1 First report of Lettuce chlorosis virus infecting periwinkle in Brazil 2 3 Gabriel M. Favara¹, Viviana M. Camelo-García¹, David M. A. Spadotti¹, João M. F. 4 Silva², Tatsuya Nagata², Elliot W. Kitajima¹, Jorge A. M. Rezende¹ 5 6 ¹Departamento de Fitopatologia e Nematologia, Escola Superior de Agricultura Luiz de 7 Queiroz, Universidade de São Paulo, Piracicaba, SP, Brazil; ²Departamento de Biologia 8 Celular, Universidade de Brasília, Brasília, DF, Brazil 9 10 The occurrence of periwinkle (Catharanthus roseus) plants exhibiting symptoms 11 associated with virus infection is very common in Brazil (Maciel et al. 2011). In 2018, a 12 plant of C. roseus with symptoms of mosaic and leaf malformation from a residential 13 garden in Piracicaba city, state of São Paulo, Brazil was analyzed by high-throughput 14 sequencing (HTS). Total RNA was extracted from symptomatic leaves of C. roseus using 15 the Purelink® viral RNA/DNA kit (Thermo Fisher Scientific, Waltham, USA). A cDNA 16 library was prepared using the TruSeq Stranded Total RNA Sample Prep Kit (Illumina, 17 San Diego, USA). The cDNA was sequenced by the Illumina HiSeq 2500 System at the 18 Center of Functional Genomics (ESALQ/USP, Piracicaba, Brazil), using a HiSeq Flow 19 Cell v4 and the HiSeq SBS Kit v4 in paired-end mode, producing reads of 100 bases. A 20 total of 58,135,090 reads were generated. Trimming and removal of adapter sequences 21 was performed with BBDuk (Bushnell) and de novo assembly was performed with 22 MEGAHIT v1.1.3 (Li et al. 2016). Viral contigs were identified through tBLASTx 23 searches (Altschul et al. 1990) against the Virus RefSeq database from NCBI, which led 24 to the identification of a potyvirus, Catharanthus mosaic virus (CatMV), previously 25 reported to infect periwinkle in Brazil (Maciel et al. 2011), and the crinivirus Lettuce 26 chlorosis virus (LCV), characterized by Duffus et al. (1996) and previously identified 27 from periwinkle in China (Tian et al. 2017). The near-complete genome of RNA-1 (8,604 Page 1 of 4