Characterization of an isolate of beet mosaic virus from South Kazakhstan

A filamentous virus isolated from a sugar-beet plant showing systemic mosaic collected in South Kazakhstan was identified as an isolate of beet mosaic virus (BMV-K). BMV-K was transmitted by the green peach aphid Myzus persicae in a non-persistent manner, and by sap inoculation to 11 out of 19 species from seven families tested. The virus could not be transmitted to Nicotiana tabacum, N. debneyi, N. glutinosa and N. clevelandii, cither mechanically or with M. persicae. The thermal inactivation point of BMV-K in sugar-beet sap was 55-60 C, dilution end point 1:1000 and longevity in vitro 2 days at 20 C. A purification procedure produced 1-5-3 mg of purified virus from 100 g of infected Stellaria media plants. Purified virus contained a single protein species of molecular weight 34 700 Da. In ELISA tests, BMV-K reacted positively with BMV-specifc antisera obtained from Japan. Germany and Portugal. By competitive DAS- ELISA, the virus isolate was shown to be closely serologically related to all the three isolates of BMV, and very distantly related to bean yellow mosaic and soy bean mosaic viruses.     

烟草坏死病毒A大豆分离物的分子鉴定

 对曹琦和濮祖芹早期分离到的烟草坏死病毒大豆分离物的生物学、血清学和外壳蛋白的序列进行了进一步研究。该分离物能侵染8科29种植物, 除系统侵染大豆和本生烟外, 其余寄主均为局部侵染。电镜下病毒粒子呈球状, 直径约28nm。基因组为单组分RNA, 大小约为3.7 kb, 具有2条亚基因组, 分别约为1.6 kb和1.3 kb。外壳蛋白亚基的分子量约为30 kDa。血清学试验表明, 该分离物与TNV柳树分离物的抗血清呈特异反应, 与同属坏死病毒属(Necrovirus)的烟草坏死病毒D(TNV-D)和甜菜黑色焦枯病毒(BBSV)无血清学关系。利用简并引物通过RT-PCR克隆了该分离物的外壳蛋白基因。序列分析表明, 该分离物与烟草坏死病毒A(TNV-A)、TNV-D和TNV-D^H的外壳蛋白分别具有88.77%、45.13%和45.49%的氨基酸序列一致性。因此, 该大豆分离物属于TNV-A的一个新株系, 命名为TNV-A^C。     

Partial purification of a virus associated with a Spanish isolate of citrus ringspot

A citrus ringspot isolate from Star Ruby grapefruit (RS-SR) was mechanically transmitted to Chenopodium quinoa. RS-SR was partially purified by differential centrifugation, fractionation in a sucrose gradient, and agarose gel electrophoresis of selected fractions. Infectivity of concentrated extracts on C. quinoa was lost in individual fractions of the gradient, but it was recovered by combining a top and a bottom component. Both components contained a 48-kDa protein not found in similar preparations from healthy plants. After further purification the 48 kDa protein was detected at the top edge of the agarose gel. In the initial experiments a 38-kDa protein was found in the same fractions that later contained the 48-kDa protein. An antiserum obtained to the 38 kDa protein reacted in Western blots with both the 38- and the 48-kDa proteins, whereas another antiserum raised to the Florida isolate CRSV-4 (also containing a 48-kDa protein) did not react with the 38-kDa protein, indicating that the latter was probably a degradation product of the 48-kDa protein. Filamentous flexous particles were observed by serologically specific electron microscopy in crude extracts from RS-SR-infected C. quinoa plants. These results indicate that RS-SR is associated with a two-component virus similar to those associated with several psorosis and ringspot isolates, and serologically related to CRSV-4.     

