Soybean (Glycine max L.) is a major food crop in Brazil. To investigate the occurrence of new viruses in soybean, ten leaf samples from the Pedra Petra/MT, Planaltina/DF and Luis Eduardo Magalhães/BA regions in Brazil were collected in January 2018 and characterized as a pooled sample by high throughput sequence analysis. Total RNA extraction was performed with the PureLink Viral RNA/DNA kit (Invitrogen) followed by library preparation and transcriptome sequencing using the Illumina HiSeq2500 platform. De novo assembly of the 27,674,874 reads was carried out using the CLC Genomics Workbench software v7.0.3. The 6,074 contigs obtained were submitted to a BLASTn search using the Geneious software v9.1.5. Results revealed the presence of three contigs showing high identities with groundnut ringspot orthotospovirus (GRSV) segments L (94% identity - KY350136), M (98% - KY350137) and S (97% - L12048). RT-PCR using orthotospovirus universal primers BR60/BR65 (Eiras et al. 2001) confirmed the presence of the virus in soybean with a fragment of the expect size (453 bp) obtained for GRSV isolate LEM from plants of cultivar M8349-IPRO showing crinkle/blistering leaves, mosaic and vein clearing in Luis Eduardo Magalhães/BA. Sanger sequencing of the amplicon showed a 99% nucleotide sequence identity with a GRSV isolate from peanut in Brazil (GenBank - KY400110). The complete nucleotide sequence of segments L, M and S was determined and consisted of 8,858 (MH686228), 4,847 (MH686227) and 3,040 (MH686229) bp, respectively. Further studies are necessary to evaluate the impacts of GRSV in soybeans in Brazil.
Reference
Eiras M, Resende RO, Missiaggia AA, Ávila ACD (2001) RT-PCR and dot blot hybridization methods for a universal detection of tospoviruses. Fitopatol Bras 26(2):170–175. https://doi.org/10.1590/S0100-41582001000200009
Acknowledgements
VHB, LFMW and FBS are recipients of CAPES and BRDM from CNPq PhD fellowships. RKS and MAP hold CNPq research fellowships. RKS is a grant recipient from FAPESP (Process Number 2017/ 21588-7).
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De Marchi, B.R., Bello, V.H., Watanabe, L.F.M. et al. Characterization and complete genome sequence of groundnut ringspot orthotospovirus in soybean in Brazil. J Plant Pathol 101, 401 (2019). https://doi.org/10.1007/s42161-018-0172-1
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DOI: https://doi.org/10.1007/s42161-018-0172-1