Author ORCID Identifier

https://orcid.org/0009-0007-6054-9133

Date Available

7-21-2023

Year of Publication

2023

Degree Name

Master of Science (MS)

Document Type

Master's Thesis

College

Agriculture, Food and Environment

Department/School/Program

Entomology

First Advisor

Dr. Lynne Rieske-Kinney

Abstract

Laurel wilt disease (LWD) is a lethal vascular disease impacting lauraceous hosts caused by Harringtonia lauricola, the fungal symbiont of the redbay ambrosia beetle (Xyleborus glabratus Eichoff) (RAB) (Coleoptera: Curculionidae). LWD has caused tree mortality throughout the southeastern United States and is continuing to spread into new regions. Current management methods have not been successful in preventing spread, warranting investigation into innovative techniques including RNA interference (RNAi).

Elongation factor-1 alpha (ef1a) and actin (act) were established as stably expressed reference genes after exposing beetles to different photoperiod, temperature, and dsRNA exposure. After RAB oral ingestion of double- stranded RNAs (dsRNAs) of three target genes (hsp, shi, and iap), mortality and changes in gene expression were assessed. Following exposure, significant mortality occurred in all three genes. Quantitative PCR (qPCR) was used to investigate changes in gene expression at different timepoints with significant changes occurring in all three target genes.

Additionally, working in sassafras forests at the leading edge of the LWD range expansion, insects were evaluated for internal and phoretic presence of H. lauricola. Results show internal presence of H. lauricola in one hidden snout weevil and two granulate ambrosia beetles, demonstrating that other insects can potentially vector LWD.

Digital Object Identifier (DOI)

https://doi.org/10.13023/etd.2023.294

Funding Information

This research was supported by funds provided by the United States Department of Agriculture Animal and Plant Health Inspection Service AP20PPQS & T00C061, the University of Kentucky, and the Kentucky Agricultural Experiment Station under McIntire-Stennis 2352657000 in 2021.

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