[Show abstract][Hide abstract] ABSTRACT: The complete genomic RNA sequences of the tymovirus isolates Hu and Col from potato which originally had been considered to be strains of the same virus species, i.e. Andean potato latent virus (APLV), were determined by siRNA sequencing and assembly, and found to share only c. 65% nt sequence identity. This result together with those of serological tests and comparisons of the coat protein gene sequences of additional tymovirus isolates from potato suggest that the species Andean potato latent virus should be subdivided into two species, i.e. APLV and Andean potato mild mosaic virus (APMMV). Primers were designed for the broad specificity detection of both viruses.
[Show abstract][Hide abstract] ABSTRACT: Virus infection of Lagenaria breviflora and Coccinia barteri were observed in Calabar, Nigeria during the 2005/2006 growing season. The viruses causing the diseases were characterized in this study. Diagnostic tools were host range, transmission studies, cytopathology, electron microscopy, Immunosorbent Electron Microscopy (ISEM), serology and coat protein gene sequencing. Evidence from biological, serological and sequence data confirmed that both viruses belong to the genus Potyvirus, family Potyviridae. Both were mechanically transmissible and also transmitted by Myzus persicae and Aphis gossyppi in a fore-gut manner. They also induced cytoplasmic inclusions in infected leaf tissues. The Lagenaria virus isolate has a coat protein molecular weight of 32.5 kDa and 35.0 kDa for Coccinia isolate. Of the two viruses only Lagenaria virus isolate showed cross reactivity with MWMV in DAS-ELISA. Both viruses reacted negatively with antisera to some notable cucurbit viruses in the same test, showed weak to moderate decorations with antisera to PRSV, TuMV and TeMV in immunosorbent electron microscopy (ISEM) tests. The Coccinia virus was, however, strongly decorated by antiserum to MWMV but no decoration with the Lagenaria virus. Comparison of the amino acid sequence data of the N-terminal regions of the coat proteins to that of MWMV reported from Sudan indicated 92% and 93% identities for the Coccinia and Lagenaria viruses, respectively. It is suggested that the virus isolates reported in the study be considered strains of the MWMV Sudan isolate. This is the first report of the occurrence of MWMV in Nigeria.
Full-text · Article · Mar 2012 · International Journal of Virology
[Show abstract][Hide abstract] ABSTRACT: The complete genome of a tomato mild mottle virus (ToMMV) isolate was analysed, and some biological features were characterized. The ssRNA genome of ToMMV from Ethiopia encompasses 9283 nucleotides (excluding the 3' poly(A) tail) and encodes a polyprotein of 3011 amino acids. Phylogenetic and pairwise comparisons with other members of the family Potyviridae revealed that ToMMV is the most divergent member of the genus Ipomovirus, with a genome organization similar to that of members of the species Sweet potato mild mottle virus, the type species of the genus. In contrast to earlier reports, ToMMV isolates from Yemen and Ethiopia were not transmitted by the aphid Myzus persicae, but they were transmitted very erratically by the whitefly Bemisia tabaci. A comparison of the 3'-proximal sequences of different isolates provided evidence for geographically associated genetic variation.
