单克隆抗体对小苍兰褪绿条纹花叶病毒及其决定簇的作用特性

采用杂交瘤技术,得到抗小苍兰褪绿色条纹花叶病毒（FCSMV）六个单克隆抗体P3、4A4,6B、,7A6,7D1和7DS。ELISA效价分别为1：1062－1：10^6。7A6攻7D1属于IgG1亚类,7D8和4A4分属IgG2a和IgG2b亚类,P3和6B1属于IgG2亚类。电镜观察单克隆抗体与病毒形成的复合物发现7D1能使病毒粒子发生严重裂断。6株单克隆抗体中P3,4A4、6B1对应于病毒外壳蛋白上的顺序决定簇。纯化的病毒外壳蛋白经酶裂解和改进的盘状电泳分离,得到八段与多克隆抗体反应的收集液。用顺序决定族的单克隆抗体检出P3对应的肽段22和4A4对应的肽段78。本文还讨论了7D1使病毒粒子发生断裂的原因和与病毒粒子表面结构的关系以及分离抗原决定族所在片段的意义。     

侵染花生的辣椒褪绿病毒S RNA全序列分析

番茄斑萎病毒属(Tospovirus)是布尼亚病毒科(Bunyaviridae)中植物病毒组成的一个属, 病毒粒子为球状,直径80～110nm,粒体外层由一层脂质包裹.基因组属于负单链RNA,由三个片段组成,分别被称为L RNA、M RNA、和S RNA.L RNA为负链、含单个开放阅读框架(ORF),M RNA和S RNA均为双义RNA, 有2个ORF,反向相连.     

侵染花生的辣椒褪绿病毒SRNA全序列分析

番茄斑萎病毒属(Tospovirus)是布尼亚病毒科(Bunyaviridae)中植物病毒组成的一个属,病毒粒子为球状,直径80~110nm,粒体外层由一层脂质包裹。基因组属于负单链RNA,由三个片段组成,分别被称为L RNA、M RNA、和S RNA。L RNA为负链、含单个开放阅读框架(ORF),M RNA和S RNA均为双义R     

我国菊花褪绿斑驳类病毒鉴定

从我国广州地区发现的带有褪绿斑驳症状的“广白”菊花病株中分离到一种小分子核酸,经鉴定其性质与国外报道的菊花褪绿斑驳类病毒(Chrysanthemum Cholrotic Mottle Viroid,ChCMV)的性质完全一致。病株用酚提取后的粗核酸和制备的纯核酸经正反双向或垂直双向聚丙烯酰胺凝胶电泳分析表明为具有单链环状的RNA分子,其分子量与菊花矮化类病毒(Chrysanthemum Stunt Viroid, CSV)的相似。根据病株症状、寄主范围以及分段自我杂交等试验分析,证明这种小分子核酸是类病毒——chcMV,而不是CSV。在检测过程中,我们改进了提取和鉴定类病毒的方法,并建立了可以区分不同种类病毒的分段自我杂交技术。     

番茄褪绿病毒对刀角瓢虫捕食烟粉虱的影响

在实验室条件下研究了不同温度下刀角瓢虫Serangium japonicum对健康和感染番茄褪绿病毒(Tomato chlorosis virus,ToCV)番茄上的烟粉虱Bemisia tabaci的捕食效果.结果表明,无论在健康番茄还是感染ToCV番茄上,刀角瓢虫均可以取食烟粉虱的卵和若虫,对卵的捕食量最大,且对烟...     

