Page 60 - Read Online

Stuehler et al. Microbiome Res Rep. 2025;4:19 https://dx.doi.org/10.20517/mrr.2024.84 Page 13 of 16

In conclusion, The comprehensive screening and classification of Wolbachia in L. fagarae has broadened
the host spectrum of Wolbachia identified in members of Hemiptera: Psylloidea. A detailed MLST
classification of conserved housekeeping genes is standard practice and an effective method to determine
the phylogenetic classification of genetically divergent Wolbachia strains. The newly identified Wolbachia
strain in L. fagarae provides conserved biological elements that may be manipulated to influence host vector
fitness and fecundity. This study allows for the advancement of biological control applications that do not
involve the use of environmentally harmful chemicals, thus focusing on environmentally safer management
strategies for pest control while protecting the ecosystem.

DECLARATIONS
Acknowledgments
The authors thank Mrs. Maria Gonzalez, Biological Science Technician, ARS; Dr. Surya Saha,
Bioinformatics Advisor; and Dr. Thomson Paris, DOE/USDA, ORISE participant, sample collection. This
research was supported in part by an appointment to the Agricultural Research Service (ARS) Research
Participation Program administered by the Oak Ridge Institute for Science and Education (ORISE) through
an interagency agreement between the U.S. Department of Energy (DOE) and the U.S. Department of
Agriculture (USDA). ORISE is managed by ORAU under DOE contract number DE-SC0014664. All
opinions expressed in this paper are the authors’ and do not necessarily reflect the policies and views of
USDA, ARS, DOE, or ORAU/ORISE.

Authors’ contributions
Project design: Hunter WB
Conceptualization: Hunter WB, Stuehler DS Jr.
Methodology: Stuehler DS Jr., Cano LM, Qureshi JA
PCR screening and sequencing: Cano LM, Qureshi JA, Hunter WB
Molecular analysis: Stuehler DS Jr., Cano LM
Writing - original draft preparation: Stuehler DS Jr.
Writing - review and editing; Hunter WB, Cano LM
Figure preparation: Stuehler DS Jr., Hunter WB
All authors have read and agreed to the published version of the manuscript.

Availability of data and materials
The data supporting this work is provided in the text. Additional raw data will be made available by the
corresponding author upon reasonable request.

Financial support and sponsorship
The financial support for this study was provided to Hunter WB, Qureshi JA, and Cano LM by NIFA CAP
Grant 2020-70029-33176 “Therapeutic Molecule Evaluation and Field Delivery Pipeline for Solutions to
HLB”, NIFA USDA Citrus Greening award #2015-70016-23028, “Developing an Infrastructure and Product
Test Pipeline to Deliver Novel Therapies for Citrus Greening Disease”, and NIFA Hatch project FLA-IRC-
006515 “Deploying genome-informed pipelines and integrating transcriptome analyses of pathogen-host
interactions”. The Agricultural Research Service (ARS) Research Participation Program administered by the
Oak Ridge Institute for Science and Education (ORISE) through an interagency agreement between the US
Department of Energy (DOE) and the US Department of Agriculture (USDA), ORISE agreement, 60-6034-
1-002. ORISE is managed by ORAU under DOE contract number DE-SC0014664. All opinions expressed in
this study are the authors’ and do not necessarily reflect the policies and views of USDA, ARS, DOE, or
ORAU/ORISE.