Abstract
Background
Marker-assisted selection is well established in animal breeding method of selecting individuals with desirable traits in a breeding scheme based on DNA molecular marker patterns.
Objective
Genetic diversity and C-derived admixture into local purebred gene pool of A. m. mellifera colonies was assessed using polymorphism of nine microsatellite loci in order to provide further marker-assisted selection of desired honey bee colonies.
Methods
The genetic diversity and the level of C-derived introgression into A. m. mellifera colonies in the Shulgan-Tash Nature Reserve (Russia) was assessed based on nine microsatellite loci (ap243, 4a110, A24, A8, A43, A113, A88, Ap049, A28), which were analized using the fragment analysis of the PCR products in Applied Biosystems 3130 DNA Analyzer. Phylogenetic relationship of colonies was evaluated using Neighbor-Joining methods with Cavalli-Sforza and Edwards genetic distance using the PHYLIP 3.68. The model-based Bayesian clustering algorithm implemented in STRUCTURE 2.3.3 was employed to infer membership and introgression proportions (Q-value).
Results
In the Shulgan-Tash Nature Reserve colonies of A. m. mellifera subdivided into four groups by level of C-derived introgression. Only five colonies of A. m. mellifera had C-derived introgression which varied from 0.5 to 2%. The genetic diversity in colonies of A. m. mellifera varied from 0.12 to 0.40. The Neighbor-Joining tree demonstrates the genetic relationship of A. m. mellifera colonies, which subdivided into three groups with different levels of C-derived introgression. Group 1 combined five honey bee colonies Bort_1, Bort_2, Bort_3, Baisalyan_1, and Kush_7 with a fraction of introgression close to 0.000 and genetic diversity from 0.20 to 0.25.
Conclusion
The results showed the excellence of nine microsatellite loci genotyping in estimation of genetic diversity, distinguishing the two European evolutionary lineages M and C and estimating C-derived introgression. These genetic parameters can be applied further to perform the marker-assisted selection of purebred dark European honey bees.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Adam B (1983) In search of the best strains of bees. Northern Bee Books, Hebden Bridge
Allendorf FW, Luikart G, Aitken SN (2012) Conservation and the genetics of populations, 2nd edn. Wiley-Blackwell, Oxford
Alpatov VV (1948) Honeybee species and their use in agriculture. Moscow Society of Naturalists, Moscow
Anderson E (1968) Introgressive hybridization. vol ix. Biological research series, vol ix. Hafner Publishing Company, New York
Baer B, Schmid-Hempel P (1999) Experimental variation in polyandry affects parasite loads and fitness in a bumble-bee. Nature 397:151–154. https://doi.org/10.1038/16451
Baudry E, Solignac M, Garnery L, Gries M, Cornuet J, Koeniger N (1998) Relatedness among honeybees (Apis mellifera) of a drone congregation. Proc R Soc Lond B 265:2009–2014. https://doi.org/10.1098/rspb.1998.0533
Bertrand B, Alburaki M, Legout H, Moulin S, Mougel F, Garnery L (2015) MtDNA COI-COII marker and drone congregation area: an efficient method to establish and monitor honeybee (Apis mellifera L.) conservation centres. Mol Ecol Resour 15:673–683. https://doi.org/10.1111/1755-0998.12339
Bouga M, Harizanis PC, Kilias G, Alahiotis S (2005) Genetic divergence and phylogenetic relationships of honey bee Apis mellifera (Hymenoptera: Apidae) populations from Greece and Cyprus using PCR-RFLP analysis of three mtDNA segments. Apidologie 36:335–344. https://doi.org/10.1051/apido:2005021
Büchler R, Costa C, Hatjina F, Andonov S, Meixner MD, Conte YL, Uzunov A, Berg S, Bienkowska M, Bouga M et al (2014) The influence of genetic origin and its interaction with environmental effects on the survival of Apis mellifera L. colonies in Europe. J Apic Res 53:205–214. https://doi.org/10.3896/IBRA.1.53.2.03
