Characterization of a potyvirus isolated from Tradescantia fluminensis in northern Italy

Summary. We isolated a potyvirus from Tradescantia fluminensis that was causing leaf distortion and mild mosaic. We cloned and sequenced a 1500 bp cDNA obtained by RT-PCR corresponding to the 3′ proximal region of the genome. We determined the host range and tested a series of potyviral antisera against our tradescantia virus isolate by immuno-enzymatic methods. Based on our results, we suggest that our viral isolate could be considered a new potyvirus species named Tradescantia mild mosaic potyvirus. Phylogenetic analysis confirmed that Tradescantia mild mosaic virus belongs to the genus Potyvirus within the family Potyviridae, but the virus could not be assigned to any of the potyvirus groupings recently defined.     

Konjak mosaic virus: the complete nucleotide sequence of the genomic RNA and its comparison with other potyviruses

Summary. Konjak mosaic virus (KoMV) belongs to the genus Potyvirus, family Potyviridae. The complete nucleotide sequence of KoMV F isolate (KoMV F) was determined. The genome is 9,544 nucleotides long excluding the 3′ terminal poly A tail and encodes a typical potyviral 350-kDa polyprotein of 3,087 amino acids. Phylogenetic analysis using known potyvirus polyproteins shows that KoMV constitutes a branch with yam mosaic virus, close to another branch including Japanese yam mosaic virus, turnip mosaic virus, scallion mosaic virus and lettuce mosaic virus. The 3′ terminal 1,842 nucleotides of a different isolate of KoMV, K-2, was also determined, covering the C-terminal 292 amino acids of the nuclear inclusion protein b (NIb), coat protein (CP), and the 3′ untranslated region. The amino acid sequences of the KoMV F CP and the nucleotide sequences of the KoMV F 3′ untranslated region showed 92.5 and 90.5% identity to the corresponding genes of K-2, 88.7–96.8 and 92.7–94.4% to those of Zantedeschia mosaic virus (ZaMV) isolates, 87.5–89.7% and 85.5–90.3% to those of Japanese hornwort mosaic virus (JHMV) isolates. These results showed that KoMV is a distinct potyvirus and that KoMV, ZaMV, and JHMV are members of the same potyvirus species. Considering that KoMV was the first of these to be described, ZaMV and JHMV may be considered isolates of KoMV.     

Daphne mosaic virus (DapMV), a new potyvirus from <Emphasis Type="Italic">Daphne mezereum</Emphasis> in the Czech Republic

Summary. Daphne shrubs with light green rings and mosaic on leaves contained flexuous filamentous virions (696 × 13 nm) and cylindrical inclusions typical of the subdivision III of Edwardson’s classification for inclusions induced by members of the family Potyviridae. Decoration tests using antisera to 67 potyviruses revealed distant serological relations among chilli veinal mottle virus, Colombian datura virus, papaya ringspot virus, tobacco vein mottling virus and yam mosaic virus. The 3′ terminal region of the virus genome was amplified by RT-PCR using primers specific for cloned and sequenced members of the family Potyviridae. The most similar sequences in the GenBank were those of isolates of wild potato mosaic virus (WPMV) and yam mild mosaic virus (YMMV), originating from Peru and Guadeloupe, respectively. The new sequence had 63.2% and 61.9% nucleotide identity to WPMV and YMMV in the coat protein gene. The results suggest that the Czech isolate from daphne should be regarded as a new member of the genus Potyvirus. The name daphne mosaic virus (DapMV) is suggested for this virus.     

The complete genome sequence of <Emphasis Type="Italic">Triticum</Emphasis> mosaic virus,a new wheat-infecting virus of the High Plains

The genome of Triticum mosaic virus (TriMV), a recently discovered mite-transmitted wheat potyvirus, was sequenced, characterized, and compared to other members of the family Potyviridae. TriMV has a single mRNA strand of 10,266 nucleotides with a predicted polyprotein consisting of 3,112 peptides. Protein alignments of the coat protein demonstrate that TriMV has 45.9% identity to Sugarcane streak mosaic virus strain AP (SCSMV-AP), but shares only 23.2% identity to Wheat streak mosaic virus. Although TriMV is mite-transmitted and could be placed in the genus Tritimovirus, it is significantly divergent and should be placed in the newly proposed genus Susmovirus.     