No preview · Article · Nov 2011 · Archives of Virology
[Show abstract][Hide abstract] ABSTRACT: Chickpea plants with severe yellowing and tip wilting were observed in the Cap-Bon Region of Tunisia in 2006. The viral-like symptoms resulted in yield loss of approximately 25% in some fields. A total of 110 symptomatic chickpea plants was collected from nine chickpea fields and tested at the Virology Laboratory of ICARDA, Syria for eight legume viruses using tissue-blot immunoassay (TBIA) (3). Polyclonal antisera produced at the ICARDA Virology Laboratory were used to test for Chickpea chlorotic dwarf virus (genus Mastrevirus, family Geminiviridae), Broad bean stain virus (genus Comovirus, family Secoviridae), Broad bean mottle virus (genus Bromovirus, family Bromoviridae), and Bean yellow mosaic virus and Pea seed borne mosaic virus (genus Potyvirus, family Potyviridae). Antiserum to Beet mosaic virus (BtMV; genus Potyvirus, family Potyviridae) (AS-0143) was provided by the German Collection of Microorganisms and Cell Cultures (DSMZ, Braunschweig, Germany). In addition, three monoclonal antibodies (MAb) were used to detect Faba bean necrotic yellows virus (FBNYV; genus Nanovirus, family Nanoviridae) (MAb 3-2E9) (1), potyviruses (PVAS-769 [MAb PTY 3 Potyvirus Group] American Type Culture Collection, Manassas, VA), and luteoviruses (MAb B-2-5G4) (2). Twenty-two of the plants tested positive with MAb PTY 3 and BtMV antisera, 56 samples reacted with MAb B-2-5G4, and eight plants with the FBNYV MAb, whereas 24 plants tested negative with all antisera. Because reactions with the BtMV antiserum were unexpected, detection of BtMV was confirmed by reverse transcription-(RT)-PCR assays using BtMV-specific primers (LN26 and LN27) (4), which produced an amplicon of expected size (1,050 bp) from all plants that reacted with BtMV antiserum but not from plants that were serologically negative. Leaf tissue from a BtMV-infected plant was ground in 0.01 M potassium phosphate buffer, pH 7.2 (1:20, wt/vol), mixed with 0.5% celite, and used for mechanical inoculation of chickpea seedlings (cv. Beja 4). In addition, adults of three legume aphid species (Aphis craccivora, A. fabae, and Acyrthosiphon pisum) were starved for 1 h before feeding on BtMV-infected chickpea leaves for an acquisition access period of 5 min. Fifteen aphids of each species were placed on each chickpea plant, allowed to feed for 24 h, and then sprayed with an insecticide. Tip wilting symptoms appeared on plants 15 to 20 days after mechanical and aphid inoculations but not on plants used as negative control treatments (inoculated mechanically with healthy leaf tissue or with aphids that had fed on noninfected chickpea plants). Use of BtMV antiserum for TBIA analysis of inoculated plants revealed systemic BtMV infections in 35 of 92 plants inoculated mechanically and 15 of 75 plants inoculated with viruliferous A. fabae only. To our knowledge, this is the first record of BtMV infecting chickpea in Tunisia.
References: (1) A. Franz et al. Ann. Appl. Biol. 128:255, 1996. (2) L. Katul. Characterization by serology and molecular biology of bean leaf roll virus and faba bean necrotic yellows virus. Ph.D. thesis. University of Gottingen, Germany, 1992. (3) K. M. Makkouk and A. Comeau. Eur. J. Plant Pathol. 100:71, 1994. (4) L. G. Nemchinov et al. Arch. Virol. 149:1201, 2004.
[Show abstract][Hide abstract] ABSTRACT: Using monoclonal antibodies raised against a Faba bean necrotic yellows virus (FBNYV) isolate from Egypt and a Faba bean necrotic stunt virus (FBNSV) isolate from Ethiopia, a striking serological variability among nanovirus isolates from faba bean in Morocco was revealed. To obtain a better understanding of this nanovirus variability in Morocco, the entire genomes of two serologically contrasting isolates referred to as Mor5 and Mor23 were sequenced. The eight circular ssDNA components, each identified from Mor5- and Mor23-infected tissues and thought to form the complete nanovirus genome, ranged in size from 952 to 1,005 nt for Mor5 and from 980 to 1,004 nt for Mor23 and were structurally similar to previously described nanovirus DNAs. However, Mor5 and Mor23 differed from each other in overall nucleotide and amino acid sequences by 25 and 26%, respectively. Mor23 was most closely related to typical FBNYV isolates described earlier from Egypt and Syria, with which it shared a mean amino acid sequence identity of about 94%. On the other hand, Mor5 most closely resembled a FBNSV isolate from Ethiopia, with which it shared a mean amino acid sequence identity of approximately 89%. The serological and genetic differences observed for Mor5 and Mor23 were comparable to those observed earlier for FBNYV, FBNSV, and Milk vetch dwarf virus. Following the guidelines on nanovirus species demarcation, this suggests that Mor23 and Mor5 represent isolates of FBNYV and FBNSV, respectively. This is the first report not only on the presence of FBNSV in a country other than Ethiopia but also on the occurrence and complete genome sequences of members of two nanovirus species in the same country, thus providing evidence for faba bean crops being infected by members of two distinct nanovirus species in a restricted geographic area.