Cavalli-Sforza LL, Edwards AWF (1967) Phylogenetic analysis: models and estimation procedures. Am J Hum Genet 19:233–257
De la Rúa P, Jaffé R, Dall ‘olio R, Noz IM, Serrano JE (2009) Biodiversity, conservation and current threats to European honeybees*. Apidologie 40:263–284. https://doi.org/10.1051/apido/2009027
De la Rúa P, Jaffé R, Muñoz I, Serrano J, Moritz RF, Kraus FB (2013) Conserving genetic diversity in the honeybee: comments on Harpur et al. (2012). Mol Ecol 22:3208–3210. https://doi.org/10.1111/mec.12333
Dietemann V, Pirk CWW, Crewe R (2009) Is there a need for conservation of honeybees in Africa? Apidologie 40:285–295. https://doi.org/10.1051/apido/2009013
Earl DA, Vonholdt BM (2012) Structure harvester: a website and program for visualizing structure output and implementing the Evanno method. Conserv Genet Resour 4:359–361. https://doi.org/10.1007/s12686-011-9548-7
Engel MS (1999) The taxonomy of recent and fossil honey bees (Hymenoptera, Apidae, Apis). J Hymenoptera Res 8:165–196. https://doi.org/10.1007/978-1-4614-4960-718
Estoup A, Garnery L, Solignac M, Cornuet J-M (1995) Microsatellite variation in honey bee (Apis mellifera L.) populations: hierarchical genetic structure and test of the infinite allele and stepwise mutation models. Genetics 140:679–695. https://doi.org/10.1080/00218839.1999.11100990
Evanno G, Regnaut S, Goudet J (2005) Detecting the number of clusters of individuals using the software STRUCTURE: a simulation study. Mol Ecol 14:2611–2620. https://doi.org/10.1111/j.1365-294X.2005.02553.x
Felsenstein J (1993) PHYLIP (phylogeny inference package), version 3.5c. University of Washington, Seattle
Frankham R, Ballou JD, Briscoe DA (2002) Introduction to conservation genetics. Cambridge University Press, Cambridge
Fuchs S, Moritz RFA (1999) Evolution of extreme polyandry in the honeybee Apis mellifera L. Behav Ecol Sociobiol 45:269–275. https://doi.org/10.1007/s002650050561
Fuchs S, Schade V (1994) Lower performance in honeybee colonies of uniform paternity. Apidologie 25:155–168. https://doi.org/10.1051/apido:19940204
Gallai N, Salles JM, Settele J, Vaissiere BE (2009) Economic valuation of the vulnerability of world agriculture confronted with pollinator decline. Ecol Econ 68:810–821. https://doi.org/10.1016/j.ecolecon.2008.06.014
Garnery L, Solignac M, Celebrano G, Cornuet J-M (1993) A simple test using restricted PCR-amplified mitochondrial DNA to study the genetic-structure of Apis mellifera L. Cell Mol Life Sci 49:1016–1021. https://doi.org/10.1007/BF02125651
Goudet J (1995) FSTAT (version 1.2): a computer program to calculate F-statistics. J Hered 86:485–486. https://doi.org/10.1093/oxfordjournals.jhered.a111627
Haberl M, Tautz D (1999) Tri- and tetranucleotide microsatellite loci in honey bees (Apis mellifera)—a step towards quantitative genotyping. Mol Ecol 8:1358–1360. https://doi.org/10.1046/j.1365-294X.1999.00701_5.x
Hall MA, Jones J, Rocchetti M, Wright D, Rader R (2020) Bee visitation and fruit quality in berries under protected cropping vary along the length of polytunnels. J Econ Entomol 113:toaa037. https://doi.org/10.1093/jee/toaa037
Harpur BA, Minaei S, Kent CF, Zayed A (2012) Management increases genetic diversity of honey bees via admixture. Mol Ecol 21:4414–4421. https://doi.org/10.1111/j.1365-294X.2012.05614.x
Harrison RG (1993) Hybrid zones and the evolutionary process. Oxford University Press, Oxford. ISBN 0-19-506917-X
Harrison RG, Larson EL (2014) Hybridization, introgression, and the nature of species boundaries. J Hered 105:795–809. https://doi.org/10.1093/jhered/esu033
Henriques D, Parejo M, Vignal A, Wragg D, Wallberg A, Webster MT, Pinto MA (2018) Developing reduced SNP assays from whole-genome sequence data to estimate introgression in an organism with complex genetic patterns, the Iberian honeybee (Apis mellifera iberiensis). Evol Appl 11:1270–1282. https://doi.org/10.1111/eva.12623
