Molecular,serological and biological variation among chickpea chlorotic stunt virus isolates from five countries of North Africa and West Asia |
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Authors: | A D Abraham W Menzel M Varrelmann and H Josef Vetten |
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Institution: | 1.Julius Kuehn Institute, Federal Research Center for Cultivated Plants (JKI),Braunschweig,Germany;2.Section Plant Virology, Department of Crop Sciences,University of G?ttingen,G?ttingen,Germany;3.Biotechnology Program,Ethiopian Institute of Agricultural Research,Addis Ababa,Ethiopia |
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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. |
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