杏褪绿卷叶病研究现状及展望

杏褪绿卷叶病(ACLR)是由ESFY植原体引发的一类昆虫媒传病害,在欧洲许多国家已将其列为检疫对象。近年来在我国新疆部分杏园首次发现类似欧洲ACLR的发病植株。简要介绍杏褪绿卷叶病的田间发病症状,疏理了国内外关于引起杏褪绿卷叶病的ESFY植原体的流行病学和病原检测以及防治方法的研究进展,特别对分子检测技术进行全面综述。另外,分析当前研究中尚存在的问题,并对今后的研究方向进行展望。     

烟粉虱MEAM1和MED成虫在辣椒上传播番茄褪绿病毒的特性

【目的】分析与比较烟粉虱Bemisia tabaci MEAM1和MED成虫在辣椒Capsicum annuum植株上对番茄褪绿病毒(tomato chlorosis virus, ToCV)的传播差异,解析媒介昆虫-病毒-寄主植物间的互作关系,为田间辣椒上ToCV防治提供参考。【方法】利用烟粉虱MEAM1和MED成虫及ToCV cDNA侵染性克隆,对烟粉虱MEAM1和MED成虫在辣椒植株上获取和传播ToCV的能力进行比较,同时比较烟粉虱MEAM1和MED成虫在健康和携带ToCV辣椒植株上的取食偏好。【结果】在感染ToCV的辣椒植株上接虫后96 h内,烟粉虱MEAM1和MED成虫体内病毒积累量逐渐增加,MED成虫获毒速度和病毒积累量均大于MEAM1成虫的,MED成虫获毒量是MEAM1成虫的1.74倍;MED成虫在辣椒植株上的传毒量明显高于MEAM1成虫,传毒量平均相差9倍,且MED成虫传毒率比MEAM1成虫高29%。MEAM1和MED成虫对健康和感染ToCV的辣椒植株的取食偏好相似,都明显倾向于取食被ToCV侵染的辣椒植株。【结论】烟粉虱MED成虫在辣椒植株上获取、传播ToCV的能力均...     

苹果褪绿叶斑病毒和茎沟病毒的鉴定提纯和酶联法检测

苹果褪绿叶斑病毒(CLSV)和茎沟病毒(scv)是感染苹果和其它果树的两种重要潜隐 病毒。根据生物学特性和血清学反应,确定W-55分离物属于CLSV,W-29分离物属于SGV。采用皂土澄清,PEG 6000沉淀及超离心等步骤,得到部分提纯的病毒。经电镜观察,W-55和 W-29病毒颗粒长度分别为800nm和650nm。用CLSV W-55分离物制备的兔抗血清效价为1／3200,用SGV PV一71(ATCG)分离物制备小鼠腹水抗体效价为1／800,均用间接ELISA法测定。对感染CLSV和SGV的昆诺藜、苹果叶和花瓣检测结果表明,P／N值具有显著性。本工作为苹果无毒苗木的生产提供了快速、灵敏的病毒检测手段。     

番茄褪绿病毒TaqMan荧光定量PCR快速检测方法的建立与应用

【目的】本研究旨在建立TaqMan实时荧光定量PCR(TaqMan RT-qPCR)技术,快速检测单头烟粉虱Bemisia tabaci体内的番茄褪绿病毒(tomato chlorosis virus, ToCV)。【方法】根据ToCV外壳蛋白保守序列设计了1对特异性引物和1条TaqMan探针,建立了TaqMan RT-qPCR方法;与常规PCR检测进行比较,检测该方法的灵敏度与特异性;并应用该方法对单头烟粉虱成虫体内ToCV进行了快速检测。【结果】本研究构建的TaqMan RT-qPCR检测ToCV的标准曲线,其循环阈值(Ct值)与模板浓度具有良好的线性关系,扩增效率为98%。该方法对ToCV的最低检测浓度为8.3×10 copies/μL,灵敏度是常规RT-PCR的1 000倍。该方法与田间番茄两种重要病毒番茄黄化曲叶病毒(tomato yellow leaf curl virus, TYLCV)和番茄斑萎病毒(tomato spotted wilt virus, TSWV)检测无交叉反应。单头烟粉虱成虫ToCV检测结果表明,温室内ToCV侵染植株上烟粉虱携毒率为100%,田间烟粉...     

