Abstract
The genetic variability of Apis mellifera ligustica was screened throughout the Italian peninsula and Sardinia with eight polymorphic microsatellite loci. Samples of Apis mellifera mellifera, Apis mellifera carnica and from the Buckfast breeding line were genotyped for comparison. Low Fis and Fst values suggest the absence of genetic structure within and among A. m. ligustica populations, although the high number of alleles detected and heterozygosity. Phylogenetic and individual analyses confirmed that A. m. ligustica has come to resemble one large population, probably as a result of intensive beekeeping practices such as migratory beekeeping and large-scale commercial queen trading. Since the introgression of foreign alleles into both endemic natural and commercial A. m. ligustica populations, can be detected and monitored by microsatellite analysis, the results provide a reference data set for future local biodiversity conservation and other controlled breeding programs.
Zusammenfassung
Wir erfassten die genetische Variabilität von Honigbienen der Rasse Apis mellifera ligustica über ganz Italien hinweg und in Sardinien. Die Genotypen zweier anderer Unterarten (Apis mellifera mellifera, Apis mellifera carnica) und Proben der künstlichen Buckfast-Zuchtlinie wurden zum Vergleich ebenfalls untersucht. Insgesamt wurden über vier Jahre hinweg (2001–2004) Proben aus 379 Völkern von verschiedenen Standorten genommen. Alle A. m. ligustica Proben wurden sieben verschiedenen regionalen Gruppen zugeordnet (Abb. 1). Genetische Polymorphismen wurden für acht Mikrosatellitenloci mittels automatisierter Kapillarelektrophorese untersucht (Locus-spezifische PCR-Bedingungen sind in Tab. I zusammengestellt). Die Mikrosatellitendaten der zehn Gruppen ermöglichte die Erfassung der allgemeinen Populationsparameter (Tab. II), die Darstellung der phylogenetischen Beziehungen zwischen den Gruppen und deren jeweilige Populationsstruktur, ebenso wie die Erstellung individueller Analysen.
Die in dieser Studie verwendeten genetischen Marker belegen das Potential der Mikrosatelliten-DNA-Loci für die Unterscheidung eng verwandter Taxa, da es möglich war, nicht nur A. m. mellifera- Proben (dem Evolutionszweigs M zugeordnet) von anderen zu unterscheiden, sondern auch zwischen A. m. carnica and A. m. ligustica (beide zum gleichen geographischen Zweig C gehörend) zu differenzieren. Die meisten Loci hatten unterartspezifische Allele (Tab. III), wobei der Locus Ap43 ein jeweils exklusives Allel für jedes der drei Taxa aufwies. Die Analyse der künstlichen Buckfast-Zuchtlinie belegte deren Ursprung als Hybrid und deren hohe genetische Diversität. Unser Genotypisierungsansatz erlaubte zusätzlich die Detektion von Hybridindividuen in Bienenproben aus dem Nordosten Italiens (Abb. 2).
Phylogenetische und individuelle Analysen belegten, dass A. m. ligustica inzwischen eine grosse Population darstellt, wahrscheinlich als Ergebnis einer intensiven Bienenhaltung, insbesondere durch die Wanderung von Bienenvölkern und den grossangelegten kommerziellen Austausch von Königinnen. Die Ergebnisse unserer Untersuchung stellen ein Datenset dar, das künftigen Programmen zur lokalen Konservierung der Biodiversität und auch kontrollierten Zuchtprogrammen von Nutzen sein kann. Die Mikrosatelliten-DNA-Analyse ist ein effizientes Verfahren, um die Introgression fremder Allele in natürlichen und kommerziell genutzten A. m. ligustica-Populationen zu entdecken und zu verfolgen.
References
Badino G., Celebrano G., Manino A. (1982) Genetic variability of Apis mellifera ligustica Spin. in a marginal area of its geographical distribution, Experientia 38, 540–541.
Badino G., Celebrano G., Manino A. (1983) Population structure and Mdh-1 locus variation in Apis mellifera ligustica, J. Hered. 74, 443–446.
