Characterization of a Syrian Chickpea chlorotic stunt virus strain and production of polyclonal antibodies for its detection

Reverse transcription-polymerase chain reaction analysis with two primer sets of luteoviruses was used to characterize an isolate of Chickpea chlorotic stunt virus (CpCSv, genus Polerovirus, family Luteoviridae) (SC402-08) collected from Lattakia, Syria, during the 2007‒2008 chickpea growing season. Sequence analysis revealed that the coat protein gene of the isolate shared nucleotide sequence identities ranging from 97 to 98% with the CpCSv isolates from Egypt, morocco and Syria. The capsid protein was separated as a protein of approximately 20 kDa in sodium dodecyl sulphate polyacrylamide gel electrophoresis, and was visually detected by its reaction with CpCSV monoclonal antibody in Western blot. SC402-08 isolate of CpCSV was purified from faba bean-infected plants, and yielded 112–182 μg of purified virions kg-1 of infected tissue. The purified preparation was injected into a white rabbit, and an antiserum was obtained and used to detect CpCSv in infected tissues by tissue-blot immunoassay. The antiserum obtained was able to detect CpCSv by the immunoassay up to a dilution of 1:1,024,000

Characterization of a Syrian <em>Chickpea chlorotic stunt virus</em> strain and production of polyclonal antibodies for its detection

Reverse transcription-polymerase chain reaction analysis with two primer sets of luteoviruses was used to characterize an isolate of <em>Chickpea chlorotic stunt virus</em> (CpCSv, genus <em>Polerovirus</em>, family <em>Luteoviridae</em>) (SC402-08) collected from Lattakia, Syria, during the 2007‒2008 chickpea growing season. Sequence analysis revealed that the coat protein gene of the isolate shared nucleotide sequence identities ranging from 97 to 98% with the CpCSv isolates from Egypt, morocco and Syria. The capsid protein was separated as a protein of approximately 20 kDa in sodium dodecyl sulphate polyacrylamide gel electrophoresis, and was visually detected by its reaction with CpCSV monoclonal antibody in Western blot. SC402-08 isolate of CpCSV was purified from faba bean-infected plants, and yielded 112–182 μg of purified virions kg-1 of infected tissue. The purified preparation was injected into a white rabbit, and an antiserum was obtained and used to detect CpCSv in infected tissues by tissue-blot immunoassay. The antiserum obtained was able to detect CpCSv by the immunoassay up to a dilution of 1:1,024,000

Hidden diversity of Macrophomina associated with broadacre and horticultural crops in Australia

Worldwide, most isolates of Macrophomina (Botryosphaeriaceae) have been attributed to the generalist phytopathogen M. phaseolina. Since 2014, three cryptic species of Macrophomina have been recognised by molecular methods. This study elucidates the taxonomy of Macrophomina species associated with broadacre and horticultural crops in Australia. A five-locus phylogenetic analysis of 80 isolates of Macrophomina from 28 plant species in Australia, combined with genealogical concordance phylogenetic species recognition and coalescent-based species delimitation approaches, found M. phaseolina, M. pseudophaseolina and supported the introduction of M. tecta sp. nov. Macrophomina phaseolina was the most frequently isolated (88%). Macrophomina pseudophaseolina is reported for the first time in Australia. Macrophomina tecta was isolated from stems of Sorghum bicolor and Vigna radiata with charcoal rot symptoms in New South Wales and Queensland. The potential for two or more Macrophomina species to co-infect the same host has implications for disease epidemiology and pathogen evolution. Future investigations into the distribution, biology, host range and population diversity of the new Macrophomina records are needed

Hidden diversity of Macrophomina associated with broadacre and horticultural crops in Australia

Worldwide, most isolates of Macrophomina (Botryosphaeriaceae) have been attributed to the generalist phytopathogen M. phaseolina. Since 2014, three cryptic species of Macrophomina have been recognised by molecular methods. This study elucidates the taxonomy of Macrophomina species associated with broadacre and horticultural crops in Australia. A five-locus phylogenetic analysis of 80 isolates of Macrophomina from 28 plant species in Australia, combined with genealogical concordance phylogenetic species recognition and coalescent-based species delimitation approaches, found M. phaseolina, M. pseudophaseolina and supported the introduction of M. tecta sp. nov. Macrophomina phaseolina was the most frequently isolated (88%). Macrophomina pseudophaseolina is reported for the first time in Australia. Macrophomina tecta was isolated from stems of Sorghum bicolor and Vigna radiata with charcoal rot symptoms in New South Wales and Queensland. The potential for two or more Macrophomina species to co-infect the same host has implications for disease epidemiology and pathogen evolution. Future investigations into the distribution, biology, host range and population diversity of the new Macrophomina records are needed