Analysis of genetic relations between <Emphasis Type="Italic">Broad bean wilt virus 1</Emphasis> and <Emphasis Type="Italic">Broad bean wilt virus 2</Emphasis>

We determined the complete nucleotide sequence of RNA-1 and the 5-terminal region of RNA-2 from Broad bean wilt virus 1 (BBWV-1) isolate PV132. This report is the first analysis of the genome organization of BBWV-1. We also determined the complete nucleotide sequence of RNA-1 from Broad bean wilt virus 2 (BBWV-2) isolate IP and analyzed the genetic relations between BBWV-1 and BBWV-2. Similar to the BBWV-2 isolates, both RNAs of PV132 encoded a single large polyprotein, which was predicted to contain some functional proteins in a manner similar to those of comovirus. With respect to the deduced amino acid sequences of the mature proteins, PV132 and IP had only 20%–40% homology to comovirus. On the other hand, IP was 73%–98% homologous to BBWV-2 isolates, but PV132 was 39%–67% homologous to the isolates. Although the extent of the homologies differed, the homologies were limited between BBWV-1 and BBWV-2 not only for the coat protein but also for the other proteins. These results clearly support the placement of BBWV-1 and BBWV-2 in the genus Fabavirus as distinct species, proposed on the basis of double immunodiffusion tests.The nucleotide sequence data reported are available in the DDBJ/EMBL/GenBank databases under the accession numbers AB084450 (RNA-1 of isolate PV132), AB084451 (RNA-2 of isolate PV132), and AB023484 (RNA-1 of isolate IP)     

Incidence of viruses in <Emphasis Type="Italic">Alstroemeria</Emphasis> plants cultivated in Japan and characterization of <Emphasis Type="Italic">Broad bean wilt virus-2, Cucumber mosaic virus</Emphasis> and <Emphasis Type="Italic">Youcai mosaic virus</Emphasis>

Alstroemeria plants were surveyed for viruses in Japan from 2002 to 2004. Seventy-two Alstroemeria plants were collected from Aichi, Nagano, and Hokkaido prefectures and 54.2% were infected with some species of virus. The predominant virus was Alstroemeria mosaic virus, followed by Tomato spotted wilt virus, Youcai mosaic virus (YoMV), Cucumber mosaic virus (CMV), Alstroemeria virus X and Broad bean wilt virus-2 (BBWV-2). On the basis of nucleotide sequence of the coat protein genes, all four CMV isolates belong to subgroup IA. CMV isolates induced mosaic and/or necrosis on Alstroemeria. YoMV and BBWV-2 were newly identified by traits such as host range, particle morphology, and nucleotide sequence as viruses infecting Alstroemeria. A BBWV-2 isolate also induced mosaic symptoms on Alstroemeria seedlings.     

Broad bean wilt virus Causes Necrotic Symptoms and Generates Defective RNAs in Capsicum annuum

ABSTRACT A virus was isolated from hot pepper (Capsicum annuum cv. Hyang Chon) growing in Korea and displaying necrotic spots or streaks on leaves and stems followed by stunting and death of plants. Morphological and host range analyses of extracts from infected plants suggested that the causal agent of disease was a Broad bean wilt virus (BBWV), and the virus was tentatively named a Korean isolate of BBWV (BBWV-K). When the isolate was back-inoculated onto hot pepper plants, it induced symptoms similar to those of naturally infected hot pepper in the field. Two coat proteins (CPs) of 44 and 22 kDa, corresponding to a large CP and a small CP, respectively, were identified from the virus, and both reacted specifically with polyclonal antibody to BBWV 2. The complete nucleotide sequences of RNA 1 and RNA 2 of the isolate were determined from cDNA clones. The deduced amino acid sequence data from the putative proteins encoded by RNA 1 and 2 of the BBWV-K indicated a closer relationship with the isolates of BBWV 2 than BBWV 1. However, sequence comparison of the 5' noncoding regions of the viruses differentiates BBWV-K from other BBWV 2 isolates. Another distinctive feature of the BBWV-K is that it generates defective RNAs in hot pepper exhibiting necrotic symptoms, which is the first report of defective RNAs in the Fabavirus genera of BBWVs.     

