Development of molecular markers for western honey bee ( Apis mellifera L.) subspecies of regulatory concern in the United States

2026 · doi:10.64898/2026.03.05.709340
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⚙ AI-generated summary by claude@2026-07, 2026-07-15 ⓘ

This study developed and validated SNP-based real-time qPCR and RFLP assays targeting mitochondrial genes to accurately and cost-effectively identify western honey bee subspecies relevant to US regulations.

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⚙ AI-generated deep summary by claude@2026-07, 2026-07-15 · read from full text ⓘ

The study aimed to develop rapid, accurate, and cost-effective molecular markers to identify Apis mellifera subspecies of regulatory concern in the United States. Using established and novel real-time qPCR assays based on SNPs in mitochondrial cytochrome b (Cytb) and NADH dehydrogenase 4 (ND4) genes, the authors created a stepwise scheme to (1) distinguish bees of African (A-lineage) ancestry from other lineages, (2) detect African-derived honey bees, and (3) detect A. m. capensis in South Africa using an ND4 SNP, plus an ND2 RFLP assay to target a specific A-lineage clade. The paper reports validation of these markers for reliable subspecies assignation but does not present performance details beyond stating increased accuracy and that results are time- and cost-effective. The paper does not explicitly discuss endometriosis or adenomyosis; it was included in the corpus via a keyword match in the upstream search index.

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Abstract

Rapid, accurate and cost-effective identification of Apis mellifera subspecies is needed for subspecies of regulatory concern. We designed and validated subspecies markers based on single nucleotide polymorphisms (SNP) on mitochondrial cytochrome b ( Cytb ) and NADH dehydrogenase 4 ( ND4 ) genes. We used a combination of established and novel real-time qPCRs in a stepwise series, with increasing discrimination power, to (1) differentiate honey bees of African (A-lineage) ancestry from those of other lineages ( Cytb SNP #1), (2) identify African-derived honey bees (AHBs) ( Cytb SNP #2), and (3) detect A. m. capensis exclusively in the bee’s indigenous region of South Africa ( ND4 SNP). We also developed a restriction fragment length polymorphism assay targeting a SNP on the NADH dehydrogenase 2 ( ND2 RFLP) gene to detect the specific mitochondrial A-lineage clade. These assays allow for reliable time- and cost-effective results that provide increased accuracy on subspecies assignation.
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Abstract Rapid, accurate and cost-effective identification of Apis mellifera subspecies is needed for subspecies of regulatory concern. We designed and validated subspecies markers based on single nucleotide polymorphisms (SNP) on mitochondrial cytochrome b (Cytb) and NADH dehydrogenase 4 (ND4) genes. We used a combination of established and novel real-time qPCRs in a stepwise series, with increasing discrimination power, to (1) differentiate honey bees of African (A-lineage) ancestry from those of other lineages (Cytb SNP #1), (2) identify African-derived honey bees (AHBs) (Cytb SNP #2), and (3) detect A. m. capensis exclusively in the bee’s indigenous region of South Africa (ND4 SNP). We also developed a restriction fragment length polymorphism assay targeting a SNP on the NADH dehydrogenase 2 (ND2 RFLP) gene to detect the specific mitochondrial A-lineage clade. These assays allow for reliable time- and cost-effective results that provide increased accuracy on subspecies assignation. Competing Interest Statement The authors have declared no competing interest.

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europepmc
last seen: 2026-05-20T01:45:00.602351+00:00
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License: CC-BY-NC-ND-4.0