Characterization and vector relation of a serologically distinct isolate of tobacco rattle tobravirus (TRV) transmitted byTrichodorus similis in northern Greece

A virus isolated from diseased tobacco plants growing in Macedonia, northern Greece, had host range and physico-chemical properties typical of a tobravirus. Although it was serologically unrelated to any of the ten tobravirus isolates tested, it reacted in spot hybridization tests with a probe derived from RNA-1 of tobacco rattle virus (TRV) strain SYM. Therefore, the isolate probably represents a previously undescribed serotype of TRV. Male, female and juvenileTrichodorus similis nematodes recovered from the rhizosphere of the diseased tobacco plants transmitted TRV in each of three laboratory experiments. In two of these experiments 50% and 54%, respectively, of the nematodes transmitted virus toPetunia hybrida bait plants, whereas only 18% transmitted virus toNicotiana tabacum plants in a third test. Ultrathin sections of the feeding apparatus of individual nematodes, which had transmitted virus, were examined by electron microscopy. Virus particles were observed, retained as a monolayer in the apices of the oesophageal lumen and as a group of particles within a matrix in the open part of the lumen.     

引起辣椒花叶、枯顶的一个病毒分离物的鉴定

 从河北望都辣椒地分离到一株辣椒病毒分离物.测定表明能侵染茄科、苋科、豆科、菊科、藜科的25种植物,不能侵染葫芦科和十字花科等13种植物.桃蚜（Myzus persicae）非持久性传毒;钝化温度65—70℃,稀释限点10^-4—10^-5体外保毒期6天（20—22℃）;病毒颗粒呈球形,直径约25nm;ISEM测定它与蚕豆萎蔫病毒（BBWV）关系密切;电镜下观察到BBWV所特有的长管状内含体和布纹状结晶体,认为该分离物是BBWV.但寄主范围和寄主反应与文献报道的BBWV各分离物有所不同,可能是另一毒株。     

The partial characterization of melon vein-banding mosaic virus, a newly recognized virus infecting cucurbits in Taiwan

A potyvirus (designated as no. 656) causing mosaic or vein-banding symptoms on melons was isolated and characterized. The virus was mechanically transmitted to 14 herbaceous plant species, and induced mosaic symptoms in most cucurbitaceous plants. Aphis gossypii transmitted the virus non-persistently, and flexuous filamentous virus particles c. 755 nm in length were consistently observed in extracts of the infected pumpkin leaf tissues. Pinwheel and tubular inclusions were observed in the cytoplasm of infected zucchini leaf tissues. The virus was purified from infected pumpkin leaves by isopycnic centrifugation (Cs₂SO₄). The molecular weights of purified capsid and cylindrical inclusion proteins as estimated by SDS-PAGE were 32·5 and 73 KD_a, respectively. In SDS-immunodiffusion tests, antiserum to virus particles from isolate 656 was serologically unrelated to ZYMV, WMV-2, PRV-W and a type W variant of PRV, but antiserum to its cylindrical inclusion protein did produce spur precipitin bands between homologous and WMV-2 antigen wells. However, neither WMV-2 virus particle nor cylindrical inclusion antisera reacted with this virus. Furthermore, this virus was not serologically related to BCMV, BYMV, CYVV, SMV, WMV-M or ZYFV. Based on test results and symptomatology, this virus appears to be a new potyvirus, for which the name melon vein-banding mosaic virus (MVbMV) is proposed.     

中国水仙上一种球形病毒的研究Ⅰ.病毒分离及其生物学特性

 从水仙黄条病株上分离到一种球形病毒,直径约21nm,汁液摩擦接种35科(属)55(品)种植物,可侵染其中5科15(品)种,主要局限于豆种(Leguminosae)、藜科(Chenopodiaceae)以及茄科(Solanaceae)、苋科(Amaranthaceae)与番杏科(Aizoaceae)的个别种。在多数寄主上诱导局部坏死,豆科寄主上还伴随叶脉坏死症状产生。其中,昆诺阿藜(Chenopodium quinoa)、苋色藜(C.amaranticolor)、千日红(Gompherena globosa)、番杏(Tetragonia expansa)、大豆(Glycine max)、豇豆(Vigna sinensis)、克利夫兰烟(Nicotiana clevelandii)可作为诊断寄主;千日红、番杏与三叶草(Trifolium sibth)可作为毒源保存寄主。#br#在昆诺阿藜叶片汁液中,该病毒分离物的体外存活期在60天以上(25℃),热钝化点80-85℃(10分钟),稀释限点10^-5-10^-6。回接水仙健株,诱导产生系统褪绿条纹症状,不诱导水仙叶片产生具有特殊结构的内含体。     