Broad bean wilt virus 2 in yams

We identified Broad bean wilt virus 2 (BBWV-2) in yams based on particle morphology, test plant symptoms, protein features, aphid transmission, and molecular classification using nucleotide sequences of coat protein genes.     

侵染勿忘我花卉的蚕豆萎蔫病毒的鉴定

从勿忘我（Ｍｙｏｓｏｔｉｓｓｉｌｖａｔｉｃａ）上采集到１病株，接种昆诺阿藜，接种叶褪绿斑，系统顶枯，最后整株枯死。蚕豆接种叶呈黑色坏死斑，主茎变褐、全株萎蔫。矮牵牛接种叶褪绿斑，系统花叶。用诱捕修饰法制片，观察到病毒粒体球形，并在病毒粒体外面包被着一层抗体。琼脂双扩散试验呈阳性反应     

蚕豆萎蔫病毒RNA-2编码的多聚蛋白切割位点分析

 Broad bean wilt virus (BBWV) is the type species of the genus Fabavirus.     

Variability in Geminivirus Isolates Associated with Phaseolus spp. in Brazil

ABSTRACT Bean golden mosaic geminivirus (BGMV) is the single most devastating virus of common beans in the tropical and subtropical Americas and the Caribbean Basin. The BGMV from Brazil, named BGMV-BZ, is considered distinct from BGMV-PR isolates from Puerto Rico, Guatemala, and the Dominican Republic because of DNA sequence data, the ability to form pseudorecombinants, and mechanical transmissibility properties. In bean-growing areas of Brazil, samples were collected from beans, lima beans, and the weed Leonurus sibiricus displaying typical symptoms of infection by geminiviruses. Viral DNA fragments comprising part of the rep gene, the common region, and part of the cp gene were amplified by polymerase chain reaction, cloned, and sequenced. The bean samples had geminivirus with sequences nearly identical to that of BGMV-BZ collected in Goiania, state of Goiás, in 1986. The sample from lima bean contained a new species of geminivirus that induces symptoms similar to those induced by BGMV-BZ and was named lima bean golden mosaic virus (LBGMV-BR). While all sequences from bean samples clustered with BGMV-BZ, the sequence from the lima bean isolate stood alone. A mixed infection with abutilon mosaic geminivirus was also found in a single sample from the state of S?o Paulo. DNA sequence comparisons indicate that the virus isolate from L. sibiricus represents a new geminivirus species, designated here as leonurus mosaic virus.     

从葎草中检出复合侵染的多种病毒

采用抗原直接包被酶联免疫吸附测定法（ELISA）对采自重庆近郊的34个葎草样品进行了主要病毒种类的检测。其中马铃薯Y病毒(Potato virus Y, PVY)的侵染最普遍,其阳性检出率达44.12%;马铃薯X病毒(Potato virus X, PVX)的阳性检出率最低,仅为26.47%,其余5种病毒,烟草花叶病毒(Tobacco mosaic virus, TMV)、黄瓜花叶病毒(Cucumber mosaic virus, CMV)、番茄花叶病毒(Tomato mosaic virus, ToMV)、芜菁花叶病毒(Turnip mosaic virus, TuMV)及蚕豆萎蔫病毒2号(Broad bean wilt virus 2, BBWV-2)的阳性检出率均为35.29%。葎草样品受多种病毒的复合侵染现象非常严重,15个阳性样品中病毒复合侵染率为80%,其中75%的样品检测到7种病毒复合侵染。     

