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Outbreaks of emerging and re-emerging diseases in both animals and plants are increasing due to climate change, globalization, land-use change, and agricultural intensification. While most pathogen monitoring programs focus on zoonotic threats, wildlife and other organisms in natural habitats can also serve as reservoirs and sentinels for pathogens of agricultural and ecological concern. Plant communities and the pathogens circulating within them are underrepresented in integrated disease monitoring frameworks. This study demonstrates how biodiversity and zoonosis monitoring programs conducted in protected habitats (tallgrass prairies and woodlands) across Illinois, together with insect specimens preserved in biorepositories, can be leveraged to improve knowledge of the identities and ecological associations of a wide range of potential pathogens. We developed an integrative workflow combining taxonomic identification, molecular screening, and epidemiological inference to detect vector-borne plant pathogens from archived insect material. Focusing on Hemiptera (Auchenorrhyncha), we screened specimens for phytoplasmas (Mollicutes), uncultured bacterial plant pathogens transmitted by sap-feeding insects, and characterized host–pathogen associations. At least three distinct phytoplasma strains were detected: ‘Candidatus Phytoplasma asteris’ (16SrI-B), ‘Candidatus Phytoplasma pruni’-related strains (16SrIII), and ‘Candidatus Phytoplasma sacchari’-related strains (16SrXI-H). The latter represents the first documented occurrence of a 16SrXI-H phytoplasma subgroup in Illinois. Overall, we identified five insect specimens harboring phytoplasmas across four preserved sites, all of which were previously unreported for insect–phytoplasma associations. These findings demonstrate the value of existing biodiversity and zoonosis monitoring infrastructures for proactive surveillance of plant pathogens and extending the One Health paradigm to explicitly include natural ecosystems. 

Trivellone_etal_Full_PaperList_SystRev.xlsx: This dataset contains the list of peer-reviewed studies selected and critically appraised for a systematic review of quantitative PCR (qPCR) investigations tracking phytoplasma load dynamics in insect vectors. The dataset includes bibliographic information and selection status for each study, reflecting the inclusion and exclusion criteria applied during the review process. The literature search was completed on December 15, 2025. The list of inclusion and exclusion criteria are listed in the second spreadsheet. Further methodological details, including search strategy, screening workflow, and appraisal criteria, are described in the associated paper, “Tracking the early spatio-temporal dynamics of phytoplasma multiplication within its leafhopper vector”, as well as in the Supplementary Materials (see below), by Valeria Trivellone, Francesca Canuto, Giulia Lucetti, Christopher H. Dietrich, Luciana Galetto, Cristina Marzachì. 

Phytoplasmas are obligate bacterial pathogens transmitted by phloem-feeding insects and responsible for severe diseases in numerous crops worldwide. In Iran, insect-associated phytoplasma transmission pathways remain poorly resolved, particularly at fine phylogenetic and vector-specific scales. Here, we investigated phytoplasma strains detected in four plant species, grapevine (Vitis vinifera), soybean (Glycine max), barberry (Berberis vulgaris), and the weed Conyza canadensis, and in three potential insect vectors (Tropidocephala prasina, Eysarcoris ventralis, and Nysius graminicola) collected from distinct agroecosystems across Iran. Phytoplasmas were characterized by using nearly full-length 16S rRNA gene sequences and a multilocus dataset of protein-coding genes obtained through a targeted next-generation sequencing approach. Five phytoplasma strains belonging to ribosomal groups 16SrI, 16SrVI, 16SrIX, and 16SrXII were identified, including two novel ribosomal subgroups, 16SrI-AS and 16SrIX-K. Several previously unreported plant–phytoplasma and insect–phytoplasma associations were documented. Comparative phylogenetic analyses revealed that ribosomal and multilocus markers capture complementary evolutionary signals, with protein-coding genes providing additional resolution beyond 16S-based classification. These findings highlight the potential role of diverse hosts and polyphagous insects, not yet confirmed as vectors, in phytoplasma circulation and underscore how high-throughput next-generation sequencing and multilocus approaches advance our understanding of phytoplasma diversity and evolution. 

Phytoplasmas are a group of plant-pathogenic, cell-wall-less bacteria vectored primarily by leafhoppers (Hemiptera Cicadellidae), one of the most diverse families of insects. Despite the importance of documenting associations between phytoplasmas, their insect vectors, and plant hosts to prevent disease outbreaks, such knowledge is currently highly incomplete and largely neglects the diversity of the system in natural areas. Here, we used anchored hybrid enrichment (AHE) to recover the DNA of five plant genes (rbcL, matK, ITS1, ITS2, and trnH-psbA) in 58 phloem-feeding leafhoppers from around the world that had previously tested positive for phytoplasma infection. Using BLASTn and a strict filtering approach, we assigned taxonomic classifications to the plant sequences and tested for cophylogenetic signals between potential Deltocephalinae leafhopper vectors and their associated plants. We observed incongruence between plant and insect phylogenies. Many leafhopper species, including presumed grass specialists, fed on distantly related plant lineages; 66% of sampled leafhoppers fed on plants from at least two different orders. By disentangling phytoplasma–leafhopper–plant interactions, we identify locations at risk of phytoplasma disease outbreaks. Furthermore, the observed wide diet breadth raises questions about how phytoplasma infection may manipulate the feeding preference of their insect host and helps fill the gaps in understanding the ecology and diversification of the tripartite association. 

Based on study of the holotype of Scaphytopius (Tumeus) rubidus DeLong, 1980, it was determined that this species does not belong in any genus of the subfamily Deltocephalinae described so far. To accommodate this species, we describe and illustrate a new genus in the tribe Bahitini, Anavilhanas gen. nov., and propose the new combination, Anavilhanas rubida (DeLong) comb. nov.