Suchergebnisse

We assembled the mitogenome of Apis mellifera siciliana, which was previously identified as African by the tRNA-leu-cox2 intergenic region. The mitogenome is 16,590 bp long. The gene content and organization are identical to other A. mellifera mitogenomes, containing 13 protein-coding genes, 22 transfer RNA genes, and 2 ribosomal RNA genes. Phylogenetic analysis showed a close mitochondrial relationship between A. m. siciliana and other African subspecies such as Apis mellifera sahariensis, Apis mellifera intermissa, and Apis mellifera ruttneri. ; This work was supported by MEDIBEES - Monitoring the Mediterranean Honey Bee Subspecies and their Resilience to Climate Change for the Improvement of Sustainable Agro-Ecosystems; BEEHAPPY ([POCI-01-0145- FEDER-029871]; FCT and COMPETE/QREN/EU). MEDIBEES is part of the PRIMA program supported by the European Union. Fundac¸~ao para a Ciência e a Tecnologia (FCT) provided financial support by national funds (FCT/MCTES) to CIMO [UIDB/00690/2020]. Dora Henriques is supported by the project BEEHAPPY ([POCI-01-0145-FEDER-029871]; FCT and COMPETE/QREN/EU). ; info:eu-repo/semantics/publishedVersion

This work has received funding from the Programa Nacional Apícola 2020-2022 under the project "AUTENT+ Desenvolvimento de abordagens inovadoras com vista à valorização e exploração do potencial de mercado do mel Português". The authors are also grateful to the Foundation for Science and Technology (FCT, Portugal) for financial support by national funds FCT/MCTES to CIMO (UIDB/00690/2020). D. Henriques is supported by the project BeeHappy (POCI-01-0145-FEDER-029871) funded by FEDER (Fundo Europeu de Desenvolvimento Regional) through the program COMPETE 2020—POCI (Programa Operacional para a Competividade e Internacionalização), and by Portuguese funds through FCT and A.R. Lopes by the PhD scholarship funded by the FCT (SFRH/BD/143627/2019). ; Honey is a natural product widely consumed around the globe, not only for its taste and nutritional value, but also for its health benefits. Being a product of high dietary relevance and increasing demand, it has also become a target of economically motivated adulteration. According to the 2014 European Parliament report on the food crisis, fraud in the food chain and the control thereof, honey is among the 10 food products most prone of being adulterated [1]. Up until now, honey authenticity was mainly focused on the issues of sugars addition and botanical and geographical origin. However, recently an increased attention has been paid to the entomological origin of honey. To this aim, different approaches have been proposed to differentiate honey produced by different Apis mellifera subspecies, including those from distinct mitochondrial (mt) DNA lineages [2]. This work aimed to develop a novel real-time PCR method coupled with HRM analysis that allows for the simultaneous differentiation of honeybee from maternal lineages A, M and C, for further application in honey authentication. In this sense, data previously obtained from the mitogenomes of a total of 112 honeybees of different lineages were considered for the development of new DNA markers. Considering the aim of further application in honey, new primer sets were designed to amplify short fragments that included different single nucleotide polymorphisms (SNPs) allowing for HRM application. Three primer sets were proposed, amsCOI-F/amsCOI-R targeting the Cytochrome oxidase I (COI) gene, amsND1-F-amsND1-R targeting the NADH-ubiquinone oxidoreductase chain 1 (ND1) gene and amsCox3-F/amsCox3-R targeting the Cytochrome oxidase subunit III (Cox3) gene. Each primer set was first tested using qualitative PCR using DNA extracted from honeybees of A, M and C mtDNA lineages. After optimizing the real-time PCR conditions, each primer set was tested using a series of mtDNA extracted from honeybees. While amsCOI-F/amsCOI-R allowed only for the separation of the honeybees in two clusters, with lineage C and M clustering together, both the amsCox3-F/amsCox3-R and amsND1-F/amsND1-R set of primers allowed to differentiate the three lineages in separate clusters, with high level of confidence. As future work, the methodology will be assayed in commercial honey samples. ; info:eu-repo/semantics/publishedVersion