我国太子参主产区病毒病发生情况调查

为调查我国太子参主产区病毒病发生情况,使用RT-PCR法对采自福建省柘荣县,贵州省施秉县、丹寨县和安徽省宣城市的71份具有典型病毒病症状的太子参样品进行检测,并根据CP氨基酸序列对不同地区病毒进行系统进化分析。RT-PCR结果显示,65份样品检出病毒,检出率为91.55%,其中,56份样品检出芜菁花叶病毒Turnip mosaic virus(TuMV),49份样品检出蚕豆萎蔫病毒2号Broad bean wilt virus 2(BBWV2),9份样品检测出黄瓜花叶病毒Cucumber mosaic virus(CMV),检出率分别为78.87%、69.01%和12.68%。未检测到烟草花叶病毒Tobacco mosaic virus(TMV)。序列和系统发育进化分析显示,本研究获得的TuMV、BBWV2和CMV与NCBI数据库中的序列相似度较高,核苷酸相似度分别为98.05%～98.98%、93.61%～94.25%和98.02%～99.17%,氨基酸序列相似度分别为:96.14%～97.47%、97.33%～98.50%和98.00%～100%,分别属于World-B组、Ⅰ-a亚...     

豌豆病毒病病原研究

 1986年至1990年,从豌豆田中采集了150余份病毒病样本,鉴定出蚕豆萎蔫病毒(BB-WV)、芜菁花叶病毒(TuMV)、马铃薯Y病毒组分离物、黄瓜花叶病毒(CMV)、莴苣花叶病毒(LMV)、大豆花叶病毒(SMV)、豌豆花叶病毒(PMV)、菜豆黄花叶病毒(BYMV)和苜蓿花叶病毒(AMV)等9种病毒。样本中,BBWV所占的比例最高,达59.2%,其次为CMV,占15.5%。BBWV常与CMV复合侵染豌豆,LMV发生也较普遍。田间调查表明,豌豆病毒病发病率因种植地区及品种不同而有差异,平均发病率为12.4%。     

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。     

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.     

蚕豆萎蔫病毒2号分离物PV131和P158侵染寄主植物的细胞病理比较观察

 透射电镜观察比较了蚕豆萎蔫病毒2号(BBWV-2)分离物PV131和P158侵染昆诺藜(Chenopodium quinoa)和蚕豆(Vicia faba)的细胞超微结构变化。结果显示:受PV131分离物侵染的昆诺藜叶肉细胞质中膜结构大量增生,形成有膜状结构、大小泡囊及电子致密物质的特殊膜增生区;病毒粒子散布在细胞质中,形成大块结晶体和管状结构,该管状结构直径约75nm,横切面由9个病毒粒子组成;受PV131侵染的蚕豆叶肉细胞中也形成膜增生区和管状结构,未见病毒结晶体。受P158分离物侵染的昆诺藜和蚕豆叶肉细胞病变情况与PV131分离物相似,形成膜增生区和相同的管状结构,在蚕豆叶肉细胞中也未观察到病毒结晶体。上述结果表明:2个BBWV-2分离物虽然在不同寄主植物上引起的细胞病变程度有差异,但其细胞病理学特征是由病毒基因结构所决定,与寄主的种类无关。     

青蒿中番茄斑萎病毒和烟草花叶病毒的分子鉴定及相关序列分析

番茄斑萎病毒(tomato spotted wilt virus, TSWV)和烟草花叶病毒(tobacco mosaic virus, TMV)是2种重要的植物病原病毒, 对多种经济作物的产量和品质均造成严重影响。2021年-2022年, 在云南省丽江市烟草种植区不同烟区采集叶片黄化、皱缩以及无症状的青蒿Artemisia caruifolia样品共计14份, 利用免疫金标速测卡和RT-PCR对其病原病毒进行检测。利用免疫金标速测卡检测结果显示, 在所检样品中有9份样品检测出TSWV, 检出率为64.28%, 有3份样品检测出TMV, 检出率为21.43%, 2种病毒复合侵染的检出率同样为21.43%;利用RT-PCR对复合侵染的3份样品进行分子检测, 结果显示, 在3份复合侵染青蒿样品中获得3条TSWV N基因序列、3条TMV cp基因序列和2条TMV RdRp部分序列。TSWV青蒿分离物与分离自云南的TSWV-2分离物相似性最高, 为99.6%;TMV青蒿分离物与分离自辽宁的TMV-Shenyang分离物和分离自云南的TMV-Yongren-1相似性最高, 均大于99.4%。这是首次发现TSWV和TMV 2种不同属病毒复合侵染青蒿。     

