指状节丛孢菌(A.dactyloides),1株为圆锥状节丛孢菌(A.conoides)。  相似文献   

13.
A grazing experiment was conducted to assess the effects of wild-type endophyte-infected (E+) tall fescue consumption and elevated ambient temperatures on intravaginal temperatures, plasma lipid peroxidation, and glutathione redox of peripheral blood mononuclear cells. Angus heifers (n = 34) were allotted by BW to 4 blocks consisting of E+ and endophyte-free (E-) fescue pastures. Monthly, in June, July, and August, temperature loggers were fixed into blank controlled internal drug releasers and inserted into a subsample of heifers (n = 16) for 2 d. After 48 h, heifers were weighed, and blood (30 mL) was collected via jugular venipuncture. Peripheral blood mononuclear cells were isolated for analysis of glutathione peroxidase activity, glutathione reductase activity, and reduced:oxidized glutathione. Plasma malondialdehyde was evaluated as a marker of lipid peroxidation, and whole blood Se concentration was determined. Serum prolactin was assayed after the grazing period. Heifer ADG was greatest in August and least in July (P < 0.001). In August, heifers grazing E+ fescue exhibited greater (P < 0.05) afternoon intravaginal temperatures and temperature fluctuations than heifers grazing E- fescue. In July and August, all heifers had greater afternoon temperatures (P < 0.02) and less reduced:oxidized glutathione (P < 0.0001) than in June. Glutathione reductase activity of all heifers was greater in June (P = 0.03) than in July. Similarly, all heifers exhibited decreased glutathione peroxidase activity (P < 0.0008) in July, whereas whole blood Se was reduced (P < 0.0001) in July and August. No treatment or date effects were detected for malondialdehyde, but serum prolactin was reduced at the end of the grazing period (P = 0.008) in heifers stocked on E+ fescue. Using these markers, differences in oxidative stress were not detected between heifers consuming E+ fescue and those consuming E- fescue. Date effects indicating altered glutathione redox and enzyme activity may have been related to heat stress and nutritional limitations.  相似文献   

14.
The presence of porcine toxigenic E. coli (ETEC, VTEC) in 28 piggeries (5% of total) of the central and northeast region of Argentina was studied for a better understanding of the epidemiology of porcine strains. Samples were taken by rectal swabs from healthy piglets and from those with diarrhoea, in addition to their dams. Between 5-10 colonies were isolated from each one of 223 animals sampled from 1992 to 1997. By using specific primers each strain was screened by PCR for VT1, VT2all, VT2e, STIa, and LTI toxin genes. Only strains positive for any of the toxins mentioned above were screened for STb. Their O serogroups were determined by agglutination. All of the above enterotoxins and verocytotoxins were found in E. coli isolated from the animals. The STIa gene was detected in E. coli isolated from 27/127 piglets with diarrhoea, in comparison with LTI (4/127 pigs). No toxin gene was amplified from E. coli isolated from either healthy piglets or their dams. When strains isolated from 48 piglets without diarrhoea but showing delayed growth were analysed by PCR, their toxin profile was determined to be VT1 (1/48 piglets), VT2all (5/48), STIa (1/48), LTI (3/48) and VT2e (3/48). Serogroup O64 prevailed among ETEC; O138 prevailed for ETEC/VTEC strains. This is the first extensive study regarding porcine toxigenic E. coli in Argentina and constitutes an important database for the implementation of prevention measures.  相似文献   

15.
The cultural, morphological, biochemical, serological characteristics and antibiotic susceptibility of 25 Rhodococcus equi strains isolated from lungs and lung abscesses of pneumonic foals and 5 R. equi strains isolated from immuno-compromised human patients were examined. All R. equi strains showed common cultural, morphological and biochemical characteristics both with conventional tests and on the basis of their enzyme profile. The R. equi strains examined were resistant to penicillins with the exception of ampicillin, to sulphamethazine and several strains also to sulphamethoxazole-trimethoprim. All strains were susceptible to erythromycin and rifampicin. The strains isolated from humans showed somewhat higher rate of antibiotic resistance to penicillin, cefotaxime, kanamycin, streptomycin, lincomycin, and oxytetracycline. The overwhelming majority (96%) of the equine isolates belonged to serotype 1 in Prescott's serotyping system, while the human isolates could not be serotyped.  相似文献   

