ena-STUDY-Aristotle University of Thessaloniki-04-04-2018-11:51:30:873-3283
First report of Capsicum chlorosis virus in pepper in Greece
In November 2017, virus-like symptoms similar to those caused by Tospoviruses were observed in a greenhouse pepper crop (cultivar Areti) in Ierapetra, Crete. The symptoms included necrotic ringspots, mottling of leaves and appeared in low incidence of approximately 1%. In total, samples were analyzed both serologically and by TSWV-specific primers TSWVF/TSWVR (Hu et al., 2018)tested for the presence of Tomato spotted wilt virus (TSWV), a virus endemic in Greece for more 50 years. As both methods gave negative results, the samples were also tested by a generic RT-PCR assay against Tospoviruses using the primer sets gM410/gM870c (Chen et al., 2012). A DNA band of 500 bp corresponding to NSm gene (Chen et al., 2012) was amplified in all symptomatic samples. Two of the produced amplicons were directly sequenced and further nucleotide comparison revealed that the obtained sequences were 98% identical to an isolate of Capsicum chlorosis virus (CaCV) from India (Accession No. KX499515). Therefore, the samples were also tested with the primers CaCF/CaCR for specific detection of a DNA fragment of approximately 850 bp corresponding to the nucleocapsid (N) gene of CaCV (Krishnareddy et al., 2008). The RT-PCR reaction confirmed the presence of the virus in all tested samples and subsequent sequencing analysis of the two previous positive isolates revealed their 99% nucleotide identity with an isolate from India (EU216023). Mechanical inoculation was used for further confirmation of the virus. Symptomatic leaves were inoculated onto 10 Nicotiana benthamiana indicator plants. Seven to 10 days post inoculation, wilt??? symptoms started to appear on N. benthamiana plants, which are consistent of tospovirus infection. RT-PCR assay confirmed the presence of CaCV in_/10 inoculated plants. So far, CaCV has only been reported in Australia and South-East Asia. To our knowledge, this is the first report of CaCV in Greece and in the Mediterranean region. Extensive surveys in other regions are required in order to unravel the spread of this new pathogen in pepper and other crops in Greece.
In November 2017, virus-like symptoms similar to those caused by Tospoviruses were observed in a greenhouse pepper crop (cultivar Areti) in Ierapetra, Crete. The symptoms included necrotic ringspots, mottling of leaves and appeared in low incidence of approximately 1%. In total, samples were analyzed both serologically and by TSWV-specific primers TSWVF/TSWVR (Hu et al., 2018)tested for the presence of Tomato spotted wilt virus (TSWV), a virus endemic in Greece for more 50 years. As both methods gave negative results, the samples were also tested by a generic RT-PCR assay against Tospoviruses using the primer sets gM410/gM870c (Chen et al., 2012). A DNA band of 500 bp corresponding to NSm gene (Chen et al., 2012) was amplified in all symptomatic samples. Two of the produced amplicons were directly sequenced and further nucleotide comparison revealed that the obtained sequences were 98% identical to an isolate of Capsicum chlorosis virus (CaCV) from India (Accession No. KX499515). Therefore, the samples were also tested with the primers CaCF/CaCR for specific detection of a DNA fragment of approximately 850 bp corresponding to the nucleocapsid (N) gene of CaCV (Krishnareddy et al., 2008). The RT-PCR reaction confirmed the presence of the virus in all tested samples and subsequent sequencing analysis of the two previous positive isolates revealed their 99% nucleotide identity with an isolate from India (EU216023). Mechanical inoculation was used for further confirmation of the virus. Symptomatic leaves were inoculated onto 10 Nicotiana benthamiana indicator plants. Seven to 10 days post inoculation, wilt??? symptoms started to appear on N. benthamiana plants, which are consistent of tospovirus infection. RT-PCR assay confirmed the presence of CaCV in_/10 inoculated plants. So far, CaCV has only been reported in Australia and South-East Asia. To our knowledge, this is the first report of CaCV in Greece and in the Mediterranean region. Extensive surveys in other regions are required in order to unravel the spread of this new pathogen in pepper and other crops in Greece.