Molecular and biological characterization of Macroptilium yellow mosaic virus from Jamaica

The molecular and biological characterization of a begomovirus infecting the common weed Macroptilium lathyroides from Jamaica are reported. The virus showed 92% sequence identity to an isolate of Macroptilium yellow mosaic virus (MaYMV) from Cuba, but was distinct from the two other begomoviruses isolated from M. lathyroides , namely Macroptilium yellow mosaic Florida virus (80% identity) and Macroptilium mosaic Puerto Rico virus (68% identity). Hence, the Jamaican begomovirus was considered an isolate of MaYMV and called Macroptilium yellow mosaic virus -[Jamaica] (MaYMV-[JM]). In infectivity studies using cloned DNA-A and DNA-B genomic components, MaYMV-[JM] infected red kidney bean ( Phaseolus vulgaris ) and produced mild symptoms in Scotch Bonnet pepper ( Capsicum chinense ), but did not infect cabbage ( Brassica oleracea ). This information has implications for the development of strategies to control begomovirus diseases in Jamaica and elsewhere.     

Two Newly Described Begomoviruses of Macroptilium lathyroides and Common Bean

ABSTRACT Macroptilium lathyroides, a perennial weed in the Caribbean region and Central America, is a host of Macroptilium yellow mosaic Florida virus (MaYMFV) and Macroptilium mosaic Puerto Rico virus (MaMPRV). The genomes of MaYMFV and MaMPRV were cloned from M. lathyroides and/or field-infected bean and the DNA sequences were determined. Cloned A and B components for both viruses were infectious when inoculated to M. lathyroides and common bean. Comparison of the DNA sequences for cloned A and B components with well-studied begomovirus indicated that MaMPRV (bean and M. lathyroides) and MaYMFV (M. lathyroides) are unique, previously undescribed begomo-viruses from the Western Hemisphere. Phylogenetic analysis of viral A components indicated that the closest relative of MaYMFV are members of the Bean golden yellow mosaic virus (BGYMV) group, at 76 to 78% nucleotide identity, whereas the closest relative for the A component of MaMPRV was Rhynchosia golden mosaic virus at 78% nucleotide identity. In contrast, BGYMV is the closest relative for the B component of both MaYMFV and MaMPRV, with which they share approximately 68.0 and approximately 72% identity, respectively. The incongruent taxonomic placement for the bipartite components for MaMPRV indicates that they did not evolve entirely along a common path. MaYMFV and MaMPRV caused distinctive symptoms in bean and M. lathyroides and were transmissible by the whitefly vector and by grafting; however, only MaYMFV was mechanically transmissible. The experimental host range for the two viruses was similar and included species within the families Fabaceae and Malvaceae, but only MaYMFV infected Malva parviflora and soybean. These results collectively indicate that MaMPRV and MaYMFV are new, previously undescribed species of the BGYMV group, a clade previously known to contain only strains and isolates of BGYMV from the Caribbean region that infect Phaseolus spp. Both MaYMFV and MaMPRV may pose an economic threat to bean production in the region.     