Hepburn HR, Radloff SE, Fuchs S (1998) Population structure and the interface between Apis mellifera capensis and Apis mellifera scutellata. Apidologie 29:333–346. https://doi.org/10.1051/apido:19980404
Ilyasov RA, Petukhov AV, Poskryakov AV, Nikolenko AG (2007) Local honeybee (Apis mellifera mellifera L.) populations in the Urals. Russ J Genet 43:709–711. https://doi.org/10.1134/S1022795407060166
Ilyasov RA, Poskryakov AV, Petukhov AV, Nikolenko AG (2015) Genetic differentiation of local populations of the dark European bee Apis mellifera mellifera L. in the Urals. Russ J Genet 51:677–682. https://doi.org/10.1134/S1022795415070042
Ilyasov RA, Poskryakov AV, Petukhov AV, Nikolenko AG (2016) Molecular genetic analysis of five extant reserves of black honeybee Apis mellifera mellifera in the Urals and the Volga region. Russ J Genet 52:828–839. https://doi.org/10.1134/S1022795416060053
Ilyasov RA, Poskryakov AV, Nikolenko AG (2017) Modern methods of assessing the taxonomic affiliation of honeybee colonies. Ecol Genet 15:41–51. https://doi.org/10.17816/ecogen15441-51
Jensen AB, Palmer KA, Boomsma JJ, Pedersen BV (2005) Varying degrees of Apis mellifera ligustica introgression in protected populations of the black honeybee, Apis mellifera mellifera, in northwest Europe. Mol Ecol 14:93–106. https://doi.org/10.1111/j.1365-294X.2004.02399.x
Klein A-M, Vaissière BE, Cane JH, Steffan-Dewenter I, Cunningham SA, Kremen C, Tscharntke T (2007) Importance of pollinators in changing landscapes for world crops. Proc R Soc B Biol Sci 274:303–313. https://doi.org/10.1098/rspb.2006.3721
Liersch S, Schmid-Hempel P (1998) Genetic variation within social insect colonies reduces parasite load. Proc R Soc Lond B 265:221–225. https://doi.org/10.1098/rspb.1998.0285
Meixner MD, Costa C, Kryger P, Hatjina F, Bouga M, Ivanova E, Büchler R (2010) Conserving diversity and vitality for honey bee breeding. J Apic Res 49:85–92. https://doi.org/10.3896/IBRA.1.49.1.12
Meixner MD, Leta MA, Koeniger N, Fuchs S (2011) The honey bees of Ethiopia represent a new subspecies of Apis mellifera—Apis mellifera simensis n. ssp. Apidologie 42:425–437. https://doi.org/10.1007/s13592-011-0007-y
Moritz RFA, Neumann P (2010) Genetic analysis of the drifting of drones in apis mellifera using multilocus DNA fingerprinting. Ethology 102:580–590. https://doi.org/10.1111/j.1439-0310.1996.tb01150.x
Muñoz I, De la Rúa P (2012) Temporal analysis of the genetic diversity in a honey bee mating area of an island population (La Palma, Canary Islands, Spain). J Apicult Sci 56:141–148. https://doi.org/10.2478/v10289-012-0005-y
Muñoz I, Stevanovic J, Stanimirovic Z, De la Rúa P (2012) Genetic variation of Apis mellifera from Serbia inferred from mitochondrial analysis. J Apicult Sci 56:59–69. https://doi.org/10.2478/v10289-012-0007-9
Muñoz I, Pinto MA, De la Rúa P (2014) Effects of queen importation on the genetic diversity of Macaronesian island honey bee populations (Apis mellifera Linneaus 1758). J Apic Res 53:296–302. https://doi.org/10.3896/IBRA.1.53.2.11
Muñoz I, Henriques D, Johnston JS, Chávez-Galarza J, Kryger P, Pinto MA (2015) Reduced SNP panels for genetic identification and introgression analysis in the dark honey bee (apis mellifera mellifera). plos One 10:e0124365. https://doi.org/10.1371/journal.pone.0124365
Muñoz I, Henriques D, Jara L, Johnston JS, Chávez-Galarza J, De La Rúa P, Pinto MA (2017) SNPs selected by information content outperform randomly selected microsatellite loci for delineating genetic identification and introgression in the endangered dark European honeybee (Apis mellifera mellifera). Mol Ecol Resour 17:783–795. https://doi.org/10.1111/1755-0998.12637
