Pfaffia mosaic virus: a new potyvirus found infecting Pfaffia glomerata in Brazil

In Brazil plants of Pfaffia glomerata with mosaic symptoms were found to be infected with a previously undescribed potyvirus, Pfaffia mosaic virus (PfMV). Virus particles were long and flexuous, c.  10 × 700–800 nm, and cylindrical inclusions typical of potyviruses were present in cells of infected tissue. Partial host-range studies revealed that in addition to P. glomerata , PfMV infected only Chenopodium amaranticolor and Chenopodium quinoa . It was efficiently transmitted by the aphids Aphis gossypii and Myzus persicae . Polyclonal antiserum produced against the PfMV coat protein (CP) reacted with Potato virus Y (PVY), but not with four other potyviruses in PTA-ELISA. The similarity of the nucleotide sequence of the PfMV coat-protein gene ( CP ) varied from 7 to 76% when compared with other members of the family Potyviridae . Similarity of the 3' NTR sequence varied from 4 to 23%. In both cases the highest similarity was with PVY. These data indicate that PfMV is a new species in the genus Potyvirus .     

Characterization of potato Y potyvirus isolates from tomato crops in Islas Canarias (Spain)*

A disease caused by potato Y potyvirus (PVY) affects tomato plantations with variable severity in Tenerife Island. Affected plants show diverse symptoms such as necrotic lesions or mild to severe mosaic in leaves and whitish spots in green fruits that remain after ripening. Tomato PVY isolates and few potato and capsicum PVY isolates have been characterized on the basis of biological, serological and molecular criteria. All PVY isolates reacted positively to monoclonal antibodies specific for PVY^O/C or PVY^N strains, and nearly 50% of tomato PVY isolates were recognized by both. Differentiation of PVY strains according to the response of inoculated experimental plants was confusing due to the variability of viral aggressiveness and symptomatology induced. RFLP analysis of the CP gene and 3’untranslated region (UTR) revealed high variability. In addition to mixed infection by different PVY strains, the biological and molecular properties of those tomato PVY isolates that react to both monoclonal antibodies could be explained as the result of RNA recombination between distinct PVY strains which infect the same host plant.     

Biological, serological, and molecular differences among isolates of potato a potyvirus

ABSTRACT Sequences of the coat protein (CP) and 3'-end nontranslated region (3'NTR) of 13 isolates and the helper component proteinase (HC) of nine isolates of potato A potyvirus (PVA) were determined and compared with the eight previously determined PVA CP and 3'NTR sequences and one HC sequence. CP amino acid (aa), 3'NTR nucleotide, and HC aa sequence identities were 92.9, 93.4, and 94.8%, respectively. Sequence data, serological tests, and the necrotic local lesions induced in the leaves of the potato hybrid 'A6' confirmed that tamarillo mosaic virus is a strain of PVA. The aa substitutions A6T and G7S in the CP N-terminus were correlated with loss of aphid transmissibility. Development of necrotic lesions or nonnecrotic symptoms in the systemically infected leaves or lack of systemic spread in potato cv. King Edward were used to place the PVA isolates into four strain groups, but this grouping was not correlated with any differences in CP, HC, or 3'NTR. Recognition of CP by three monoclonal antibodies was used to place the PVA isolates into three groups different from the four groups above. The epitopes of two mono-clonal antibodies were mapped by site-directed mutagenesis to the same lysine residue at the CP aa 34.     