Separation and interference of strains from a citrus tristeza virus isolate evidenced by biological activity and double-stranded RNA (dsRNA) analysis

Separation of strains of citrus tristeza virus (CTV), differentiated by their double-stranded RNA (dsRNA) profiles, was obtained by graft-inoculating citron plants from a Mexican lime that had been recently aphid- or graft-inoculated with a mild CTV isolate (T-385). Up to 24 sub-isolates with differing dsRNA profiles were obtained from the aphid-inoculated lime. Some of these sub-isolates induced stronger symptoms in several citrus species than the original T-385 isolate. One sub-isolate, T-385-33, was mild in Mexican lime, but induced stem pitting on sweet orange. Inoculation of this isolate on Mexican lime, sour orange and Eureka lemon induced mild or no symptoms when inoculum was taken from citron, but very severe symptoms when the inoculum was from sweet orange. Mexican lime and sweet orange plants co-inoculated with T-385-33 from sweet orange in combination with the other 23 sub-isolates showed mild symptoms. The results obtained suggest that there is natural cross-protection among sub-isolates in the original T-385 isolate.     

一种蚜虫持久性传病的长豇豆黄化型病毒

 1983年6月,在南京郊区的长豇豆上采到1株表现植株矮缩症状的C-7病毒分离物。接种试验证明,它不能摩擦接种传病,但可以由豆蚜(Aphis craccivora)、棉蚜(A.gossypii)和桃蚜(Myzus persicae)以持久性方式传病。寄主范围测定的结果表明:分离物可以侵染长豇豆、豇豆、蚕豆、大豆、菜豆、豌豆、赤豆、利马豆、苜蓿、红三叶、地三叶、绛三叶、葫芦巴,紫云英和苕子等15种豆科植物和曼陀罗1种茄科植物。这些植物大都出现植株矮化,叶片扭曲,卷缩或僵缩,不能开花结实等症状。豆蚜的传病性状中,获毒饲育的最短传病时间为3小时,接毒饲育最短传病时间为10分钟,循回期是24小时左右。但是,传病率最高的获毒饲育时间是2~3天,接毒饲育时间在1天以上。接种1头蚜虫就具有传病能力,5头蚜虫能达到100%的传病率。蚜虫可以终身传毒,蜕皮不影响其传毒力,但传毒有间歇性。根据它的基本性状,病毒C-7分离物是一种豆科植物的黄化型病毒,可能是属于大麦黄矮病毒组(Luteovirus Group)的成员。     

Some properties of alfalfa mosaic virus isolated from Buddleia davidii in the UK

Alfalfa mosaic virus (AMV) and cucumber mosaic virus (CMV) were isolated from clones of Buddleia davidii collected at Long Ashton Research Station for assessment of horticultural characteristics. The AMV isolate investigated in this study (AMV-B) infected 18 species and cultivars of herbaceous test plants. The in vitro properties of AMV-B were determined. The virus was purified by permeation chromatography on controlled-pore glass beads and an antiserum prepared. AMV-B appeared to be serologically indistinguishable from AMV isolate 15/64. The sizes and shapes of the virus particles were those of classical AMV components. The capsid protein was a single polypeptide of Mr 24 375. Buddleia seedlings inoculated with AMV-B showed only mild chlorosis and slight distortion of leaves.     

Filamentous flexous particles and serologically related proteins of variable size associated with citrus psorosis and ringspot diseases

Filamentous flexous partic les of unusual morphology, previously associated with several ringspot isolates, were detected also in psorosis A and psorosis B isolates by serologically specific electron microscopy using an antiserum to citrus ringspot. Upon partial purification of six ringspot, six psorosis A, and three psorosis B isolates, a specific protein of 47 kDa was detected in most cases, but two isolates (one psorosis A and one ringspot) had a 46 and a 48 kDa-protein, respectively. These differences in molecular masses were observed when purification was done from different host species or from plants co-inoculated with two isolates differing by their protein size. The three types of protein were serologically related in Western blots. Our results indicate that a common virus with different strains may be involved in psorosis A, psorosis B, and ringspot diseases.     