Preview · Article · Jan 2010 · Archives of Virology
[Show abstract][Hide abstract] ABSTRACT: Some legume samples with yellowing and stunting symptoms from Ethiopia and Sudan that serologically reacted with a broad-spectrum luteovirus monoclonal antibody did not react or very weakly reacted with virus-specific antibodies suggesting the occurrence of new luteovirus variants. Reverse transcriptase (RT)-PCR amplification, cloning, nucleotide sequencing and analysis of coat protein (CP) gene of a luteovirus isolate from chickpea in Sudan indicated that it shares a closest predicted amino acid sequence identity of only 66% with Soybean dwarf virus (SbDV). Since this is less than the accepted threshold value of 90% recommended for discriminating luteovirus species, the isolate is suggested to represent a distinct luteovirus for which the name Chickpea yellows virus (CpYV) is proposed. Similarly, a lentil isolate from Ethiopia shared a closest CP amino acid sequence identity of 86% with viruses of the Beet western yellows virus subgroup. Following the same criteria, this isolate represents another distinct luteovirus species for which the name Lentil stunt virus (LStV) is suggested. From faba bean, CP sequences of Turnip yellows virus were amplified from Egyptian and Moroccan samples whereas partial CP sequences of SbDV were amplified from Ethiopian, Syrian and Chinese samples. The study indicated that legume luteoviruses in northeast Africa are highly diverse.
Full-text · Article · Jan 2010 · AFRICAN JOURNAL OF BIOTECHNOLOGY
[Show abstract][Hide abstract] ABSTRACT: A high-molecular-weight dsRNA (approximately 15 kbp) was isolated from chlorotic leaves of a carrot plant and used for determining the entire nucleotide sequence of a closterovirus. The complete genome of this carrot closterovirus (CCV) was 16.4 kb in length and contained ten open reading frames (ORFs). The genome organization of CCV resembled that of beet yellow stunt virus, but ORF2 and ORF3 were in a reversed order. Based on Hsp70h sequences, CCV is most closely related to carnation necrotic fleck virus and mint virus 1, two viruses of the genus Closterovirus (family Closteroviridae). The major coat protein gene of CCV was expressed in Escherichia coli for raising an antiserum. This permitted routine detection of CYLV by DAS-ELISA and immunoelectron microscopy and was used for demonstrating the bipolar nature of the CCV virion. Moreover, the antiserum gave a Western blot reaction with a reference sample of a Carrot yellow leaf virus (CYLV) isolate from the Netherlands, suggesting that CCV is a German isolate of CYLV.
No preview · Article · Aug 2009 · Archives of Virology
[Show abstract][Hide abstract] ABSTRACT: Chickpea chlorotic stunt virus (CpCSV), a proposed new member of the genus Polerovirus (family Luteoviridae), has been reported only from Ethiopia. In attempts to determine the geographical distribution and variability of CpCSV, a pair of degenerate primers derived from conserved domains of the luteovirus coat protein (CP) gene was used for RT-PCR analysis of various legume samples originating from five countries and containing unidentified luteoviruses. Sequencing of the amplicons provided evidence for the occurrence of CpCSV also in Egypt, Morocco, Sudan, and Syria. Phylogenetic analysis of the CP nucleotide sequences of 18 samples from the five countries revealed the existence of two geographic groups of CpCSV isolates differing in CP sequences by 8-10%. Group I included isolates from Ethiopia and Sudan, while group II comprised those from Egypt, Morocco and Syria. For distinguishing these two groups, a simple RFLP test using HindIII and/or PvuII for cleavage of CP-gene-derived PCR products was developed. In ELISA and immunoelectron microscopy, however, isolates from these two groups could not be distinguished with rabbit antisera raised against a group-I isolate from Ethiopia (CpCSV-Eth) and a group-II isolate from Syria (CpCSV-Sy). Since none of the ten monoclonal antibodies (MAbs) that had been produced earlier against CpCSV-Eth reacted with group-II isolates, further MAbs were produced. Of the seven MAbs raised against CpCSV-Sy, two reacted only with CpCSV-Sy and two others with both CpCSV-Sy and -Eth. This indicated that there are group I- and II-specific and common (species-specific) epitopes on the CpCSV CP and that the corresponding MAbs are suitable for specific detection and discrimination of CpCSV isolates. Moreover, CpCSV-Sy (group II) caused more severe stunting and yellowing in faba bean than CpCSV-Eth (group I). In conclusion, our data indicate the existence of a geographically associated variation in the molecular, serological and presumably biological properties of CpCSV.