番茄褪绿病毒对Q型烟粉虱重要生物学参数及保护酶和解毒酶活力的影响

为明确番茄褪绿病毒(ToCV)对其传毒介体烟粉虱Bemisia tabaci主要生物学特性的影响,本文研究了携带ToCV的Q型烟粉虱在非病毒寄主植物棉花Gossypium spp上的生物学指标,并测定了带毒和无毒烟粉虱主要保护酶和解毒酶活性.结果 表明,在棉花植株上,带毒烟粉虱在发育历期、产卵量、成虫寿命方面与无毒烟粉虱无显著差异,但雌虫体长明显短于无毒烟粉虱.相对于无毒烟粉虱,带毒烟粉虱体内过氧化氢酶(catalase,CAT)活性明显提高,是无毒烟粉虱的3.36倍(P ＜0.001),超氧化物歧化酶(superoxide dismutase,SOD)和过氧化物酶(peroxidase,POD)活性无显著差异.解毒酶中,带毒烟粉虱羧酸酯酶(carboxylesterase,CarE)活性明显下降,是无毒烟粉虱活性的54％,谷胱甘肽S转移酶(glutathione-s-transferase,GST)和乙酰胆碱酯酶(acetylcholin esterase,ACHE)活性无显著差异.     

Properties of Johnsongrass Chlorotic Stripe Mosaic Virus

Physicochemical, biological, and cytopathological properties of Johnsongrass chlorotic stripe mosaic virus (JCSMV) found in Iran were investigated. Virus particles were polyhedral, showed a knobbed surface structure, were c. 30 nm in diameter and had a buoyant density of 1. 359 g/ml in cesium chloride. Virions contained one major protein with a molecular weight of 41 kd and a single species of ssRNA with a molecular weight of 1. 43 × 10⁶ d. Acid hydrolysis of the virus followed by thin-layer electrophoresis gave the following molar percentages of the bases: A: 23. 5, G: 27. 5, C: 26 and U: 23. Separation of nucleotides of the viral RNA using alkaline hydrolysis was not successful. Mechanical inoculation of freshly purified virus or isolated RNA failed to infect Johnsongrass or maize plants. The virus was readily detected by ELISA in seeds from infected plants and young seedlings raised from such seeds, but not in later stages of growth. Ultrathin sections of infected cells showed high concentrations of virus particles in the cytoplasm and vacuoles. Virus-like particles also occurred in the stroma of chloroplasts and mitochondria. Cisternae of endoplasmic reticulum (ER) were often extremely inflated and filled by a fine fibrillar material. Small membrane-associated vesicles were frequently found in ER elements and occurred also in the permuclear space. Based on particle structure, properties of the nucleic acid, molecular weight of the coat protein and cytopathology, the virus resembles carmoviruses. However, lack of mechanical transmissibility is not known for any virus classified with this group. No serological reaction was detected with a total of 30 antisera to carmoviruses and other isometric viruses.     

Engineering Tobacco Mosaic Virus and Its Virus-Like-Particles for Synthesis of Biotemplated Nanomaterials

Biomolecules are increasingly attractive templates for the synthesis of functional nanomaterials. Chief among them is the plant tobacco mosaic virus (TMV) due to its high aspect ratio, narrow size distribution, diverse biochemical functionalities presented on the surface, and compatibility with a number of chemical conjugations. These properties are also easily manipulated by genetic modification to enable the synthesis of a range of metallic and non-metallic nanomaterials for diverse applications. This article reviews the characteristics of TMV and related viruses, and their virus-like particle (VLP) derivatives, and how these may be manipulated to extend their use and function. A focus of recent efforts has been on greater understanding and control of the self-assembly processes that drive biotemplate formation. How these features have been exploited in engineering applications such as, sensing, catalysis, and energy storage are briefly outlined. While control of VLP surface features is well-established, fewer tools exist to control VLP self-assembly, which limits efforts to control template uniformity and synthesis of certain templated nanomaterials. However, emerging advances in synthetic biology, machine learning, and other fields promise to accelerate efforts to control template uniformity and nanomaterial synthesis enabling more widescale industrial use of VLP-based biotemplates.     