我国12省市玉米矮花叶病病原鉴定及病毒致病性测定

 利用甘蔗花叶病毒(Sugarcane mosaic virus,SCMV)单克隆抗体细胞株2B5腹水和SCMV、玉米矮花叶病毒(Maize dwarf mosaic virus,MDMV)、高粱花叶病毒(Sorghum mosaic virus,SrMV)和约翰逊草花叶病毒(Johnsongrass mosaic virus,JGMV)的特异性引物对我国浙江、江苏、上海、山东、河南、河北、北京、山西、陕西、甘肃、四川、云南12省市15个地点的176株玉米矮花叶病病样分别进行了间接ELISA和免疫捕获反转录PCR (IC-RT-PCR)检测,结果表明这些病样均含有SCMV,而无MDMV、SrMV或JGMV存在,表明上述12省市的玉米矮花叶病病原为SCMV。进一步对甘肃(GS)、四川(SC)、云南(YN)3个SCMV分离物的近全长CP基因进行了序列测定,并测定了浙江分离物(ZJ)和甘肃分离物(GS)在13个玉米品种上的致病性。     

Viruses of vegetable crops in Albania

Field surveys were carried out in the main vegetable-growing areas of Western and Central Albania to evaluate the sanitary status of open-field and protected cultivations of capsicum, tomato, potato, watermelon, cucumber, courgette, aubergine, lettuce, cabbage, chicory, leek and celery. The following viruses were detected: Alfalfa mosaic alfamovirus (AMV), Cucumber mosaic cucumovirus (CMV), Potato Y potyvirus (PVY), Tomato spotted wilt tospovirus (TSWV) and Watermelon mosaic potyvirus 2 (WMV-2). The virus found most frequently was CMV and all the isolates identified were of subgroup IA. AMV was also detected in several areas and all isolates were of subgroup II, suggesting a French origin. Finally, at the time of this survey, TSWV infections appeared to be moderately relevant and absent in protected crops. This is the first documented record of AMV, CMV, PVY, TSWV and WMV-2 in Albania. No infection by Tomato yellow leaf curl begomovirus (TYLCV) or Zucchini yellow mosaic potyvirus (ZYMV) was detected during this survey.     

Genetic diversity of Potato virus Y infecting tobacco crops in China

Genetic variability of Potato virus Y (PVY) isolates infecting potato has been characterized but little is known about genetic diversity of PVY isolates infecting tobacco crops. In this study, PVY isolates were collected from major tobacco-growing areas in China and single-lesion isolates were produced by serial inoculation on Chenopodium amaranticolor. Most isolates (88%) caused systemic veinal necrosis symptoms in tobacco. Of these, 16 isolates contained a PVY(O)-like coat protein (CP) and PVY(N)-like helper component proteinase (HC-pro) and, in this respect, were similar to the PVY(N-Wi), PVY(N:O), and PVY-HN2 isolates characterized from potato in Europe, the United States, and China, respectively; two isolates contained a PVY(O)-like HC-pro and a PVY(N)-like CP; another two isolates had recombination junctions in the CP-encoding region. Both the HC-pro and CP of PVY were under negative selection as a whole; however, seven amino acids in HC-pro and six amino acids in CP were under positive selection. Selection pressures differed between the subpopulations of PVY distinguished by phylogenetic analysis of HC-pro and CP sequences. When PVY isolates from potato were included, no host-specific clustering of the PVY isolates was observed in phylogenetic and nucleotide diversity analyses, suggesting frequent spread of PVY isolates between potato and tobacco crops in the field.     

Serological Analysis and Coat Protein Sequence Determination of Potato Virus Y (PVY) Pepper Pathotypes and Differentiation from Other PVY Strains

The serological relationships of Potato Virus Y (PVY) isolates belonging to the pepper pathotypes 0, 1 and 1-2 were established by enzyme-linked immunosorbent assay (ELISA). PVY pepper pathotypes did not react with monoclonal antibodies which typically recognize non-pepper strains within the PVY group, leading to discrimination between these two groups of strains. No serological differences were found between the three PVY pepper pathotypes. The coat protein (CP) nucleotide and predicted amino acid sequences of the three different PVY pepper pathotypes were determined. The highest sequence similarity was found between pathotypes 0 and 1 (99.2%), while the lowest occurred between these two and pathotype 1–2 (98.1%). PVY strains from potato and tobacco appeared more distantly related. Phenetic analysis of the CP amino acid sequences showed that the PVY pepper pathotypes formed a tightly clustered group separate from other PVY strains.     