引起北京豇豆系统坏死和花叶的一个分离物——Ⅰ.寄主范围、提纯和超薄切片

 1981年夏,在京郊大面积受害的豇豆上分离到一株毒原一豇豆分离物。其症状是顶叶系统花叶,褐色坏死和畸形。生物学测定证明其寄主范围广泛,在所测试的6个科19种植物中,能侵染5个科的十余种植物。苋色藜、奎宁藜、千日红、曼陀罗、蚕豆、豇豆为其易感寄主。但不能侵染白菜、油菜、芜菁、黄瓜、菜豆、豌豆、普通烟和心叶烟等植物。该分离物虽在蚕豆上病状与蚕豆萎蔫病毒Stubbs典型株相似,但从其不侵染普通烟和心叶烟似乎又不完全相同。稀释限点:10^-3~10^-4;热灭活点50~55℃;体外保毒期(室温)4天以上。桃蚜或豆蚜传。提纯病毒颗粒呈直径25mm,六角形,其三种组分之沉降常数分别为50S (T),67S (M)和118S (B)。提纯的病毒制品其颗粒极易聚集特别是盐类存在时。电镜观察奎宁藜局部退绿斑的超薄切片发现为蚕豆萎蔫病毒所具有的特征性的横截面由7~9个病毒颗粒所组成的管状结晶物以及由非病毒颗粒所组成的大型晶体内含体。根据上述寄主范围、病状、病毒颗粒形态和组分以及内含体的特征,豇豆分离物是蚕豆萎蔫病毒,很可能是与已知的四种毒株不同的豇豆株。     

Biological, serological, and molecular variabilities of clover yellow vein virus

ABSTRACT A comparative study was made on the host reactions, serological properties, and nucleotide sequences of the coat protein (CP) gene of 10 clover yellow vein virus (C1YVV) isolates and one bean yellow mosaic virus (BYMV) isolate collected from different host plant species and locations in Japan. Two strains of C1YVV isolates, grouped on the basis of host reactions on Chenopodium amaranticolor, C. quinoa, Nicotianaclevelandii, N. benthamiana, Vicia faba, and Trifolium repens, corresponded to two serotypes determined by double-antibody sandwich- and triple-antibody sandwich-enzyme-linked immunosorbent assay using three polyclonal and nine monoclonal antibodies. These results were also confirmed by nucleotide sequence analysis of the CP gene. The CP gene of C1YVV isolates of strain 1, including the Australian isolate C1YVV-B, had 93 to 98% nucleotide identities and 97 to 99.6% amino acid identities. The CP of C1YVV isolates of strain 2, including the New Zealand isolate C1YVV-NZ, had 92 to 98% nucleotide identities and 95 to 98% amino acid identities. The nucleotide identities and the amino acid identities between the two C1YVV strains were 82 to 84%, and 90 to 94%, respectively. When compared with the CP sequences of 12 C1YVV isolates, the CP sequence of the BYMV isolate had 71 to 73% nucleotide identity and 73 to 77% amino acid identity. Amino acid sequence differences among C1YVV isolates from strains 1 and 2 were located mostly at the N-terminal regions of the CP. Our results indicated that the C1YVV isolates studied could be separated into two strains on the basis of host reactions, serology, and the nucleotide sequence of the CP gene.     

First report of <Emphasis Type="Italic">Bean common mosaic virus</Emphasis> in yam bean [<Emphasis Type="Italic">Pachyrhizus erosus</Emphasis> (L.) Urban] in Indonesia

Severe mosaic with leaf malformation and green vein banding was observed on yam bean in West and Central Java, Indonesia. Virions of the causal virus were flexuous filaments, about 700 nm in length, with a coat protein of 30 kDa. The virus was transmitted by mechanical inoculation and by aphids in a nonpersistent manner. The nucleotide sequence of the coat protein gene had the highest identity with that of Bean common mosaic virus (BCMV, genus Potyvirus) isolate VN/BB2-5. Based on demarcation criteria, including the genome sequence and host range, we tentatively designate this isolate as BCMV-IYbn (Indonesian yam bean). The nucleotide sequence reported is available in the DDBJ/EMBL/GenBank databases under accession number AB289438.