侵染番茄的番茄花叶病毒的研究

从种传番茄苗中获得一病毒分离物Ｔｏ－Ｓｌ,人工摩擦接种７科２４种植物,Ｔｏ～Ｓｌ能侵染４科１５种植物,在番茄上产生花叶,在白肋烟上为局部枯斑。Ｔｏ－Ｓｌ的钝化温度为８５～９０℃,稀释限点为１０￣（－６）～１０￣（－７）．体外保毒期在一个月以上。病毒粒体杆状,长度主要分布于２８１～３００ｎｍ之间,平均长度２８８ｎｍ。病毒衣壳蛋白亚基只有一条多肽链,分子量为２１ｋＤａ。ｄｓＲＮＡ分析测得其基因组长度为６．４ｋｂ。琼胎糖双扩散和胶内交叉吸附试验证明,Ｔｏ－Ｓｌ与ＴＭＶ有血清关系,但有一定的差异,病毒粒体电泳分析也表明Ｔｏ－Ｓｌ与ＴＭＶ粒体有差异。在交叉保护试验中,ＴＭＶ和Ｔｏ－Ｓｌ之间均无保护作用。根据以上试验结果Ｔｏ－Ｓｌ被鉴定为番茄花叶病毒。这是我国首次系统报道番茄上番茄花叶病毒的侵染。     

Volatile Compounds in the Flowers of Freesia Parental Species and Hybrids

For centuries, freesia has been one of the most important crops in the floriculture industry. Here, aqua-space samples collected from entire flowers of diploid Freesia refracta, three tetrapioid freesia cuitivars, and interspecific hybrids of three tetraploid freesia cultivars were analyzed using gas chromatograph coupled with mass selective detector, in all, 75 different compounds were identified. The compounds were mainly terpenes, hydrocarbons, alcohols, fatty acid esters and aromatic class compounds. Among these, iinalool was detected from all the sweet-scented flowers except for scentless white tetraploid F. hybrida. Stable inheritance of linalool between F. hybrida and their Ft progeny was observed. Based on the present analyses, the relationship between the aroma of freesia and iinalooi was discussed.     

Monoclonal Antibody-Based Serological Detection of Rice Stripe Mosaic Virus Infection in Rice Plants or Leafhoppers

Rice stripe mosaic virus(RSMV) is a rhabdovirus recently found in southern part of China and can cause severe reduction in rice production. To establish serological methods for RSMV epidemiological studies and to establish a control strategy for this virus, we first purified RSMV virions from infected rice plants and then used them as an immunogen to produce four RSMV-specific monoclonal antibodies(MAbs)(i.e.,1D4, 4A8, 8E4 and 11F11). With these MAbs, we have developed a highly specific and sensitive antigen-coated plate enzyme-linked immunosorbent assay(ACP-ELISA), a Dot-ELISA and a Tissue print-ELISA for rapid detections of RSMV infection in rice plants or in leafhoppers. Our results showed that RSMV can be readily detected in RSMV-infected rice plant tissue crude extracts diluted at 1:20,971,520(w/v, g/m L)through ACP-ELISA or diluted at 1:327,680(w/v, g/m L) through Dot-ELISA. Both ACP-ELISA and Dot-ELISA can also be used to detect RSMV infection in individual RSMV viruliferous leafhopper(Recilia dorsalis) homogenate diluted at 1:307,200 and 1:163,840(individual leafhopper/l L), respectively. Detection of RSMV infection in field-collected rice samples or in RSMV viruliferous leafhoppers indicated that the three serological methods can produce same results with that produced by RT-PCR(19 of the 33 rice samples and 5 of the 16 leafhoppers were RSMV-positive). We consider that the four MAbs produced in this study are very specific and sensitive, and the three new serological methods are very useful for detections of RSMV infection in rice plants or in leafhoppers and the establishment of the disease control strategies.     