Nedić N, Francis RM, Stanisavljević L, Pihler I, Kezić N, Bendixen C, Kryger P (2014) Detecting population admixture in honey bees of Serbia. J Apic Res 53:303–313. https://doi.org/10.3896/IBRA.1.53.2.12
Nelson R, Wallberg A, Simões Z, Lawson D, Webster M (2017) Genomewide analysis of admixture and adaptation in the Africanized honeybee. Mol Ecol 26:3603–3617. https://doi.org/10.1111/mec.14122
Neumann P, Carreck NL (2010) Honey bee colony losses. J Apic Res 49:1–6. https://doi.org/10.3896/IBRA.1.49.1.01
Neumann P, Moritz RFA, van Praagh J (1999) Queen mating frequency in different types of honey bee mating apiaries. J Apic Res 38:11–18. https://doi.org/10.1080/00218839.1999.11100990
Oldroyd B, Rinderer T, Buco S (1992) Intra-colonial foraging specialism by honey bees (Apis mellifera) (Hymenoptera: Apidae). Behav Ecol Sociobiol 30:291–295. https://doi.org/10.1007/BF00170594
Oleksa A, Chybicki I, Tofilski A, Burczyk J (2011) Nuclear and mitochondrial patterns of introgression into native dark bees (Apis mellifera mellifera) in Poland. J Apic Res 50:116–129. https://doi.org/10.3896/IBRA.1.50.2.03
Oleksa A, Wilde J, Tofilski A, Chybicki IJ (2013) Partial reproductive isolation between European subspecies of honey bees. Apidologie 44:611–619. https://doi.org/10.1007/s13592-013-0212-y
Oxley PR, Hinhumpatch P, Gloag R, Oldroyd BP (2010) Genetic evaluation of a novel system for controlled mating of the honeybee, Apis mellifera. J Hered 101:334–338. https://doi.org/10.1093/jhered/esp112
Page RE, Robinson GE, Fondrk MK, Nasr ME (1995) Effects of worker genotypic diversity on honey bee colony development and behavior (Apis mellifera L.). Behav Ecol Sociobiol 36:387–396. https://doi.org/10.1007/BF00177334
Palmer MR, Smith DR, Kaftanoğlu O (2000) Turkish honeybees: genetic variation and evidence for a fourth lineage of Apis mellifera mtDNA. J Hered 91:42–46. https://doi.org/10.1093/jhered/91.1.42
Parejo M, Henriques D, Pinto MA, Soland-Reckeweg G, Neuditschko M (2018) Empirical comparison of microsatellite and SNP markers to estimate introgression in Apis mellifera mellifera. J Apic Res 57:504–506. https://doi.org/10.1080/00218839.2018.1494894
Péntek-Zakar E, Oleksa A, Borowik T, Kusza S (2015) Population structure of honey bees in the Carpathian Basin (Hungary) confirms introgression from surrounding subspecies. Ecol Evol 5:5456–5467. https://doi.org/10.1002/ece3.1781
Pinto MA, Henriques D, Chávez-Galarza J, Kryger P, Garnery L, van der Zee R, Dahle B, Soland-Reckeweg G, De la Rúa P, Dall’ Olio R et al (2014) Genetic integrity of the dark European honey bee (Apis mellifera mellifera) from protected populations: a genome-wide assessment using SNPs and mtDNA sequence data. J Apic Res 53:269–278. https://doi.org/10.3896/IBRA.1.53.2.08
Pritchard JK, Stephens M, Donnelly P (2000) Inference of population structure using multilocus genotype data. Genetics 155:945–959
Raymond M, Rousset F (1995) GENEPOP (Version 1.2): population genetics software for exact tests and ecumenicism. J Hered 86:248–249. https://doi.org/10.1093/oxfordjournals.jhered.a111573
Ruttner F (1988) Biogeography and taxonomy of honeybees. Springer, Berlin, p 288. https://doi.org/10.1016/0169-5347(89)90176-6
Scharpenberg H, Neumann P, van Praagh JP, Moritz RFA (2006) Reliability of an island mating apiary under routine management. J Apic Res 45:153–154. https://doi.org/10.1080/00218839.2006.11101334
Schneider SS, DeGrandi-Hoffman G, Smith DR (2004) The African honeybee: factors contributing to a successful biological invasion. Annu Rev Entomol 49:351–376. https://doi.org/10.1146/annurev.ento.49.061802.123359
Seeley TD, Visscher PK (1985) Survival of honeybees in cold climates: the critical timing of colony growth and reproduction. Ecol Entomol 10:81–88. https://doi.org/10.1111/j.1365-2311.1985.tb00537.x
Seeley TD, Tarpy DR, Griffin SR, Carcione A, Delaney DA (2015) A survivor population of wild colonies of European honeybees in the northeastern United States: investigating its genetic structure. Apidologie 46:654–666. https://doi.org/10.1007/s13592-015-0355-0