南京地区豇豆蚜传花叶病毒的鉴定

 1982年5月,从南京郊区豇豆花叶病植株上分离到1株病毒分离物C-1,接种试验的结果证明,它可以侵染12种豆科和藜科植物。它在豇豆上引起系统花叶、叶片卷曲、明脉和畸形等症状。它在苋色藜、昆诺藜和蚕豆上表现为局部病斑。体外抗性测定,失毒温度55~60℃,稀释限点10^-3~10^-4,体外存活期1~2天。病毒极易摩擦接种传病。桃蚜、棉蚜和豆蚜都能传染这种病毒。人工接种的豇豆病株,在花器的各个部分、幼嫩的豆荚组织和末成熟的种子内都带有病毒。病株上采收的种子传毒率可达8.1%。病毒存在于种子的胚和子叶内,种皮内没有测到病毒。病毒粒体线条状,长700~750纤米。病株叶片表皮细胞内有纺锤状的内含体。免疫电镜和SDS~双扩散法测定,病毒分离物C-1与豇豆蚜传花叶病毒(CAMV)的抗血清呈阳性反应。根据以上这些性状,病毒分离物C-1可鉴定为属于马铃薯Y病毒组中的豇豆蚜传花叶病毒。用微量沉淀法测定,病毒粗提纯液制备的抗血清的效价为1:512。SDS-双扩散法测定,南京地区严重发生的豇豆花叶病中,85~86%是由豇豆蚜传花叶病毒引起的。从福建、山东、辽宁等省采集的样本中,也证实这种病毒在豇豆上普遍发生。     

从新疆哈密瓜分离到的烟草坏死病毒及其理化性质的研究

 从自然感病的哈密瓜上分离到的烟草坏死病毒(TNV),其寄主范围较广,在藜科、苋科、豆科和茄科一些植物上呈局部坏死反应,并在葫芦科一些植物上呈局部坏死斑和系统花叶症状。病毒的热灭活点为90~95℃,稀释限点10-5~10-6,体外保毒期在10天以上。病毒颗粒呈六角形,直径约25毫微米。提纯病毒的紫外吸收,最高260毫微米,最低为245毫微米;沉降常数为94s。从病毒提取的核酸的紫外吸收,最高为260毫微米,最低234毫微米;经聚丙烯酰胺凝胶电泳测定,核酸分子量为1.4×10⁶;病毒蛋白的分子量为2.75×10⁴。经血清学试验,该病毒与烟草花叶病毒的A型和D型的抗血清发生沉淀反应,而与黄瓜坏死病毒的抗血清无沉淀反应。     

Watermelon necrosis caused by a strain of melon necrotic spot virus

A severe disease of watermelon (Citrullus vulgaris) grown in plastic houses in Crete, was characterized by leaf and stem necrosis followed by death of the plants. A strain of melon necrotic spot virus (MNSV) was identified as the causal agent of the disease on the basis of biological, morphological and serological properties. The watermelon strain of MNSV induced only local necrotic lesions in melon and cucumber plants and was serologically distinct from MNSV Cretan isolate. Gomphrena globosa was found a useful herbaceous host for differentiating it from common isolates of MNSV.     

甜菜潜隐病毒的研究

 从市场出售的甜菜(Beta vulgaris)种子中首次得到了直径约30毫微米的等轴病毒粒子的分离物。接种试验证明,它不能由摩擦接种传病,也不能由桃蚜(Myzus persicae)传播,但可由种子传毒,种子带毒率高达87.5%。受侵染的甜菜植株不表现症状,体内含毒量很低,病株汁液须经部分提纯并浓缩后才能在电镜下观察到病毒粒子和用琼脂双扩散法检测到病毒。在病株叶片的超薄切片中,未发现病毒粒子和细胞学的变异。在氯化铯溶液中,病毒粒子的浮力密度为1.37克/毫升左右。该分离物能与甜菜潜隐病毒抗血清产生沉淀反应,但与黄瓜花叶病毒无血清学关系。根据上述基本性状,该分离物归属于甜菜潜隐病毒(Beet Cryptic Virus)。     