The 3′terminal sequence of Chinese yam necrotic mosaic

Summary.  The nucleotide sequence of the 3′terminal 1,905 residues of the Chinese yam necrotic mosaic virus (ChYNMV) RNA genome was determined. It contains one long open reading frame, which consists of 1,671 nucleotides encoding a protein of 557 amino acid residues. A partial amino acid sequence of the coat protein determined from purified ChYNMV particles was identical to the portion of the amino acid sequence deduced from the determined nucleotide sequence, which suggests that the nucleotide sequence includes the coat protein gene. Surprisingly, a homology search using the deduced amino acids sequence of the coat protein revealed that ChYNMV is closely related to the genus Maclura-virus within the family Potyviridae, although the virus has long been considered to be a carlavirus. Identification of cylindrical cytoplasmic inclusions, which are characteristic of the family Potyviridae, in ChYNMV-infected Chinese yam cells, as well as the morphology and length (660 nm) of the purified virus particles, support including the virus as a tentative new member of the genus Macluravirus. Received September 1, 2000/Accepted March 17, 2001     

A new potyvirus first isolated and identified from <Emphasis Type="Italic">Angelica sinensis</Emphasis>

A filamentous virus was isolated in Angelica sinensis (Angelica sinensis (Oliv.) Diels) which shows mosaic symptoms on leaves in Minxian, Gansu province, China. According to morphology and molecular biology properties, this virus, which has a flexuous rod-shaped particle about 750 nm in length and 12 nm in width, was assigned to the genus Potyvirus, family Potyviridae. Its coat protein (CP) shows high similarity with six other potyviruses by analysis of peptide mass fingerprinting (PMF). The 919 bp nucleotides of 3′ terminal covering partial CP gene and 3′-untranslated region was amplified by RT-PCR using degenerate primers which were designed according to the result of PMF. In sequence comparisons and phylogenetic analysis, the new isolate was found to be closely related to Japanese hornwort mosaic virus (JHMV), Konjak mosaic virus (KoMV), and Zantedeschia mosaic virus (ZaMV). The most closely related virus is JHMV03 (AB251346), with 96.59% aa and 87.60% nt identity to the isolate. All results suggest the presence of a new member of potyvirus, tentatively named Dang Gui strain of Japanese hornwort mosaic virus (JHMV-DG*). In our research the antiserum against the CP of JHMV-DG had also been prepared. To our knowledge, it is the first time that a potyvirus has been isolated and identified in Angelica sinensis. *Nucleotide sequence date reported appear in GenBank with the accession number EF157843.     

The nucleotide sequence of the 3′-terminal region of dasheen mosaic virus (Caladium isolate) and expression of its coat protein in Escherichia coli for antiserum production

Summary.  A caladium isolate of dasheen mosaic virus (DsMV-Ch) was cloned as cDNA from genomic RNA. The sequence of the 3′-terminal 3 158 nucleotides, which consisted of the 3′-terminus of the NIa gene, the NIb gene, the coat protein (CP) gene, and a 246-nucleotide non-coding region, was between 57–68% similar at the nucleotide level and 72–82% similar at the amino acid level when compared with other potyviruses. Phylogenetic analysis of aligned, selected potyviral CP sequences indicate that DsMV-Ch is similar to DsMV isolates infecting taro and closely related to the bean common mosaic virus subgroup in the genus Potyvirus. A recombinant DsMV-Ch CP (∼39 kDa) expressed in E. coli was used as an immunogen and the resulting antiserum reacted with DsMV and several other potyviruses in Western blots and indirect ELISA. Received March 19 Accepted June 26, 1998     

Characterisation of an isolate of <Emphasis Type="Italic">Narcissus degeneration virus</Emphasis> from Chinese narcissus (<Emphasis Type="Italic">Narcissus tazetta</Emphasis> var. <Emphasis Type="Italic">chinensis</Emphasis>)

Summary. A potyvirus from Chinese narcissus was transmitted mechanically to three species of Narcissus and to Lycoris radiata but not to 22 other test species. In western blot, the coat protein reacted strongly with Narcissus degeneration virus (UK isolate) antiserum. Antiserum raised to the Chinese virus did not react with eighteen other potyviruses. The complete nucleotide sequence (9816 nt) had the typical genome organisation for a member of the genus Potyvirus. Sequence comparisons and phylogenetic analysis showed that the Chinese virus was different from all previously sequenced potyviruses but distantly related to onion yellow dwarf and shallot yellow stripe viruses.     

