Glycine max (Soybean) is the most important edible crop in Korea. In Korea, eight viruses have been reported to infect soybean, including Alfalfa mosaic virus (AMV), Cowpea mosaic virus (CPMV), Cucumber mosaic virus (CMV), Soybean dwarf virus (SbDV), Soybean mosaic virus (SMV), Soybean yellow common mosaic virus (SYCMV), Soybean yellow mottle virus (SYMMV), and Peanut stunt virus (PSV) (1). In 2012, Glycine max were observed in Daegu, South Korea, with mosaic and mottling symptoms on leaves. Samples with virus-like symptoms (n = 151) were collected from Daegu including legume genetic resource field. Virus particles were filamentous rod shaped, average length 760 nm, and were analyzed by RT-PCR using specific primers for several Potyviruses and previously reported viruses infecting soybean. Only two samples showing mosaic and mottling symptoms were identified as Clover yellow vein virus (ClYVV) based on RT-PCR using primers specific for ClYVV (5′-GTTGGCTTGGTTGACACTGA-3′ and 5′-CTTCGATCATGGATGCACA-3′). The sequences of amplified fragments were 97 to 98% similar with ClYVV. ClYVV is a distinct species in the genus Potyvirus and family Potyviridae. ClYVV is transmitted by several species of aphids and by mechanical inoculation (2). ClYVV was first reported on Gentiana scabra, and the disease has never been reported in soybean fields in Korea. The biological properties and full genome sequence of the selected ClYVV isolate of apparent virus symptoms between two samples were analyzed. The ClYVV isolate was inoculated to local lesion plants, re-isolated from local lesions three times, and propagated in Nicotiana benthamiana, and then named ClYVV-Gm. The ClYVV-Gm induced local lesions on inoculated leaves of N. tabacum cv. Xanthi-nc, Tetragonia expansa, and systemic symptoms on upper leaves of Chenopodium amaranticolor, C. quinoa, and N. clevelandii. The ClYVV-Gm caused mosaic and mottling symptoms on Glycine max cv. Kwangan and Phaseolus vulgaris. The genome of ClYVV-Gm was determined to be 9,584 nucleotides in length (GenBank Accession No. KF975894), and it shared 83% to 97% nucleotide identity with the sequences of 27 previously reported ClYVV isolates including Vicia fava and Pisum sativum. Despite low occurrence of ClYVV in Glycine max, ClYVV has a broad host range including tobacco, weed species, and soybean, which can lead to spreading of the virus. Our results indicate that emergence of ClYVV could become a problem to Leguminosae in Korea. To our knowledge, this is the first biological and molecular report of ClYVV infecting Glycine max in Korea. References: (1) Y. H. Lee et al. Korea Soybean Digest 29:7, 2012. (2) T. Sasaya et al. Phytopathology 87:1014, 1997.
pot and the roots washed with tap water. The number of tubercles was recorded on the root of each differential host. Race types were determined based on the reaction (tubercule formation on roots) of all the standard differential hosts to the test isolate. The results showed that races A, D, E,
Leonurus sibiricus L. (family Lamiaceae) has been used as a traditional herbal remedy to treat various gynecologic diseases. Although it is a widely distributed subtropical weed in Southeast Asia, L. sibiricus have been commercially cultivated on a small scale in many geographic areas of Korea. In August 2012, field-grown L. sibiricus plants showing mosaic, yellowing, and stunting symptoms were collected near a pepper field in Andong, Korea. Since L. sibiricus is only consumed as a raw material of traditional medicine in Korea, symptomatic plants lose commercial value entirely. To identify the causal agent(s) of the virus-like symptoms, total RNA was extracted from the symptomatic leaves, and a transcriptome library was generated using the TruSeq Stranded Total RNA with Ribo-Zero plant kit (Illumina, San Diego, CA) according to the standard protocol. Next-generation sequencing (NGS) was performed using an Illumina HiSeq2000 sequencer. De novo assembly of the quality filtered NGS reads (101-bp paired-end reads) were performed using the Trinity pipeline and the assembled contigs (92,329 contigs) were analyzed against the viral reference genome database in GenBank by BLASTn and BLASTx searches (3). The entire NGS procedure was performed by Macrogen Inc. (Seoul, South Korea). Among the analyzed contigs, only two large contigs were clearly of viral origin. Nucleotide blast searches showed that the first and second contigs (5,914 and 3,534 bp, respectively) have maximum identities of 91 and 95% to RNA1 of the isolate RP3 (GenBank Accession No. JX183225) and RNA2 of the isolate RP7 (JX183234) of Broad bean wilt virus 2 (BBWV-2), which were isolated from pepper in Korea. The NGS results were confirmed by analyzing the sequences of the fragments covering the entire BBWV-2 genome amplified by RT-PCR using specific primers for BBWV-2 as described previously (1). To obtain the complete genome sequence, terminal sequences of both RNA segments were analyzed by the 5′ and 3′ rapid amplification of cDNA ends (RACE) method as described previously (1). The assembled full-length sequences of BBWV-2 RNA1 and RNA2 isolated from L. sibiricus were 5,951 and 3,575 nucleotides in length, respectively, and deposited in GenBank under the accessions KM076648 and KM076649, respectively. BBWV-2 belongs to the genus Fabavirus in the family Secoviridae and it is known to have a wide host range. To investigate the host range of the BBWV-2 isolated from L. sibiricus, sap from the symptomatic leaves of L. sibiricus was inoculated to the test plants including Nicotiana benthamiana, Capsicum annuum (red pepper), and C. annuum var. gulosum (Paprika). RT-PCR detection and sequencing of the amplicons showed that all the inoculated test plants were infected with the BBWV-2 isolated from L. sibiricus. Currently, BBWV-2 is epidemic in pepper fields in Korea (1,2). Because BBWV-2 is easily transmitted by various aphids, and L. sibiricus is widely distributed in both wild and cultivated fields in Korea, this host might serve as a potential source of BBWV-2 to other crops such as pepper. To the best of our knowledge, this is the first report of BBWV-2 in L. sibiricus. References: (1) H.-R. Kwak et al. Plant Pathol. J. 29:274, 2013. (2) H.-R. Kwak et al. Plant Pathol. J. 29:397, 2013. (3) S.-E. Schelhorn et al. PLoS Comput. Biol. 9:e1003228, 2013.
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