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First Report of Cucurbit Aphid-Borne Yellows Virus Infecting Zucchini Plants (Cucurbita pepo convar. giromontiina) in Poland

Zarzyńska-Nowak, A., Hasiów-Jaroszewska, B., Budzyńska, D., Borodynko-Filas, N.
Plant disease 2019 v.103 no.5 pp. 1047
Aphis gossypii, Cucumber mosaic virus, Cucurbit aphid-borne yellows virus, Cucurbita pepo, Myzus persicae, Papaya ringspot virus, RNA, Watermelon mosaic virus, Zucchini yellow mosaic virus, abnormal development, antiserum, chlorosis, coat proteins, control methods, crop production, crops, disease surveillance, enzyme-linked immunosorbent assay, fruits, genes, genetic vectors, growth retardation, mixed infection, necrosis, nucleotide sequences, oligodeoxyribonucleotides, plant diseases and disorders, plant viruses, plasmids, sequence analysis, surveys, vector control, viruses, zucchini, Czech Republic, France, Montenegro, Poland
In 2018, surveys were conducted in the major cucurbit-growing areas in western Poland to assess the prevalence and distribution of viral diseases. In all of the investigated fields, the presence of aphids has been previously observed. A total of 68 symptomatic zucchini (Cucurbita pepo convar. giromontiina) plant samples showing virus-like symptoms, such as vein clearing, severe mosaic, mottling, chlorosis, necrosis, stunting, fruit discoloration, and malformations, and 11 asymptomatic samples were collected from 18 fields in Wielkopolska and Kujawsko-Pomorskie regions. Virus identification was carried out using an enzyme-linked immunosorbent assay using antisera raised against cucurbit aphid-borne yellows virus (CABYV, genus Polerovirus, family Luteoviridae), cucumber mosaic virus (CMV), papaya ringspot virus (PRSV), watermelon mosaic virus (WMV), and zucchini yellow mosaic virus (ZYMV) (DSMZ, Braunschweig, Germany). Out of the 79 tested samples, 52 were positive for ZYMV, 31 for WMV, 12 for CMV, and 9 for CABYV in both single and multiple infections. The presence of PRSV was not confirmed in the analyzed samples. Moreover, the presence of the abovementioned viruses was not confirmed in asymptomatic samples. CABYV isolates were detected in two separate fields in the Wielkopolska region in single (one sample) and mixed infection with WMV (three samples) and ZYMV (one sample). In four samples, the mixed infection of CABYV, WMV, and ZYMV was identified. To confirm the presence of CABYV, total RNA was extracted from nine samples using the RNeasy Plant Mini Kit (Qiagen, Hilde, Germany) and subsequently used as templates in reverse transcription polymerase chain reactions (RT-PCRs) with a specific primer set: CABYV-CP-F/CABYV-CP-Rev amplifying the full length of coat protein (cp) (Choi et al. 2015). The expected PCR products of approximately 600 bp were observed on 1% agarose gel for all of the tested samples. Subsequently, two randomly selected RT-PCR products were ligated into the TOPO TA Cloning Vector for Sequencing (Life Technologies, Waltham, MA), and plasmid DNA was sequenced by an external company (Genomed, Warsaw, Poland). The obtained cp gene sequences revealed 100% identity; therefore, one representative sequence was deposited in GenBank under accession number MK059479. The comparison of the cp gene of the Polish CABYV isolates with the others described to date revealed 95 to 99% nucleotide sequence identity. The highest nucleotide sequence identity the Polish isolates shared with isolates collected from, for example, Montenegro, the Czech Republic, and France (KX398664, KX398665, HM771269, and X76931). To our knowledge, this is the first report of the occurrence of CABYV in Poland. The virus is transmitted in a persistent manner by several aphis species such as Aphis gossypii and Myzus persicae, which are both common in Poland; therefore, the potential virus spreading is a serious threat to zucchini production in Poland. Further disease monitoring and the establishment of effective vector control methods will be needed for disease management in zucchini as well as in other cucurbit crops in Poland.