Full-text · Article · May 2009 · Archives of Virology
[Show abstract][Hide abstract] ABSTRACT: A virus disease of faba bean (Vicia faba L.) in China, characterized by leaf yellowing and rolling and plant stunting, was shown to be caused by a virus of the genus Nanovirus based on serological reactions to nanovirus-specific monoclonal antibodies and the generation of polymerase chain reaction amplicons using nanovirus-specific primers. To identify the faba bean-infecting nanovirus, regions of the DNA components encoding the master replication initiator protein and capsid protein of two nanovirus isolates from China were cloned, sequenced and compared with those of other members of the genus Nanovirus. The two Chinese virus isolates shared nucleotide sequence identities ranging from 95 to 98% with the type isolate of Milk vetch dwarf virus (MDV) from Japan. They were thus identified as isolates of MDV, a virus so far known to cause important diseases of legumes in Japan. This is the first record of MDV-infecting faba bean in China.
No preview · Article · Apr 2009 · Journal of Phytopathology
[Show abstract][Hide abstract] ABSTRACT: Sweet potato chlorotic fleck virus (SPCFV) has recently been classified as it putative new member of the genus Carlavirus (family Flexiviridae) on the basis of its Molecular properties. In this study, SPCFV was characterized in terms of host range, physical and biological characteristics, and genetic variability. In addition to sweet potato. SPCFV infected sonic plant species in the families Convolvulaceae, Chenopodiaceae, and Solanaceae. Limited numbers of virus particles were observed in the assimilation parenchyma cells of infected plant tissues. sonic cells had it distorted and enlarged endoplasmic reticulum though without any cytoplasmic and amorphous inclusions. The normal length of SPCFV particles was determined to he approximately 800 nm, In enzyme-linked immunosorbent assays, polyclonal antibodies raised against purified SPCFV virions were able to detect the virus in infected sweet potato and indicator plant tissues. In immunoelectron microscopy, SPCFV particles were all strongly decorated when reacted with homologous antiserum. Comparison of the 3' terminal part of the genome of a range of geographically diverse isolates revealed a high level of genetic diversity. The amino acid Sequence identity in the coat protein and the nucleic acid binding protein ranged front 89 to 99.7% and front 75.9 to 99.2%, respectively. Phylogenetic analysis of both proteins showed a geographically associated Clustering into two genogroups.
[Show abstract][Hide abstract] ABSTRACT: Carrot motley dwarf (CMD) is known to result from a mixed infection by two viruses, the polerovirus Carrot red leaf virus and one of the umbraviruses Carrot mottle mimic virus or Carrot mottle virus. Some umbraviruses have been shown to be associated with small satellite (sat) RNAs, but none have been reported for the latter two. A CMD-affected parsley plant was used for sap transmission to test plants, that were used for dsRNA isolation. The presence of a 0.8-kbp dsRNA indicated the occurrence of a hitherto unrecognized satRNA associated with CMD. The satRNAs of the CMD isolate from parsley and an isolate from carrot have been sequenced and showed 94% sequence identity. Nucleotide sequences and putative translation products had no significant similarities to GenBank entries. To our knowledge, this is the first report of satRNAs associated with CMD.
[Show abstract][Hide abstract] ABSTRACT: The effect of sowing date on aphid infestation and the incidence of aphid-transmitted viruses were investigated in organically managed, small-scale field experiments with two faba bean cultivars over 3 years (2002–04). As an additional factor, straw mulch was applied in 2 of the 3 years shortly before the start of vector activity in May. Virus incidence was determined using enzyme-linked immunosorbent assay and immunoelectron microscopy. Aphid flight activity was monitored using standard yellow water traps. Bean colonising aphids were assessed throughout the vegetation period by counting the number of plants infested with Acyrthosiphon pisum, Megoura viciae and Aphis fabae. Pea enation mosaic virus and bean yellow mosaic virus were the most abundant aphid-transmitted viruses, being detected in 22–54% and 9–69%, respectively, of the total number of virus-infected plants analysed per year. Further aphid-transmitted viruses found in faba bean were bean leaf roll virus, beet western yellows virus, clover yellow vein virus (in 2002) and soybean dwarf virus (in 2004). A. pisum was the predominant aphid species colonising faba bean plants. Early sowing compared with late sowing led to a significant reduction of the total virus incidence in faba bean in all 3 years. However, significantly decreased levels of A. pisum colonisation as a result of early sowing were observed only in 1 year and one cultivar. Irrespective of sowing date, straw mulching had no significant effects on virus incidence and aphid colonisation. Compared with late sowing, early sowing significantly increased bean yield in all 3 years and kernel weight in 2 years, whereas straw mulching had no effect on yield.
Full-text · Article · Oct 2008 · Annals of Applied Biology