Characterization of petunia flower mottle virus (PetFMV), a new potyvirus infecting Petunia x hybrida

Summary.  With the introduction of cutting-grown Petunia x hybrida plants on the European market, a new potyvirus which showed no serological reaction with antisera against any other potyviruses infecting petunias was discovered. Infected leaves contained flexuous rod-shaped virus particles of 750 – 800 nm in length and inclusion bodies (pinwheel structures) typical for potyviruses in ultrathin leaf sections. The purified coat protein with a M_r of approximately 36 kDa could be detected in Western immunoblots with a specific antibody to the coat protein of the petunia-infecting virus. The 3′ end of the viral genome encompassing the 3′ non-coding region, the coat protein gene, and part of the NIb gene was amplified from infected leaf material by IC/PCR using degenerate and specific primers. Sequences of PCR-generated cDNA clones were compared to other known sequences of potyviruses. Maximum homology of 56% was found in the 3′ non-coding region between the petunia isolate and other potyviruses. A maximum homology of 69% was found between the amino acid sequence of the coat protein of the petunia isolate and corresponding sequences of other potyviruses. These data indicate that the petunia-infecting virus is a previously undescribed potyvirus and the name petunia flower mottle virus (PetFMV) is suggested. Accepted November 5, 1997 Received July 25, 1997     

Nucleotide sequence of coat protein gene of yam mild mosaic virus, isolated in Papua New Guinea

Summary.  We determined the 3′-termimus 1353 nucleotides (nts) in length excluding the poly (A) tail of yam mild mosaic potyvirus (YMMV) RNA. The sequence starts within a long open reading frame (ORF) 1209 nts and is followed by untranslated region (3′-UTR) of 144 nts. The coat protein (CP) contains 266 amino acids (aa) with molecular ratio (Mr) of approximately 30 kDa. The CP of YMMV differs substantially from yam mosaic virus (YMV), Japanese yam mosaic virus (JYMV) (57 and 61% of amino acid sequence identity) and other potyvirus species. This result suggests that YMMV should be classified as a new yam potyvirus. Received January 29, 1999 Accepted February 26, 1999     

Molecular characterization and coat protein serology of watermelon leaf mottle virus (Potyvirus)

Summary.  A cDNA library was generated from purified RNA of watermelon leaf mottle virus (WLMV) (Genus Potyvirus). Two overlapping clones totaling 2,316 nucleotides at the 3′terminus of the virus were identified by immunoscreening with coat protein antiserum. The sequence analyses of the clones indicated an open reading frame (ORF) of 2,050 nucleotides which encoded part of the replicase and the coat protein, a 243-nucleotide non-coding region (3′UTR), and 23 adenine residues of the poly (A) tail. The taxonomic status of WLMV was determined by comparisons of the sequence of the cloned coat protein gene and 3′UTR with potyvirus sequences obtained from GenBank. The nucleotide sequence identities of WLMV compared with 17 other potyviruses ranged from 55.6 to 63.5% for the coat protein, and from 37.2 to 48.3% for the 3′UTR. Phylogenetic analyses of the coat protein region and the 3′UTR indicated that WLMV did not cluster with other potyviruses in a clade with high bootstrap support. The coat protein gene was expressed in Escherichia coli and a polyclonal antiserum was prepared to the expressed coat protein. In immunodiffusion tests, WLMV was found to be serologically distinct from papaya ringspot virus type W, watermelon mosaic virus 2, zucchini yellow mosaic virus, and Moroccan watermelon mosaic virus. In Western blots and ELISA, serological cross-reactivity with other cucurbit potyviruses was observed. Serological and sequence comparisons indicated that watermelon leaf mottle virus is a distinct member of the Potyvirus genus. Accepted September 23, 1999     

A new potyvirus from Thunberg fritillary (<Emphasis Type="Italic">Fritillaria thunbergii</Emphasis> Miq.) in Zhejiang,China

Summary. A potyvirus causing mosaic symptoms in Thunberg fritillary (Fritillaria thunbergii) was found at two sites in Zhejiang province, China. The virus was readily mechanically transmitted to its original host but not to any of 17 other widely used plant virus indicators. A polyclonal antiserum raised to purified virus particles reacted with its homologous virus but not with a range of other viruses (including 16 potyvirus species). In electron microscopy, virus particles and inclusion bodies typical of a potyvirus were seen. The complete nucleotide sequence of an isolate from Ningbo was determined. It was 9723 nt long and sequence analyses predicted the standard potyvirus organisation. The partial sequence (1664 nts at the 3′-terminus) of an isolate from Panan was also determined; the two sequences had 96.9% nt identity. In sequence comparisons and phylogenetic analyses with completely sequenced potyviruses, the new virus was most closely related to Lily mottle virus (53.0% aa identity) and Leek yellow stripe virus. The most closely related incomplete sequence in the international databases was for Lycoris mild mottle virus (72.8% nt identity in their coat proteins). These results suggest that the virus studied is a new species in the genus Potyvirus, which we have tentatively named Thunberg fritillary mosaic virus.     

