First report of milk vetch dwarf virus infecting faba bean in Jiangsu province in China
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First report of milk vetch dwarf virus infecting faba bean in Jiangsu province in China Kun Zhang 1 & Hongmei Xu 1 & Ying Zang 1 & Jiahuan Chen 2 & Xinjian Zhuang 1 & Zhen He 1 Received: 12 June 2019 / Accepted: 29 January 2020 # Società Italiana di Patologia Vegetale (S.I.Pa.V.) 2020
Keywords Milk vetch dwarf virus . Faba bean . Nanovirus . China
Milk vetch dwarf virus belongs to the genus Nanovirus. Milk vetch dwarf virus (MDV) causes dwarf, chlorotic, and crinkle symptoms, and was identified in several plant species in China (Kamran et al. 2019; Zhang et al. 2017). The genome of MDV contains eight different ssDNA components, which are denoted DNA-R, -S, −M, -C, -N, -U1, -U2, and -U4 (Kamran et al. 2019; Sano et al. 1998). MDV mainly infects legumes, and is transmitted by several aphid species colonizing legumes (Vetten 2008). In 2018, faba bean plants, which showed mosaic, yellowing, stunting, and crinkle, were collected from city of Yangzhou and its suburbs in Jiangsu Province. The total RNA was isolated from three plants by Trizol Reagent (Invitrogen, CA, USA), and the siRNAs were recycled after separation by urea-denaturing PAGE. The NEBNext Ultra II RNA Library Prep Kit for Illumina (NEB, UK) was used for siRNA library construction. The library was sequenced by pair-end method at Illumina Hiseq 2000 platform (Sangon, Shanghai, China). The raw data were processed with Trinity software, and clean reads (21,024,193 total reads) were de novo assembled by Velvet (Zerbino 2010). Among the 131 assembled contigs, 53 contigs matched to eight different components of MDV. Based on the sequences of the matched contigs, primer pairs were designed and used for the
amplification of the MDV components (Supplementary Table SI). The genomic DNA was extracted from the positive samples and was taken as template of PCR. The eight DNA fragments were cloned into pMD19-T vector (Takara, Dalian, Japan) and three independent clones of each component were sequenced. The obtained consensus sequences (GenBank accession Nos. MN053907, MN053908, MN437517, MN437518, MN437519, MN437520, MN437521, and MN437522) have 98.7%, 97.2%, 91.5%, 91.4%, 95.2%, 97.1%, 90.3%, and 88.8% similarity to MDV Anhui isolate DNA-R (KY070246), -S (KY070245), -C (KY070240), −M (KY070244), -N (KY070242), -U1 (KY070241), -U2 (KY070243), and –U4 (KY070247), respectively (Supplementary Table SII). On the basis of our siRNA deep sequencing and PCR validation with MDV specific primers, the chlorosis, mosaic, stunning, and dwarf phenotype of the collected faba bean plants was most likely caused by MDV infection. Funding information This work was funded by the National Natural Science Foundation of China (31801699), Natural Science Foundation of the Jiangsu Province (BK20180904), Natural Science Foundation of the Jiangsu Higher Education Institutions (18KJB210012), and Youth Science and Technology Talents Lifting Project of Jiangsu Province.
Electronic supplementary material The online version of this article (https://doi.org/10.1007/s42161-020-00511-8)
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