16.
A group of 48 crossbred steers (approx 250 kg each) were used to determine the effects of various diets and treatments on serum prolactin concentrations and rectal temperatures. The steers were randomly assigned to groups fed the following: (1) endophyte fungus-infested fescue hay; (2) ammoniated endophyte fungus-infested fescue hay; (3) endophyte fungus-infested fescue hay plus 1 g of thiabendazole (TBZ)/9.1 kg of body weight at 7-day intervals; (4) ammoniated endophyte fungus-infested fescue hay plus 1 g of TBZ/9.1 kg at 7-day intervals; (5) ground Bermuda grass hay; and (6) endophyte-fungus-free fescue hay. Blood samples for prolactin determinations and rectal temperatures were obtained biweekly beginning on week 3 and continuing through week 9. A significant correlation (P less than 0.05) between low prolactin concentrations and high rectal temperatures in cattle eating endophyte-infested fescue hay was determined; however, as the prolactin concentrations approached normal (control animal) concentrations, this relationship between serum prolactin and rectal temperatures was not observed. Two antifungal agents used in this experiment (thiabendazole and ammonia) appeared to have different effects on the variables measured. Thiabendazole had no significant effect on prolactin concentrations or rectal temperatures of cattle ingesting endophyte-infested fescue hay, whereas ammoniation of this hay induced significantly higher (P less than 0.05) prolactin concentrations and lower rectal temperatures than in steers receiving only endophyte-infested fescue hay. Therefore, ammonia may be valuable not only as an aid in determining the cause of the fescue problem but also as a practical solution to some of the fescue-related economic problems in the cattle industry. Also, prolactin concentrations may be valuable in diagnosing fescue-related problems.  相似文献   

17.
The aim of this study was to demonstrate the effect of auto-vaccine therapy on selected properties of Staphylococcus aureus strains, isolated from milk of cows with subclinical mastitis. The experiment was based on auto-vaccines which were prepared from S. aureus strains isolated from 16 cows. S. aureus strains isolated from cows on the 7th, 21st and 35th day following auto-vaccination, were analyzed phenotypically and genotypically. The isolated strains represented 17 biotypes all belonging to one clonal type. Increases of new biotypes of S. aureus were detected on the 35th day of therapy. Among 48 re-isolated strains, 18.75% (9/48) revealed single and 12.50% (6/48) multiple phenotypical changes. The present study demonstrated that during auto-vaccine therapy, S. aureus strains can change phenotypically, pointing out the necessity for using precise diagnostic methods, that would make possible a better assessment of the used therapy.  相似文献   

18.
宋辉  南志标  田沛 《草业学报》2015,24(9):89-95
内生真菌与植物所形成的共生体普遍存在于冷季禾草中,共生体有利于提高宿主植物的生物和非生物抗性。虽然目前研究者们已对禾草内生真菌做了大量的研究,但是尚未发现对分布于中国西北地区披碱草属植物无性世代Neotyphodium内生真菌培养特征的研究。本研究从中国西北地区分离出15株来自不同地理种群披碱草属植物无性世代Neotyphodium内生真菌。通过分离培养,测定了这15株无性世代Neotyphodium内生真菌的菌落生长速度,结果显示,这15株无性世代Neotyphodium内生真菌菌落的平均生长速度为(0.41~0.85) mm/d,且生长速度与海拔具有相关性。分布在海拔低于3000 m的无性世代Neotyphodium内生真菌的菌落生长速度比生长于海拔高于3000 m菌落的生长速度快。观察菌落发现,分布在海拔高于3000 m的菌落正面特征较为一致,而分布在海拔低于3000 m的菌落正面特征出现不同。另外,通过多基因联合建树发现,分布在海拔高于3000 m的无性世代Neotyphodium内生真菌聚在一个分支,而分布在海拔低于3000 m的无性世代Neotyphodium内生真菌呈现星状分布。由于目前尚未对中国西北地区披碱草属植物无性世代Neotyphodium内生真菌命名。因此,希望本研究结果能为该菌的命名提供理论依据。  相似文献   

19.
动物蓝舌病毒L2基因克隆及序列分析比较   总被引:2,自引:0,他引:2  
对 15株中国动物蓝舌病毒血清 1型 (BTV- 1)野毒株及 1株弱毒疫苗株 L 2基因进行了克隆和测序 ,并进行了系统进化分析。中国 BTV- 1型毒株分为 2个组 :分离年代最早的 Y86 3毒株 (1979年 )为 组 ;1996年以后分离的 14株毒株为 组。 2组之间的同源性是 91.6 %。 组又分为 4个谱系 ,疫苗株 Vac F45与原始株 V6 5 8分别属于 组的第 1和第 3谱系 ,核苷酸同源性 98.6 %。由此表明 ,中国 BTV- 1型毒株在自然进化及鸡胚传代驯化过程中 ,L2基因发生了遗传变异。  相似文献   

20.
从青贮饲料中分离鉴定了39株乳酸菌,并研究了这些乳酸菌的特性。结果表明:分离菌株都是革兰氏阳性、过氧化氢酶阴性、兼性厌氧菌,根据其形态学和生理生化特性,可分为7组(A-G),A组、B组为片球菌属(Pediococcus),C组、D组和E组为肠球菌属(Enterococcus),F组和G组属于乳杆菌属(Lactobacillus)。  相似文献   

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