The complete sequence of the genome of Cocksfoot streak virus (CSV), a grass infecting Potyvirus

Summary.  The complete nucleotide sequence of Cocksfoot streak virus (CSV) has been determined. The viral genome comprises 9663 nucleotides, excluding a 3′-terminal poly(A) sequence. The genome of CSV has a 133 nt 5′-non coding and a 260 nt 3′-non coding region. The RNA of CSV encodes a single polyprotein of 3089 amino acid residues and has a deduced genome organization typical for a member of the family Potyviridae. CSV is transmissible by aphids and has a narrow host range in the Gramineae. It was compared to two potyviruses having monocotyledonous hosts (monocot potyviruses) and several potyviruses infecting dicotyledonous plants (dicot potyviruses). CSV is most closely related to other monocot potyviruses like Maize dwarf mosaic virus (MDMV) and Johnson grass mosaic virus (JGMV), but also closely related to the dicot potyviruses. On the other hand, CSV is less related to monocot viruses from the other genera (Rymovirus, Tritimovirus) within the Potyviridae. Specific motifs, described for potyviral polyproteins, are also present in the polyprotein of CSV. Only two motifs in the HC-Pro – motif involved in long distance movement and motif for HC-Pro self-interaction – were different in comparison to most of the dicot potyviruses. Received September 10, 2001; accepted March 4, 2002 Published online May 24, 2002     

Phylogenetic analysis of the Potyviridae with emphasis on legume-infecting potyviruses

Summary.  The 3′-terminal nucleotide sequences of thirteen authenticated strains of bean common mosaic virus (BCMV) and one strain of bean common mosaic necrosis virus (BCMNV) were obtained. The regions sequenced included the coat protein coding sequence and 3′-end non-coding region. These data, combined with sequence information from other legume-infecting potyviruses and the Potyviridae were used for phylogenetic analysis. Evidence is provided for delineation of BCMNV as distinct from BCMV and the inclusion of azuki mosaic, dendrobium mosaic, blackeye cowpea mosaic, and peanut stripe viruses as strains of BCMV. This relationship defines the members of the BCMV and BCMNV subgroups. These data also provide a basis upon which to define virus strains, in combination with biological data. Other aspects and implications of legume-infecting potyvirus phylogenetics are discussed. Received December 24, 1996 Accepted June 3, 1997     

Molecular characterization of a distinct potyvirus from whitegrass in China

Summary.  A potyvirus isolated from perennial whitegrass (Pennisetum centrasiaticum Tzvel.) in North China was characterized at the molecular level. The 3′ terminal nucleotide (nt) sequence of 1669 nt of the viral RNA genome has been determined, which covered the coding region of the C-terminal part of the large nuclear inclusion protein (NIb, RNA polymerase), capsid protein (CP) gene and the 3′ nontranslated region (NTR). The CP gene consisted of 909 nt (including the stop codon) encoding 302 amino acid residues, and the 3′ NTR was 241 nt in length excluding the polyadenylated tract. Sequence comparison of the amino acids of CPs showed that this virus was most closely related to Sorghum mosaic virus and Maize dwarf mosaic virus with percent identities of 77% to 78% while that of the 3′ NTRs suggested that it was most closely related to Zea mosaic virus with identity of 72%. This virus isolate was to some extent closely related to other members of the Sugarcane mosaic virus subgroup of potyviruses for the CP amino acid sequences. Phylogenetic analyses of the sequences indicated that this virus isolate represented a distinct potyvirus, and the name Pennisetum mosaic virus (PenMV) is proposed. Received November 22, 2002; accepted January 8, 2003 Published online March 21, 2003     

Detection and characterisation of a second potyvirus from Thunberg fritillary in China

