Detecting bumble bees in the wild using environmental DNA: Development and validation of a qPCR assay for the endangered Franklin’s bumble bee (Bombus franklini)

Genome
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Abstract

Environmental DNA (eDNA) sampling is a noninvasive alternative to conventional methods of surveying insects that may be particularly useful for detecting pollinators. We developed a quantitative polymerase chain reaction (qPCR) assay to detect the DNA of Franklin’s bumble bee (Bombus franklini) from flower samples and conducted an initial test of the assay using samples collected within and around the historical range of the species. We further analyzed all samples using metabarcoding. Our qPCR assay successfully amplified B. franklini DNA and exhibited no cross-reactivity with nontarget bumble bee DNA during in silico and in vitro testing. We did not detect B. franklini DNA from field-collected flower samples using either qPCR or metabarcoding. However, metabarcoding analysis revealed DNA of at least 16 other bumble bee species. This finding underscores the potential utility of eDNA sampling for surveying bumble bees. Nondetection of B. franklini from field-collected flower samples may be due to the extreme rarity of the species; B. franklini is endangered and has not been observed in the wild since 2006. Our B. franklini assay is among the first bee-specific qPCR assays ever developed and provides proof of concept for additional assays that may improve detection rates of rare and endangered bees.

Suggested Citation

Grossklaus, M.R., Pilliod, D.S., Spear, S.F., Laramie, M.B., Kholwadwala, A., Boone, A., Lor, Y., Kaminski, M., and Everett, J.G., 2026, Detecting bumble bees in the wild using environmental DNA: Development and validation of a qPCR assay for the endangered Franklin’s bumble bee (Bombus franklini): Genome, v. 69, 13 p., https://doi.org/10.1139/gen-2025-0006.

Study Area

Publication type Article
Publication Subtype Journal Article
Title Detecting bumble bees in the wild using environmental DNA: Development and validation of a qPCR assay for the endangered Franklin’s bumble bee (Bombus franklini)
Series title Genome
DOI 10.1139/gen-2025-0006
Edition Online First
Volume 69
Year Published 2026
Language English
Publisher Canadian Science Publishing
Contributing office(s) Forest and Rangeland Ecosystem Science Center
Description 13 p.
Country United States
State California, Oregon
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