Sheppard WS, Meixner MD (2003) Apis mellifera pomonella, a new honey bee subspecies from Central Asia. Apidologie 34:367–375. https://doi.org/10.1051/apido:2003037
Shykoff JA, Schmid-Hempel P (1991) Parasites and the advantage of genetic variability within social insect colonies. Proc R Soc Lond B 243:55–58. https://doi.org/10.1098/rspb.1991.0009
Soland-Reckeweg G, Heckel G, Neumann P, Fluri P, Excoffier L (2009) Gene flow in admixed populations and implications for the conservation of the Western honeybee, Apis mellifera. J Insect Conserv 13:317–328. https://doi.org/10.1007/s10841-008-9175-0
Solignac M, Vautrin D, Loiseau A, Mougel F, Baudry E, Estoup A, Garnery L, Haberl M, Cornuet JM (2003) Five hundred and fifty microsatellite markers for the study of the honeybee (Apis mellifera L.) genome. Mol Ecol Notes 3:307–311. https://doi.org/10.1046/j.1471-8286.2003.00436.x
Southwick EE, Southwick L (1992) Estimating the economic value of honey bees (Hymenoptera: Apidae) as agricultural pollinators in the United States. J Econ Entomol 85:621–633. https://doi.org/10.1093/jee/85.3.621
Stevanovic J, Stanimirovic Z, Radakovic M, Kovacevic SR (2010) Biogeographic study of the honey bee (Apis mellifera L.) from Serbia, Bosnia and Herzegovina and Republic of Macedonia Based on mitochondrial DNA analyses. Russ J Genet 46:603–609. https://doi.org/10.1134/S1022795410050145
Strange JP, Garnery L, Sheppard WS (2008) Morphological and molecular characterization of the Landes honey bee (Apis mellifera L.) ecotype for genetic conservation. J Insect Conserv 12:527–537. https://doi.org/10.1007/s10841-007-9093-6
Uzunov A, Kiprijanovska H, Andonov S, Naumovski M, Gregorc A (2009) Morphological diversity and racial determination of the honey bee (Apis mellifera L.) population in the Republic of Macedonia. J Apic Res 48:196–203. https://doi.org/10.3896/IBRA.1.48.3.08
Uzunov A, Meixner MD, Kiprijanovska H, Andonov S, Gregorc A, Ivanova E, Bouga M, Dobi P, Büchler R, Francis R et al (2014) Genetic structure of apis mellifera macedonica in the Balkan peninsula based on microsatellite DNA polymorphism. J Apic Res 53:288–295. https://doi.org/10.3896/IBRA.1.53.2.10
Wallberg A, Bunikis I, Pettersson OV, Mosbech M-B, Childers AK, Evans JD, Mikheyev AS, Robertson HM, Robinson GE, Webster MT (2019) A hybrid de novo genome assembly of the honeybee, Apis mellifera, with chromosome-length scaffolds. BMC Genom 20:275. https://doi.org/10.1186/s12864-019-5642-0
Woyciechowski M, Warakomska Z (1994) Worker’s genetic diversity has no relation to pollen diversity in a honey bee colony (Apis mellifera L.). J Ethol 12:163–167. https://doi.org/10.1007/BF02350060
Acknowledgements
This work was supported by postdoctoral fellowships from Incheon National University to RI (2017).
Author information
Authors and Affiliations
Contributions
All authors contributed to the study conception and design. Material preparation, data collection, and analysis were performed by RI, MLL, UY, and AN. The first draft of the manuscript was written by RI and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.
Corresponding authors
Ethics declarations
Conflict of interest
Rustem Ilyasov, Myeong-Lyeol Lee, Ural Yunusbaev, Alexey Nikolenko, and Hyung-Wook Kwon declare that they do not have a conflict of interest.
Informed consent
The authors’ consent to publish and a copyright transfer.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Ilyasov, R.A., Lee, ML., Yunusbaev, U. et al. Estimation of C-derived introgression into A. m. mellifera colonies in the Russian Urals using microsatellite genotyping. Genes Genom 42, 987–996 (2020). https://doi.org/10.1007/s13258-020-00966-0
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s13258-020-00966-0