Summary. Plants of Thunberg fritillary (Fritillaria thunbergii Miq.) from Zhejiang Province, were found to be co-infected with two distinct potyviruses. One was an isolate of the recently reported Thunberg fritillary mosaic virus (TFMV; Wei et al., (2005) Arch Virol 150: 1271–1280), while the other was a distinct virus that did not react with TFMV antiserum nor with antisera to 17 other potyviruses, except for a weak reaction with antibodies produced to soybean mosaic virus (SMV) Pinellia strain. Both viruses could be transmitted mechanically to their original host but not to any of a range of commonly used indicator plants. No local lesion host was identified that would enable the viruses to be propagated independently. The complete sequences of both viruses were determined; that of the new virus (9656 nt) had the typical genome organisation and recognised sequence motifs of a potyvirus, encoding a putative polyprotein of 351 kDa. Phylogenetic analysis, sequence comparisons, and the pattern of polyprotein cleavage sites all indicated that it was a member of the Bean common mosaic virus subgroup. The most closely related species are Soybean mosaic virus and Wisteria vein mosaic virus, with 68–69% amino acid identity between their polyproteins. This is sufficiently different for the new virus to be regarded as a distinct species, which we have tentatively named Fritillary virus Y.     

Pepper yellow mosaic virus,a new potyvirus in sweetpepper,Capsicum annuum

Summary.  A potyvirus was found causing yellow mosaic and veinal banding in sweetpepper in Central and Southeast Brazil. The sequence analysis of the 3′ terminal region of the viral RNA revealed a coat protein of 278 amino acids, followed by 275 nucleotides in the 3′-untranslated region preceding a polyadenylated tail. The virus shared 77.4% coat protein amino acid identity with Pepper severe mosaic virus, the closest Potyvirus species. The 3′-untranslated region was highly divergent from other potyviruses. Based on these results, the virus found in sweetpepper plants could be considered as a new potyvirus. The name Pepper yellow mosaic virus (PepYMV) is suggested. Received March 12, 2001 Accepted August 28, 2001     

Identification and molecular characterization of a new potyvirus infecting Triteleia species

Plants of Triteleia hyacinthina, Triteleia ixioides Starlight, and Triteleia laxa Corina with severe mosaic and yellow vein-banding were found to be infected with a potyvirus. The 3′-terminal region of the virus was amplified by RT-PCR from total RNA using a potyvirus-specific degenerate primer (poty5P: 5′ GGN AAY AAY AGY GGN CAR CC 3′) and an oligo-dTprimer. The sequence generated included the 3′-NIb protein coding region (680 nucleotides), the entire coat protein coding region (840 nucleotides), and 3’-untranslated region (UTR) (253 nucleotides). Amino acid identity of the whole CP between the triteleia virus and potyvirus member ranged from 54% Apium virus Y (ApVY) to 67% Auraujia mosaic virus (ArjMV) and Twisted-stalk chlorotic streak virus (TSCSV) and the core ranged from 59% (ApVY) to 75% (ArjMV). The 3-UTR showed no significant homology with other known potyviruses. Phylogenetic relationships suggest this triteleia virus is a new member of the Potyvirus genus and the name of “Triteleia mosaic virus” (TriMV) is proposed. This is the first report of a potyvirus infecting triteleia.     

Identification and full sequence of an isolate of Alternanthera mosaic potexvirus infecting Phlox stolonifera

Summary. A potexvirus was isolated from creeping phlox (Phlox stolonifera) plants from a commercial nursery in Pennsylvania. The virus was serologically related to clover yellow mosaic virus, plantain virus X, potato virus X, and potato aucuba mosaic virus, and was most closely related to papaya mosaic virus (PapMV). The sequence of a PCR fragment obtained with potexvirus group-specific primers was distinct from that of PapMV; the coat protein (CP) gene and 3′ untranslated region (UTR) were closely related to Alternanthera mosaic virus (AltMV), previously reported only from Australia. The host range was similar to that of the Australian isolate (AltMV-Au), and the phlox isolate reacted strongly with antiserum to AltMV-Au. The full sequence of the phlox isolate was more closely related to PapMV throughout the genome than to any potexvirus other than AltMV-Au, for which only the CP and 3′UTR sequences are available. The phlox isolate was therefore named AltMV-PA (for Pennsylvania), and the full 6607 nt sequence is presented¹. Additional AltMV isolates from creeping phlox (AltMV-BR and AltMV-SP) and trailing portulaca (Portulaca grandiflora; AltMV-Po) were also isolated, suggesting that AltMV may be widespread, and may have been mis-diagnosed in the past as PapMV. AltMV has the potential to spread to other ornamental crops. ¹The full sequence of AltMV-PA has been deposited in GenBank as Accession Number AY863024; the 3′-terminal sequences of AltMV-PA, AltMV-BR, AltMV-SP, and AltMV-Po have been deposited under Accession numbers AY850929, AY850928, AY850931, and AY850930, respectively.