Publications

4682

In 2024, an infestation of corn leafhoppers, Dalbulus maidis (DeLong) (Hemiptera: Cicadellidae), was detected in several commercial corn fields across Oklahoma (OK) (Faris et al. 2024). Corn hybrids in the infested field showed symptoms resembling maize bushy stunt, including chlorotic striping, reddening of leaves, production of multiple ears, and stunting. Visual disease incidence ranged from 25% to 100% among affected fields. Leaf samples from symptomatic plants were collected from 15 commercial corn fields across seven counties in OK (Payne, Logan, Caddo, Kay, Texas, Pottawatomie, and Grady) during September, October, and November of 2024. Each sample consisted of 3-4 symptomatic corn leaves randomly collected in each field. In addition, 43 corn leafhopper samples were collected from the symptomatic fields using handheld vacuums. Each corn leafhopper sample consisted of a minimum of 10 corn leafhoppers per field. In the laboratory, the corn leafhoppers were identified as D. maidis based on morphological characteristics as described (Faris et al. 2024). DNA from the 15 leaf samples was extracted from the leaf midrib and blade tissues using the DNeasy® Plant Mini Kit. Insect DNA was extracted using a CTAB-based method (Gorayeb et al. 2023), with each sample consisting of five pooled insects per field. DNA samples from plants and insects were initially screened using the R16F2n/R16R2 primer pair (Gundersen and Lee, 1996) for the presence of phytoplasmas. Seventeen insect and five plant samples tested positive for phytoplasma. Subsequent phytoplasma screening was performed on insect and plant samples collected from the same fields on the same dates, comprising a total of four insect and five plant samples. To further characterize the phytoplasma, a semi-nested PCR targeting the 16S rRNA gene was performed on these samples (four insect and five plant samples) using primer pairs P1/16S-SR followed by P1A/16S-SR (Wei et al. 2021), yielding an expected amplicon of approximately 1.5 Kb. An additional gene marker, the ribosomal protein (rp) gene, was also amplified using universal primers rpF1/rpR1 (Lee et al. 1998), producing 1.2 Kb PCR amplicons. PCR products were subjected to Sanger sequencing, and 16S rRNA and rp gene sequences were deposited in GenBank under accession numbers PX927497–PX927505 and PX993703-PX993711, respectively. BLASTn analysis of the 16S rRNA sequences showed 99.14–99.93% nucleotide identity with reference strains of ‘Candidatus Phytoplasma asteris’ associated with maize bushy stunt (accessions MW578284 and CP015149). Similarly, the rp gene sequences shared 99.91–100% nucleotide identity with the same reference strains. Using the iPhyClassifier tool (Zhao et al. 2013), the 16S rRNA gene sequences were classified within 16Sr group I, subgroup B, with a similarity coefficient of 1.0 (AP006628). The pairwise comparison and subsequent phylogenetic analysis showed that the OK isolates clustered within the 16SrI-B clade. This finding represents the first report of a 16SrI-B phytoplasma (‘Candidatus Phytoplasma asteris’) associated with maize bushy stunt in OK. Although currently classified as ‘Candidatus Phytoplasma asteris’, a proposal to establish a novel phytoplasma species, ‘Candidatus Phytoplasma zeae’, has been made based on ecological distinctiveness, vector specificity, whole-genome comparisons, and community consensus (Pellegrinetti et al. 2025).

‘Candidatus Liberibacter asiaticus’ (CLas), the causal agent of citrus huanglongbing, is transmitted by the Asian citrus psyllid Diaphorina citri. While CLas-positive (CLas+) females exhibit increased fecundity and metabolic demands, their neuroendocrine regulation mechanisms remain unclear. We propose CLas manipulates dopamine (DA) signaling to enhance psyllid fecundity and CLas proliferation. Metabolomics revealed elevated DA in CLas+ females. Silencing DA synthesis genes and receptor DcDop2 via RNAi reduced lipid reserves, fecundity, and ovarian CLas titers. Through combined in vivo and in vitro experiments, we demonstrated that the microRNA miR-31a suppresses DcDop2 expression by binding to its 3’ untranslated region. Overexpression of miR-31a resulted in decreased DcDop2 expression and CLas titers in the ovaries, eliciting phenotypic defects akin to DcDop2 knockdown. Furthermore, DcDop2 knockdown and miR-31a overexpression reduced juvenile hormone (JH) levels and adipokinetic hormone (AKH) signaling in fat bodies and ovaries. Consequently, CLas regulates the DA-DcDop2 signaling axis to improve D. citri lipid metabolism and fecundity, while simultaneously promoting its replication. These findings reveal a coevolution between CLas proliferation and ovarian development in the insect host. This discovery enhances our understanding of the molecular interplay between plant pathogens and vector insects and offers novel targets and strategies for HLB field management.

Abstract De novo assembly of ancient and modern bacterial metagenomes can shed light on evolution and ecology of bacterial species that are challenging to culture. Tannerella and Porphyromonas are bacterial genera linked to periodontal disease, and understanding their evolution may reveal insights into their role in oral disease development. We performed pangenomic and phylogenetic analyses on a global set of isolates and metagenome-assembled genomes of the genera Tannerella (n = 238) and Porphyromonas (n = 976), including 66 genomes from ancient dental calculus samples (up to 14,800 years old), and modern oral samples from present-day living populations. We identify a novel species of oral Tannerella in modern and ancient humans, which we call Ca. Tannerella abscondita, that is related to and often mistaken for Tannerella forsythia but differs in its virulence repertoire. We reveal distinct niche tropism in Tannerella species and Porphyromonas pasteri, but not Porphyromonas gingivalis. There is limited phylogeographic structuring, and virulence genes are homogeneously distributed across continents and oral niches. Saliva-derived strains of T. forsythia and P. gingivalis from Oceania and T. serpentiformis and P. pasteri from Asia show enrichment of pseudogenes related to ecological niche transitions. A phylogenetic analysis of the P. gingivalis major fimbrial protein gene fimA reveals the genes cluster by genotypes, and that no ancient genes are found in genotypes I and Ib. Using de novo assembly for bacterial pangenomics improves the representation of oral genera found in reference databases and enhances our ability to study the evolutionary history of these taxa.

Abstract Since 2025, EFSA has conducted expert knowledge elicitations to estimate survey parameters for the detection of EU priority pests listed in Commission Delegated Regulation (EU) 2019/1702, in cases where pest‐specific information is lacking in the scientific literature. The design and implementation of statistically sound and risk‐based surveys require quantitative parameters such as sampling effectiveness, design prevalence and relative risk. This report presents the estimated values for Bactericera cockerelli and ‘Candidatus Liberibacter solanacearum’, together with the supporting evidence and expert rationale. These values will be integrated into the EFSA Plant Pest Survey Toolkit to guide EU Member States in designing harmonised pest surveys across the EU.