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ARS Home » Northeast Area » Beltsville, Maryland (BARC) » Beltsville Agricultural Research Center » Bee Research Laboratory » Research » Publications at this Location » Publication #372514

Research Project: Managing Honey Bees against Disease and Colony Stress

Location: Bee Research Laboratory

Title: Non-invasive genotyping of honey bee queens Apis mellifera L.: Transition of the DraI mtDNA COI-COII test to in silico

item Madella, Shayne
item Grubbs, Kyle
item Alburaki, Mohamed

Submitted to: Insects
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 12/27/2020
Publication Date: 12/30/2020
Citation: Madella, S., Grubbs, K.F., Alburaki, M. 2020. Non-invasive genotyping of honey bee queens Apis mellifera L.: Transition of the DraI mtDNA COI-COII test to in silico. Insects. 12(1):19.

Interpretive Summary: The honey bee Apis mellifera L. colony is headed by a single and indispensable queen whose duty is to ensure brood production and provide pheromonal stability within the colony. Thus, identification of the queen’s genetic background is usually conducted on samples of her offspring at various developmental stages. In this study, we present a technical method which allows the identification of the queen maternal lineage and subspecies using the remainder tissue of her clipped wing. The DraI mtDNA COI-COII (DmCC) test was applied to various sizes of queen and worker wings and the results were compared with results obtained from other bee tissues such as legs, head and thorax. Furthermore, we proposed a new method allowing in silico transition of the DmCC test and haplotype identification based on extended sequencing of the tRNAleu and COII. Our results show that DNA extracted by Chelex 10% from one-third of a queen wing is deemed adequate for a successful identification of her maternal evolutionary lineage, haplotype and subspecies. The in silico method proposed in this study fully adheres to the established guidelines of the DmCC and provides a universal standard for haplotype identification. Moreover, this method offers faster and more precise results by reconciling both Cleaved Amplified Polymorphisms CAPS and sequencing approaches.

Technical Abstract: In this study, we tested a quick and straightforward method to extract DNA from the remainder of queen clipped wings. The adequacy of the extracted DNA for genetic studies and sequencing was tested. Finally, we detailed a new method that offers a full in silico transition of the DmCC test, cutting significant lab work while preserving the DmCC test’s rules for haplotype identification.