Agricultural pests can cause direct damage to crops, including chlorosis, loss of vigor, defoliation, and wilting. In addition, they can also indirectly damage plants, such as by transmitting pathogenic micro-organisms while feeding on plant tissues, affecting the productivity and quality of crops and interfering with agricultural production. Among the known arthropod pests, mites are highly prevalent in global agriculture, particularly those from the Tetranychidae family. The two-spotted spider mite, Tetranychus urticae, is especially notorious, infesting about 1600 plant species and causing significant agricultural losses. Despite its impact on agriculture, the virome of T. urticae is poorly characterized in the literature. This lack of knowledge is concerning, as these mites could potentially transmit plant-infecting viral pathogens, compromising food security and complicating integrated pest management efforts. Our study aimed to characterize the virome of the mite T. urticae by taking advantage of publicly available RNA deep sequencing libraries. A total of 30 libraries were selected, covering a wide range of geographic and sampling conditions. The library selection step included selecting 1 control library from each project in the NCBI SRA database (16 in total), in addition to the 14 unique libraries from a project containing field-collected mites. The analysis was conducted using an integrated de novo virus discovery bioinformatics pipeline developed by our group. This approach revealed 20 viral sequences, including 11 related to new viruses. Through phylogenetic analysis, eight of these were classified into the Nodaviridae, Kitaviridae, Phenuiviridae, Rhabdoviridae, Birnaviridae, and Qinviridae viral families, while three were characterized only at the order level within Picornavirales and Reovirales. The remaining nine viral sequences showed high similarity at the nucleotide level with known viral species, likely representing new strains of previously characterized viruses. Notably, these include the known Bean common mosaic virus (BCMV) and Phaseolus vulgaris alphaendornavirus 1, both of which have significant impacts on bean agriculture. Altogether, our results expand the virome associated with the ubiquitous mite pest T. urticae and highlight its potential role as a transmitter of important plant pathogens. Our data emphasize the importance of continuous virus surveillance for help in the preparedness of future emerging threats.

Theobroma cacao plantations are of significant economic importance worldwide, primarily for chocolate production. During the harvest and processing of cocoa beans, they are subjected to fermentation either by microorganisms present in the environment (spontaneous fermentation) or the addition of starter cultures, with different strains directly contributing distinct flavor and color characteristics to the beans. In addition to fungi and bacteria, viruses are ubiquitous and can affect the quality of the fermentation process by infecting fermenting organisms, destabilizing microbial diversity, and consequently affecting fermentation quality. Therefore, in this study, we explored publicly available metatranscriptomic libraries of cocoa bean fermentation in Limon Province, Costa Rica, looking for viruses associated with fermenting microorganisms. Libraries were derived from the same sample at different time points: 7, 20, and 68 h of fermentation, corresponding to yeast- and lactic acid bacteria-driven phases. Using a comprehensive pipeline, we identified 68 viral sequences that could be assigned to 62 new viral species and 6 known viruses distributed among at least nine families, with particular abundance of elements from the Lenarviricota phylum. Interestingly, 44 of these sequences were specifically associated with ssRNA phages (Fiersviridae) and mostly fungi-infecting viral families (Botourmiaviridae, Narnaviridae, and Mitoviridae). Of note, viruses from those families show a complex evolutionary relationship, transitioning from infecting bacteria to infecting fungi. We also identified 10 and 3 viruses classified within the Totiviridae and Nodaviridae families, respectively. The quantification of the virus-derived RNAs shows a general pattern of decline, similar to the dynamic profile of some microorganism genera during the fermentation process. Unexpectedly, we identified narnavirus-related elements that showed similarity to segmented viral species. By exploring the molecular characteristics of these viral sequences and applying Hidden Markov Models, we were capable of associating these additional segments with a specific taxon. In summary, our study elucidates the complex virome associated with the microbial consortia engaged in cocoa bean fermentation that could contribute to organism/strain selection, altering metabolite production and, consequently, affecting the sensory characteristics of cocoa beans.

The majority of giant algal viruses belong to the family Phycodnaviridae, class Algavirales, phylum Nucleocytoviricota. Among them, the genus Chlorovirus is the most studied, with three recognized groups based on genomics and host range, although many fundamental questions remain to be elucidated, particularly regarding their diversity. In this study, we focus on betachloroviruses, a poorly explored subgroup that infects the alga Micractinium conductrix Pbi. Here, we describe the isolation and genomic analysis of 11 new betachloroviruses from water samples collected in Nebraska, USA. With 25 fully sequenced genomes now available, we assessed the genomic diversity of these viruses. They have double-stranded DNA genomes ranging from 295 to 374 kbp, encoding hundreds of ORFs, of which a large number (~40%) lack known function. Comparative genomics and phylogenetic analyses revealed three species of betachlorovirus, each with high intra-species genomic identity. Notably, some isolates with over 99.5% genomic identity display markedly different plaque phenotypes, which led us to propose the use of the term genomovar among giant algal viruses, a concept potentially applicable to other giant viral groups yet to be explored. Altogether, this work advances our understanding of betachloroviruses and highlights the importance of linking viral genotype to phenotype, opening new avenues for exploring the diversity of giant algal viruses.

Cocoa (Theobroma cacao L.) is a major agricultural commodity, essential for the global chocolate industry and the livelihoods of millions of farmers. However, viral diseases pose a significant threat to cocoa production, with Badnavirus species causing severe losses in Africa. Despite its economic importance, the overall virome of T. cacao remains poorly characterized, limiting our understanding of viral diversity and potential disease interactions. This study aims to assess the cocoa-associated virome by analyzing 109 publicly available RNA-seq libraries from nine BioProjects, covering diverse conditions and geographic regions. We implemented a comprehensive bioinformatics pipeline integrating multiple viral sequence enrichment steps, a hybrid assembly strategy using different assemblers, and sequence similarity searches against NCBI non-redundant databases. Our approach identified ten putative novel viruses associated with the cocoa microbiome and a novel Badnavirus species. These findings provide new insights into the viral landscape of T. cacao, characterizing the diversity of cacao-associated viruses and their potential ecological roles. Expanding the catalog of viruses associated with cocoa plants not only enhances our understanding of plant–virus–microbiome interactions but also contributes to the development of more effective disease surveillance and management strategies, ultimately supporting sustainable cocoa production.

Endogenous Viral Elements (EVEs) are viral sequences integrated into the host germline and passed to offspring. Most virus types can integrate, often with

MicroRNAs (miRNAs) are key post-transcriptional regulators involved in a wide range of biological processes in insects, yet little is known about their roles in stingless bees. Here, we present the first characterization of miRNAs in Melipona quadrifasciata using small RNAs (sRNAs) deep sequencing. A total of 193 high-confidence mature miRNAs were identified, including 106 M. quadrifasciata-exclusive sequences. Expression profiling revealed that mqu-miR-1 and mqu-miR-276 together accounted for over 70% of all miRNA reads, suggesting their central roles in development and reproduction. Comparative analyses showed a higher conservation of M. quadrifasciata miRNAs with other Hymenopterans, especially Apis mellifera and Bombus spp. Putative target genes were predicted using a consensus approach, and functional annotation indicated their involvement in diverse biological regulatory pathways. This work represents the first comprehensive identification of the miRNA repertoire in stingless bees using sRNAs and provides a valuable foundation for understanding miRNA-mediated gene regulation in this ecologically and economically important pollinator.