Viroma de plantas de amendoim (Arachis hypogaea) provenientes do Estado de São Paulo

Abstract

The peanut (Arachis hypogaea L.) is an important source of protein and oil, and considered one of the most cultivated plants in Brazil and worldwide, that provide products consumed in different ways. It is the fourth world largest oleaginous in seed production, cultivated in over 30 countries, where China leads with 40.8%, followed by India with 14% and Nigeria with 7.5%. Brazil occupies the 17th position, concentrated in the Southeast and South regions, and the State of São Paulo accounts for 91.4% of national production. Among the factors that limit the development of this crop, there are several viruses, which cause reduction on productivity and the value of the product for marketing. Up to now, 31 viral species, classified in the following genera and respective families have been recorded naturally infecting peanut: Begomovirus (Geminiviridae), Bromovirus (Bromoviridae), Carlavirus (Betaflexiviridae), Cucumovirus (Bromoviridae), Irlavirus (Bromoviridae), Luteovirus (Luteoviridae), Pecluvirus (Virgaviridae), Potexvirus (Alphafleviridae), Potyvirus (Potyviridae), Rhabdovirus (Rhabdoviridae), Soymovirus (Caulimoviridae), Tospovirus (Bunyaviridae), Tymovirus (Tymoviridae) e Umbravirus (Tombusviridae). Of these genera, Potyvirus has more species. The advent of metagenomics, combined with high-performance technologies (Next Generation Sequencing - NGS) provides the exploration of micro-universe in various areas of science, including the Plant Virology. These techniques, together with bioinformatics, act as excellent tools for detection and characterization of novel viruses, as well as, all the viral genomes of different species or strains present in certain samples. The objective of this work was to study the virome of peanut plants exhibiting typical virus symptoms, obtained from producing areas of 10 counties of the State of São Paulo by NGS. The viral isolates were maintained by successive passages to peanut plants by grafting, under greenhouse conditions at the University of Brasilia (UnB). To perform the NGN, samples were semipurified aiming enrichment of virus particles, followed by total RNA extraction and sequencing carried out by Illumina platform. The metagenomic analysis permitted the detection the complete genome of Peanut mottle virus virus - PeMoV (Potyvirus) and Groundnut ringspot virus - GRSV (Tospovirus). Biological (indicator plants), serological (Dot-ELISA) and molecular (RT-PCR) tests were performed in order to characterize some GRSV isolates from peanut, which sequences were identified by metagenomic. Molecular analysis of the gene encoding the N protein, used for species taxonomy within Tospovirus genus, was also carried out with the selected isolates A1, N1 and O1. These three isolates induced symptomatic response in at least one of the indicator species (Datura stramonium L.) used in this work and widely cited in the literature. Positive results for these isolates were also confirmed by serology, indicating the presence of GRSV in peanut producing counties in the State of São Paulo. Phylogenetic analyzes of the segments L, M and S of tospoviruses revealed that the peanut GRSV isolates studied in this work grouped with sequences of GRSV isolate from watermelon (Citrullus lanatus (Thunb.) Matsum. & Nakai) and a recombinant isolate of GRSV and Tomato chlorotic spot virus - TCSV obtained from tomato (Solanum lycopersicum L.). However, the phylogenetic analysis using the N protein sequence, demonstrated that the present peanut isolate grouped with sequences of GRSV isolates previously studied in Brazil. After phylogenetic analysis for proteins of all GRSV segments, it is believed that the species GRSV and TCSV share the same M segment. The comparison of PeMoV coat protein sequences revealed that the Brazilian peanut isolate reported in this work is more phylogenetically related with the isolate from South Korea, obtained from soybean (Glycine max L.) with 98% identity. This is the first report in Brazil and in the world of the complete genome sequences of PeMoV and GRSV in peanuts, by using NGS.