京郊大蒜病毒病的研究及其鳞茎中病毒的脱除

 京郊大蒜病毒病发生普遍,主要症状为条纹花叶、矮化和叶片扭曲畸形。病体细胞中含大量线状病毒粒体和风轮状内含物。线状病毒粒体长度范围为250-1875nm,以长度550-800nm粒体居多。其中长700-800nm的粒体被鉴定为大蒜花叶病毒(GMV),回接脱毒大蒜叶片产生条纹花叶症状。血清学鉴定表明GMV与洋葱黄矮病毒(OYDV)有近缘关系。长度500-600nm的粒体可能为大蒜潜隐病毒(GLV),回接脱毒大蒜不产生花叶症状。下述两种病毒为京郊大蒜的主要病毒。此外,个别标样含烟草花叶病毒,但不是大蒜的主要病毒。对800nm以下线状病毒粒体归属尚待研究。
应用营养茎尖、生殖茎尖和根尖分生组织培养技术,可以脱去上述主要病毒,获得脱毒大蒜。     

河北省赤豆花叶病毒分离物的鉴定

 从河北省赤豆实生苗上获得一个分离物,它系种传的,引起赤豆产生疱状花叶,易经汁液摩擦接种,桃蚜(Myzus persicae)、豆蚜(Aphis cracivara)以非持久性方式传播,其钝化温度为55-60℃(十分钟)稀释限点为10^-2-10^-3,体外存活期为1-2天。寄主范围狭窄,系统侵染大豆(Glycine max)、绿豆(Phaselous aureus)、豇豆(Vigna sinensis cv.花豇豆)、棉豆(Phaseolus lunatus)、赤豆(P.angularis)等豆科植物,局部侵染苋色藜(Chenopodium amaranticolor)、昆诺藜(C.quinoa)和番杏(Tetragonia expensa)。病毒粒体线条形,略弯曲,755×13nm.A₂₆₀/A₂₈₀比值为1.224。病组织超薄切片中有风轮状内含体。SDS-免疫双扩散试验表明它与黑眼豇豆花叶病毒(BICMV)血清学关系十分密切。综合以上特征,我们认为赤豆花叶病毒河北省分离物为BICMV。河北省赤豆种子携带该病毒百分率为3-5%,它是田间主要毒源之一。     

A South African isolate of ryegrasss mosaic virus

An isolate of ryegrass mosaic virus (RMV-SA) was found in Italian ryegrass plants in two areas in South Africa. Preliminary characterization indicated that the virus exhibited the same symptoms and host range as RMV strains previously found in Canada and the UK. The virus was transmitted by the eriophyid mite Abacarus hystrix. Particle structure and size were comparable to the USA and UK strains. The coat protein and RNA were found to be slightly larger than previously reported, being 32kDa and 2·8 × 10⁶, respectively. The virus exhibited serological cross-reactivity with antisera to RMV-Wales and RMV-Canada but was not serologically related to other Rymovirus members.     

水蜡A病毒在呼和浩特市和哈尔滨市紫丁香上的发生及其基因组分子特征分析

为明确侵染紫丁香Syringa oblata并引起褪绿花叶症状的病毒种类及其基因组分子特征,利用透射电子显微镜对分离自呼和浩特市和哈尔滨市的紫丁香病样中的病毒粒子进行观察,并通过小RNA高通量测序和RT-PCR技术对其进行检测分析。结果表明,在紫丁香显症叶片的病毒粗提液中观察到长约600 nm、宽约13 nm的线状病毒粒子。利用小RNA高通量测序和RT-PCR技术从病样中检测到水蜡A病毒（Ligustrum virus A,LVA）,发病率为3.7%。呼和浩特市紫丁香分离物LVA-Sob的基因组序列全长8 525 nt,包含6个开放阅读框,分别编码Rep（1 968 aa）、TGB1（229 aa）、TGB2（107 aa）、TGB3（60 aa）、CP（294 aa）和NABP（119 aa）共6个蛋白。序列一致性分析表明,分离物LVA-Sob与韩国水蜡树分离物LVA-SK的基因组序列一致率高达97.9%,而与我国辽宁省暴马丁香分离物LVA-DX的基因组序列一致率仅为73.6%。在这3个LVA分离物基因组中没有检测到重组事件;基于基因组和cp基因序列的系统发育树显示这3个LVA分离物形成一个分支,并与瑞香S病毒（daphne virus S,DVS）有较近的亲缘关系。