Identification and characterization of a potyvirus causing chlorotic spots on <Emphasis Type="Italic">Phalaenopsis</Emphasis> orchids

A putative virus-induced disease showing chlorotic spots on leaves of Phalaenopsis orchids was observed in central Taiwan. A virus culture, phalaenopsis isolate 7-2, was isolated from a diseased Phalaenopsis orchid and established in Chenopodium quinoa and Nicotiana benthamiana. The virus reacted with the monoclonal antibody (POTY) against the potyvirus group. Potyvirus-like long flexuous filament particles around 12–15 × 750–800 nm were observed in the crude sap and purified virus preparations, and pinwheel inclusion bodies were observed in the infected cells. The conserved region of the viral RNA was amplified using the degenerate primers for the potyviruses and sequence analysis of the virus isolate 7-2 showed 56.6–63.1% nucleotide and 44.8–65.1% amino acid identities with those of Bean yellow mosaic virus (BYMV), Beet mosaic virus (BtMV), Turnip mosaic virus (TuMV) and Bean common mosaic virus (BCMV). The coat protein (CP) gene of isolate 7-2 was amplified, sequenced and found to have 280 amino acids. A homology search in GenBank indicated that the virus is a potyvirus but no highly homologous sequence was found. The virus was designated as Phalaenopsis chlorotic spot virus (PhCSV) in early 2006. Subsequently, a potyvirus, named Basella rugose mosaic virus isolated from malabar spinach was reported in December 2006. It was found to share 96.8% amino acid identity with the CP of PhCSV. Back-inoculation with the isolated virus was conducted to confirm that PhCSV is the causal agent of chlorotic spot disease of Phalaenopsis orchids in Taiwan. This is the first report of a potyvirus causing a disease on Phalaenopsis orchids.     

Further Evidence that Shallot Yellow Stripe Virus (SYSV) Is a Distinct Potyvirus and Reidentification of Welsh Onion Yellow Stripe Virus as a SYSV Strain

ABSTRACT An antiserum to shallot yellow stripe virus (SYSV) was raised and used in combination with a range of other antisera to potyviruses of Allium spp. in electron microscopic decoration experiments. The serological results corroborated an earlier finding that the type isolates of SYSV and Welsh onion yellow stripe virus (WoYSV) are closely related to each other and only distantly related to onion yellow dwarf (OYDV) and leek yellow stripe (LYSV) viruses, the two other major potyviruses infecting Allium spp. Moreover, the decoration results indicated that Japanese potyviruses named OYDV and Wakegi yellow dwarf virus are isolates of SYSV. Sequence analysis of the 3'-terminal regions of the SYSV and WoYSV ge-nomes revealed coat protein (CP) amino acid and 3'-nontranslated region (3'-NTR) nucleotide sequence identities of 95 and 89%, respectively. The CP amino acid and 3'-NTR nucleotide sequences of these viruses differed from those of OYDV and LYSV by >25 and >67%, respectively. The serological and molecular studies showed that SYSV and WoYSV are different strains of a potyvirus distinct from OYDV and LYSV. For priority reasons, we propose that these strains together with the Wakegi-type isolates of OYDV described in Japan be referred to as SYSV and that SYSV isolates from Allium spp. other than shallot be designated as the Welsh onion strain of SYSV (SYSV-Wo).     

Sequence of the 3′-terminal region of the RNA of a mite transmitted potyvirus fromHordeum murinum L.

The 3026 nucleotides upstream of the 3-polyadenylated tract of a mite transmitted virus fromHordeum murinum L. were cloned and sequenced, and portions of the sequence were expressed inEscherichia coli. Sequence comparisons with wheat streak mosaic virus (WSMV), Agropyron mosaic virus (AgMV) and Hordeum mosaic cirus (HoMV), three mite transmitted potyviruses, and potato virus Y (PVY), the type member of the genusPotyvirus, revealed that the virus is probably a potyvirus, but distinct from WSMV, AgMV, HoMV, and PVY. Serological tests further demonstrated these differences and that the virus is serologically related to another potyvirus, brome streak mosaic virus (BrSMV). We conclude that the virus should be named as the Hordeum isolate of BrSMV.     