应用免疫电镜技术快速检测菜豆黄花叶病毒、马铃薯M病毒和燕麦花叶病毒

应用免疫吸附电流技术(ISEM)可有效地检测腐汁液中的菜豆黄花叶病毒(BYMV)、马铃薯M病毒(PVM)和燕麦花叶病毒(OMV)。BYMV,PVM和OMV三种抗血清的适宜工作浓度和对铜网的适宜包被时间均为1000倍和1小时,对同源病毒的适宜捕获时间分别为4℃下2、2和8小时。PVM和OMV的病汁液检测灵敏度均为稀释4000倍,而BYMV病汁液稀释16000倍时还能检测到少量病毒料子。ISEM捕获法和修饰法的结果表明,这三种病毒之间无血清学交叉反应。     

6个香雪兰品种的染色体核型及聚类分析

以香雪兰（Freesia hybrida）6个品种的根尖为研究材料进行核型分析以揭示其核型特征,进一步通过聚类分析品种间的相似性并探讨可能的亲缘关系。结果表明：香雪兰6个品种均为四倍体,染色体数目都为44。6个品种的核型公式不同：‘Gold River’为2n=4x=44=33m+11sm,‘White River’为2n=4x=44=1M+34m+9sm,‘Castor’为2n=4x=44=35m+9sm,‘上农红台阁’为2n=4x=44=32m+12sm,‘Pink Passion’为2n=4x=44=38m+6sm,‘Red River’为2n=4x=44=37m+7sm。核型参数因品种而异,其中,‘Castor’在相对长度范围和臂比范围上均为最广,‘上农红台阁’核不对称系数最大。6个品种核型不对称系数为57.00%~60.07%,核型类型均为2B,进化程度中等,处于由对称至不对称的过渡阶段,其核型进化的趋势大致为：‘Pink Passion’→‘Red River’→‘Gold River’→‘White River’→‘Castor’→‘上农红台阁’。聚类结果显示,遗传距离为15时可将所有品种分为两类,第一类包含‘Gold River’‘上农红台阁’和‘Castor’; 第二类包含‘Pink Passion’‘Red River’和‘White River’,其核型参数的差异可作为香雪兰品种分类和鉴定的辅助依据,为其细胞学研究和育种实践提供理论参考。     

番茄花叶病毒单克隆抗体的制备及检测应用

用番茄花叶病毒(ToMV)免疫的BAL B/c鼠脾细胞与SP2/0鼠骨髓瘤细胞融合,经筛选克隆,获得4株能稳定传代并分泌抗ToMV单克隆抗体(Mab)的杂交瘤细胞,其中2株能同时检测ToMV和烟草花叶病毒(TMV),各单克隆抗体腹水ELLSA效价在1∶32 000～1∶1 024 000之间。经TASELISA测定,4株单克隆抗体检测病汁液的稀释度均能达到1∶2 000倍以上。4株单克隆抗体与其他病毒无交叉反应。Westernblot分析表明,其中两株与ToMV176kD的外壳蛋白亚基有特异反应,而另两株无反应,推测它们是针对构象决定簇的抗体。     

山东烟台小麦土传病毒病由小麦黄花叶病毒和土传小麦花叶病毒相关病毒复合侵染所致

在山东省烟台地区的小麦上发生一种由土壤中禾谷多粘菌Polymyxa graminis传播的病毒病,感病小麦植株表现矮化褪绿和花叶症状.我们于1997年4月从病区采集感病小麦植株,进行了病毒种类鉴定.直接电镜观察发现有二种病毒粒子,一种粒子呈棒状,占大多数,其长度约为300nm和150nm; 另一种粒子呈线状,数量较少,长度为500nm～700nm.免疫电镜结果表明,棒状病毒粒子仅与土传小麦花叶病毒(soil-borne wheat mosaic virus, SBWMV)抗血清反应,而不与小麦黄花叶病毒(wheat yellow mosaic virus,WYMV)抗血清和小麦梭条斑花叶病毒(wheat spindle streat mosaic virus,WSSMV)抗血清反应;反之,线状病毒仅与WYMV、WSSMV抗血清反应,而不与SBWMV抗血清反应.用WYMV和SBWMV两种抗血清同时进行修饰时,线状病毒粒子和棒状病毒粒子均发生反应.