First report of bean yellow mosaic virus infecting Gladiolus sp. in Ukraine
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First report of bean yellow mosaic virus infecting Gladiolus sp. in Ukraine Lidiya Mishchenko 1 Hanna Korotieieva 1
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Alina Dunich 1
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Andrei Smertenko 2
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Anna Dashchenko 3 & Roksolana Sovinska 1 &
Received: 20 November 2019 / Accepted: 25 January 2020 # Società Italiana di Patologia Vegetale (S.I.Pa.V.) 2020
Keywords Bean yellow mosaic virus . Gladiolus . Potyvirus . Ukraine
Bean yellow mosaic virus (BYMV) is the one of most prevalent and harmful viruses infecting gladiolus plants worldwide. BYMV has previously been identified on lupin, soybean, and bean in central Ukraine (Kyrychenko et al. 2017). Several cases of gladiolus infection by BYMV were already reported in different countries; however no cases were reported in the Ukraine thus far. From 2009 to 2018, we found Gladiolus sp. plants showing typical symptoms of viral disease including leaf chlorotic stripes, mosaics, and delay in flowering. Symptoms were identical every year of observations and revealed on the same cultivar ‘Pamyat`’ in Poltava and Kyiv regions. Electron microscopic investigations of sap from 50 gladiolus samples showed filamentous viral particles 750 ± 20 by 12 nm typical for potyviruses. To identify the virus in gladiolus plants, we performed double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) using specific antibodies against BYMV (Loewe, Germany). We also used two step RTPCR with BYMV-specific oligonucleotide primers for
* Lidiya Mishchenko [email protected] 1
ESC “Institute of Biology and Medicine”, Taras Shevchenko National University of Kyiv, Volodymyrska Street, 64/13, Kyiv 01601, Ukraine
2
Institute of Biological Chemistry, Washington State University, PO Box 646340, Pullman, WA 99164, USA
3
National University of Life and Environmental Sciences of Ukraine, Heroyiv Oborony Street, 15, Kyiv 03041, Ukraine
amplifying the 590 bp fragment of BYMV coat protein gene (Wang et al. 2017). Total RNA was extracted from fresh leaves using RNeasy Plant Mini kit (Qiagen, Germany) following the manufacturer’s instructions. The cDNA was synthetized using RevertAid Reverse Transcriptase (MMuLV RT; Thermo Scientific, USA) according to the manufacturer’s instructions. ELISA data showed the presence of BYMV in all of 120 total symptomatic gladiolus leaf samples collected from 2009 to 2018. Fifty ELISA positive symptomatic leaf samples were tested with RT-PCR, which confirmed BYMV presence in all samples. Partial sequences (578 bp) of the coat protein gene of two Ukrainian BYMV isolates GlMP-17 and GlMP-18 collected in 2017 and 2018 in Poltava region were submitted to the NCBI GenBank under accession numbers MK416161 and MK416160, respectively. The highest amino acid (aa; 96.9–100%) and nucleotide (nt; 96.5–98.6%) sequence identity of the Ukrainian isolates GlMP-17 and GlMP-18 was found to be with Japanese isolate 35–1 from Gentiana plants (AB097090) and Chinese isolate Saffron2 from Crocus sativus plants (MG002647). The nt and aa sequences corresponding to 578 nt of the coat protein located at the po
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