Identification of the principal viruses infecting tomato crops in Tunisia

Serological tests have been used to detect viruses associated with tomato in 257 samples collected in different regions in Tunisia, Cap-Bon, Sahel and South during successive seasons. The viruses detected were cucumber mosaic cucumovirus (CMV) and tomato aspermy cucumovirus (TAV), potato Y potyvirus (PVY), tobacco etch potyvirus (TEV) and pepper veinal mottle potyvirus (PVMV), tomato mosaic tobamovirus (ToMV) and tobacco mosaic tobamovirus (TMV), tobacco rattle tobravirus (TRV), alfalfa mosaic alfamovirus (AMV), tomato spotted wilt tospovirus (TSWV), tomato ringspot nepovirus (TomRSV) and potato X potexvirus (PVX). Some were detected in all three regions surveyed, at variable frequencies: TMV, CMV, TEV, PVY, ToMV, AMV, TAV, TSWV and TRV. Others were only detected in two regions (PVMV in Cap-Bon and Sahel and PVX in Sahel and in the south) or one region (TomRSV in Cap-Bon). Movement of individual viruses from one region to another may be due to movement of specific vectors, as in the case of the thrips-transmitted TSWV moving from the south to the north. Some of these viruses were found for the first time in Tunisia.     

小麦黄色花叶病毒不同分离物外壳蛋白(CP)基因序列的比较分析

 核苷酸序列分析结果表明,小麦黄色花叶病毒(W YMV)不同分离物的外壳蛋白基因存在一定的差异。邓州分离物CP基因在其31~33nt处均缺失了3个核苷酸,其余分离物与潢川分离物及日本分离物长度一致,均为882nt。不同分离物CP基因核苷酸序列同源性为97.3%~98.9%,由此推导的氨基酸序列同源性为97.6%~99.3%,外壳蛋白N末端的110个氨基酸和C末端的55个氨基酸在各个分离物间是高度保守的。潢川分离物有5个氨基酸与其它5个分离物明显不同。WYMV不同分离物外壳蛋白序列分析结果进一步确认了WYMV与WSSMV为Bymovirus属的2种不同病毒。     

Molecular Characterization of Potato virus V Genomes from Europe Indicates Limited Spatiotemporal Strain Differentiation

ABSTRACT Because there were no previous reports on the molecular characterization of Potato virus V (PVV, genus Potyvirus, family Potyviridae), the complete genomic sequence of PVV isolate Dv42 was determined. The length of the single-stranded messenger-polarity RNA genome was 9,851 nt (nucleotides), followed by a poly(A) tail. The genome contained a 5'-terminal nontranslated region (5'-NTR; 204 nt), a single open reading frame (nucleotides 205-9406; 3,067 amino acids), and a 3'-NTR that was unusually long (446 nt) compared with that of Potato virus Y (PVY; 331-nt 3'-NTR), Potato virus A (PVA; 207-nt 3'-NTR), and other potyviruses that naturally infect Solanaceae species. Phylogenetic analysis with the cylindrical inclusion protein-encoding and coat protein (CP)-encoding regions indicated that PVV Dv42 was most closely related to Pepper mottle virus and PVY, respectively. Seven PVV isolates (including Dv42) collected from cultivated potatoes in the Netherlands, the United Kingdom, and Norway from 1964 to 1997 were uniform in serological properties and symptomatology in indicator hosts that could distinguish strains of PVY and PVA. The nucleotide sequences of the 5'-NTR, P1, CP, and 3'-NTR regions of the PVV isolates were determined and were 94.6 to 99.5, 96.3 to 98.8, 96.4 to 98.7, and 96.3 to 99.6% identical, respectively. The amino acid similarities for the P1 and CP were 95.8 to 98.6 and 96.0 to 97.8%, respectively. Phylogenetic analysis of the CP sequences of PVV revealed no significant grouping, in contrast to PVY and PVA, which were grouped largely according to the previously recognized strains based on host responses. However, the relatively few differences in the P1 sequences of PVV were correlated with the different countries of origin. Hence, the PVV isolates infecting potatoes in Europe seem to vary little genetically and may belong to a single strain.     