Unmanaged honey bee colonies of local ecotype surviving without human intervention are likely to form a valuable genetic resource for the sustainability of managed apiaries as well conservation of threatened subspecies. In Ireland, following the Isle of Wight disease (which devastated honey bee colonies at the beginning of the 20th century) and subsequent hybridisation with C lineage bees, there has been a general acceptance by government agencies, scientists, and many beekeepers that no Apis mellifera mellifera (Amm) colonies persisted in the wild. However, sporadic reports were received in 2014/2015 of the existence of unmanaged honey bee colonies. Given that Ireland's human population is low in density with only 32 persons per square km in some rural areas and only approximately 3000 registered beekeepers, many of whom are reported to not favour purchasing imported bees, it is feasible that honeybees could have naturally adapted to introduced pathogens such as Varroa destructor. We initiated an investigation into the state of unmanaged honey bee colonies and in 2016 we launched a nationwide request through press and social media seeking locations of unmanaged colonies which realised over 170 replies in a short time period. We found that unmanaged colonies have utilised a wide variety of both natural and artificial cavities and survived unaided for periods reported to be from three to over 20 years. Given the difficulty in confirming the authenticity of these timings the survival of individual colonies has been monitored since 2016. Sixty-two of the colonies were sampled and a combined approach using mitochondrial, microsatellite and single nucleotide polymorphism (SNP) genotyping has shown the majority to be pure Apis mellifera mellifera and forming an integral part of the previously described pure Amm population in Ireland. This data, along with survival records for >2 years, and details of surrounding habitat and health of the unmanaged colonies, will be presented. ; info:eu-repo/semantics/publishedVersion

With a growing number of parasites and pathogens experiencing large-scale range expansions, monitoring diversity in immune genes of host populations has never been so important because it can inform on the adaptive potential to resist the invaders. Population surveys of immune genes are becoming common in many organisms, yet they are missing in the honey bee (Apis mellifera L.), a key managed pollinator species that has been severely affected by biological invasions. To fill the gap, here we identified single nucleotide polymorphisms (SNPs) in a wide range of honey bee immune genes and developed a medium-density assay targeting a subset of these genes. Using a discovery panel of 123 whole-genomes, representing seven A. mellifera subspecies and three evolutionary lineages, 180 immune genes were scanned for SNPs in exons, introns (< 4 bp from exons), 3' and 5´UTR, and < 1 kb upstream of the transcription start site. After application of multiple filtering criteria and validation, the final medium-density assay combines 91 quality-proved functional SNPs marking 89 innate immune genes and these can be readily typed using the high-sample-throughput iPLEX MassARRAY system. This medium-density-SNP assay was applied to 156 samples from four countries and the admixture analysis clustered the samples according to their lineage and subspecies, suggesting that honey bee ancestry can be delineated from functional variation. In addition to allowing analysis of immunogenetic variation, this newly-developed SNP assay can be used for inferring genetic structure and admixture in the honey bee. ; We are deeply indebted to Frank Aguiar, Luís Silva, Edgardo Melo, João Martins, João Melo, Manuel Moura, Manuel Viveiros, and Ricardo Sousa from "Direção Regional da Agricultura e Desenvolvimento Rural dos Açores" (Portugal), and to Laura Garreau, Laurent Maugis, Pascale Sauvage and Jacques Kermagoret, from "Association Conservatoire de l'Abeille Noir Bretonne" (France), for sampling the apiaries in São Miguel, Santa Maria, and Ouessant islands. Genotyping was outsourced to the Epigenetics and Genotyping laboratory, Central Unit for Research in Medicine (UCIM), University of Valencia, Spain. Data analyses were performed using computational resources at the Research Centre in Digitalization and Intelligent Robotics (CeDRI), Instituto Politécnico de Bragança. Ana Rita Lopes is supported by a PhD scholarship (SFRH/BD/143627/2019) from the Foundation for Science and Technology (FCT), Portugal. FCT provided financial support by national funds (FCT/MCTES) to CIMO (UIDB/00690/2020).This research was funded through the projects BEEHAPPY (POCI-01-0145- FEDER-029871, FCT and COMPETE/QREN/EU) and BEEHEAL. BEEHEAL was funded by the ARIMNet2 2016 Call by the following agencies: INIA (Spain), MOARD (Israel), ANR (France) and FCT (Portugal). ARIMNet2 (ERA-NET) received funding from the European Union's Seventh Framework Programme for research, technological development and demonstration under grant agreement no. 618127. ; info:eu-repo/semantics/publishedVersion

In this study, we gathered sequence data from the tRNAleu-cox2 intergenic mitochondrial (mtDNA) region concurrently with single nucleotide polymorphism (SNP) data from 91 loci of nuclear DNA (ncDNA). The data was obtained from 156 colonies sampled in six apiaries from four countries. The full dataset was analysed and discussed for genetic patterns with a focus on cytonuclear diversity and admixture levels. ; This research was funded through the projects BEEHAPPY (POCI-01-0145-FEDER-029871, FCT and COMPETE/QREN/EU) and BEEHEAL. BEEHEAL was funded by the ARIMNet2 2016 Call by the following agencies: INIA (Spain), MOARD (Israel), ANR (France) and FCT (Portugal). ARIMNet2 (ERANET) received funding from the European Union's Seventh Framework Programme for research, technological development and demonstration under grant agreement no. 618127. Ana Rita Lopes is supported by a PhD scholarship (SFRH/BD/143627/2019) from the Foundation for Science and Technology (FCT), Portugal. FCT provided financial support by national funds (FCT/MCTES) to CIMO (UIDB/00690/2020). ; info:eu-repo/semantics/publishedVersion