Emerging disease in melon crop associated with a mixed infection by three whitefly-transmitted viruses in La Comarca Lagunera, Mexico

Abstract

Background/Objective. Cucurbits are an economic pillar of global and national agriculture. However, their productivity is threatened by the increasing incidence of viral plant diseases. Consequently, coinfections with members of the Begomovirus and Crinivirus genera have been associated with exacerbated symptoms and high incidence rates in crops, leading to emerging disease patterns. The aim of this study was to identify and molecularly characterize the begomoviruses and criniviruses associated with an emerging disease in melon crops in a municipality of the Comarca Lagunera region, Mexico. Materials and Methods. A targeted sampling of nine melon plants exhibiting symptoms such as severe interveinal chlorosis, mottling, yellow mosaic patterns, and fruit deformation was carried out. For viral detection, DNA extraction was performed using 3% CTAB and RNA extraction using the TRIzol reagent. Begomoviruses and criniviruses were detected using PCR and RT-PCR with specific primers. Complete begomovirus genomes were enriched by rolling circle amplification (RCA). Amplicons derived from Open Reading Frame (ORF) of the capsid protein (CP) of criniviruses were generated using RT-PCR. Subsequently, the genomes and CP amplicons were cloned and sequenced for phylogenetic and evolutionary analysis. Results. Molecular identification revealed the prevalence of mixed infections in seven of nine samples, comprised of two begomovirus species, cucurbit leaf crumple virus (CuLCrV, Begomovirus cucurbitae) and watermelon chlorotic stunt virus (WmCSV, Begomovirus citrulli), and the crinivirus cucurbit yellow stunting disorder virus (CYSDV, Crinivirus cucurbitae). Genomic analyses confirmed that CuLCrV and WmCSV possess genomic structure typical of begomoviruses. Significant divergence was observed at specific loci (CuLCrV C4 gene, WmCSV NSP gene, and DNA-B intergenic regions of both species), suggesting their potential involvement in evolutionary adaptation processes. Phylogenetically, although the CuLCrV isolate showed close relationship to isolates from Arizona and Sonora, it was placed in a clade distinct from those previously reported in the region, indicating an independent evolutionary course. Furthermore, phylogenetic analysis based on the nucleotide sequence of the CYSDV capsid protein (CP) gene revealed a close relationship with isolates previously reported in Jordan and Mexico. Conclusions. This study represents the first report in Mexico of a disease associated with WmCSV, CuLCrV, and CYSDV coinfection in cucurbits. The findings amphasize the complexity of emerging diseases and highlight the importance of strengthening molecular epidemiological surveillance to mitigate the impact of these mixed infections on Mexican agriculture.