Characterization of a strain of clover yellow vein potyvirus infecting borage (Borago officinalis L.) in Spain

A disease of borage ( Borago officinalis ) in Spain, characterized by severe mosaic and deformation of the leaves, was shown to be caused by a potyvirus. The borage-infecting potyvirus was characterized biologically by the symptoms induced in 23 indicator species and was shown to be transmitted experimentally by the aphid Myzus persicae in a non-persistent manner. In order to classify the borage-infecting potyvirus we have cloned and sequenced the entire coat protein gene and 3' non-coding region of the viral RNA. By comparing this nucleotide sequence with those of other members of the Potyviridae , we can identify the Spanish borage-infecting potyvirus as an isolate of clover yellow vein virus (CYVV), a virus so far only known to cause important diseases in forage legumes. This is the first record of CYVV in Spain and of CYVV infecting a natural host of the Boraginaceae.     

甜菜花叶病毒新疆分离物基因组3'末端序列分析

甜菜花叶病毒（Beet mosaic virus，BtMV）属马铃薯Y病毒科、马铃薯Y病毒属，可经多种蚜虫以非持久性方式传播，病毒粒子为弯曲线状，核酸为单分子正义ssRNA。目前只有美国华盛顿分离物的全序列以及斯洛伐克和英国少数几个分离物3’端的部分序列被报道。美国分离物全长9591 nt，3’端具有PolyA尾，编码一个由3086个氨基酸组成的多聚蛋白，与其它Potyvirus病毒一样可切割成10个蛋白，从N到C端依次为P1、HC—Pro、P3、6K1、CI、6K2、NIa—Vpg、NIa～Pro、NIb和CP。对于我国发生的BtMV，1981年Liu等报道了发生于北京地区菠菜上的BtMV，之后研究人员相继报道了黑龙江、内蒙古和新疆等甜菜主产区甜菜花叶病的发生及危害情况，并陆续开展了对BtMV的生物学特性、外壳蛋白分子量测定和氨基酸组分分析、细胞病理学等研究，目前对于我国发生的BtMV的分子结构特征还未见报道。本文报道了甜菜花叶病毒新疆分离物（BtMV—XJ）3’端的核酸序列，并与国外已报道序列进行了比较分析，为从分子水平上明确我国BtMV的分子结构特点、深入研究其编码蛋白的功能打下了基础。     

一株PVYNTN-NW黑龙江马铃薯分离物的检测鉴定

 马铃薯Y病毒(Potato virus Y,PVY)是马铃薯、烟草等茄科作物上的重要病毒,在与寄主共同进化过程中产生了许多株系。本文从黑龙江马铃薯样品中得到PVY分离物A12。ELISA结果表明A12被PVY^O的单克隆抗体特异性识别。A12开放阅读框为9 186个核苷酸,编码3 061个氨基酸,与SYR-II-Be1分离物的核苷酸和氨基酸序列一致率均最高,分别为98.3%和99.2%。系统发育分析发现A12与PVY^NTN-NW株系SYR-II型的分离物聚类到一起;重组分析表明,A12是N-605和O_z的重组体,重组类型与SYR-II-Be1相同。综合以上结果表明,A12属于PVY^NTN-NW株系SYR-II型。但与常见PVY^NTN-NW株系分离物在珊西烟引起叶脉坏死不同,A12产生花叶症状。A12辅助成分-蛋白酶在182位和245位的氨基酸均为精氨酸,而其它PVY^NTN-NW株系分离物为赖氨酸。本研究结果可为黑龙江马铃薯PVY的早期检测和有效防控提供理论指导。