Bar-Cohen R., Alpern G., Bar-Anan R. (1978) Progeny testing and selecting Italian queen for brood area and honey production, Apidologie 9, 95–100.
Bolchi Serini G., Sommaruga A., Lapietra G. (1983) Studio biometrico di popolazioni alpine di Apis mellifera L., Boll. Zool. Agrar. Bachic. 17, 1–18.
Cavalli-Sforza L., Edwards A.W.E. (1967) Phylogenetic analysis: models and estimation procedures, Am. J. Human Genet. 19, 233–257.
Comparini A., Biasiolo A. (1991) Genetic characterization of Italian bee Apis mellifera ligustica Spin, versus Carnolian bee, Apis mellifera carnica Poll, by allozyme variability analysis, Biochem. Syst. Ecol. 19, 189–194.
Cornuet J.M., Fresnaye J. (1989) Étude biométrique de colonies d’abeilles d’Espagne et du Portugal, Apidologie 20, 93–101.
Cornuet J.M., Daoudi A., Mossine E.H., Fresnate J. (1988) Étude biométrique de populations d’abeilles marocaines, Apidologie 19, 355–366.
Estoup A., Garnery L., Solignac M., Cornuet J.M. (1995) Microsatellite variation in honeybee (Apis mellifera L.) populations: hierarchical genetic structure and test of the infinite allele and stepwise mutation models, Genetics 140, 679–695.
Felsenstein J. (1993) PHYLIP, phytogeny inference package, Version 3.57, University of Washington, Seattle.
Floris I., Prota R. (1994) Variazioni di alcune caratteristiche morfometriche nella popolazione di Apis mellifera L. della Sardegna nell’ultimo ventennio, Apicoltura 9, 163–175.
Franck P. (2004) Genetic differentiation in a suture zone between Apis mellifera mellifera and A.m. ligustica subspecies in the Aosta valley, Proc. 1st Eur. Conf. Apidology, EurBee, Udine, Italy, 19–23 September.
Franck P., Garnery L., Solignac M., Cornuet J.M. (1998) The origin of west European subspecies of honeybees (Apis mellifera): new insights from microsatellite and mitochondrial data, Evolution 52, 1119–1134.
Franck P., Garnery L., Celebrano G., Solignac M., Cornuet J.M. (2000) Hybrid origin of honeybees from Italy (Apis mellifera ligustica) and Sicily (A. m. sicula), Mol. Ecol. 9, 907–921.
Garnery L., Cornuet J.M., Solignac M. (1992) Evolutionary history of the honey bee Apis mellifera inferred from mitochondrial DNA analysis, Mol. Ecol. 1, 145–154.
Garnery L., Franck P., Baudry E., Vautrin D., Cornuet J.M. Solignac M. (1998) Genetic biodiversity of the West European honeybee (Apis mellifera mellifera and Apis mellifera iberica). II. Microsatellite loci, Genet. Sel. Evol. 30, 49–74.
Goldstein D.B., Ruiz Linares A., Cavalli-Sforza L.L., Feldman M.W. (1995) Genetic absolute dating based on microsatellites and origin of modern humans, Proc. Natl. Acad. Sci. USA 92, 6723–6727.
Goudet J. (1995) FSTAT (version 1.2): A Computer Program to Calculate F-Statistics, J. Hered. 86, 485–486.
Hedges S.B. (1992) The number of replications needed for accurate estimation of the bootstrap P-value in phylogenetic studies, Mol. Biol. Evol. 9, 366–369.
Hewitt G.M. (1996) Some genetic consequences of ice ages, and their role in divergence and speciation, Biol. J. Linn. Soc. 58, 247–276.
Hewitt G.M. (1999) Post-glacial re-colonization of European biota, Biol. J. Linn. Soc. 68, 87–112.
Lebdigrissa K., Msadda K., Cornuet J.M., Fresnaye J. (1991) Phylogenetic relationship between the Tunisian honeybee Apis mellifera intermissa and neighbouring African and west Mediterranean honeybee breeds, Landbouwtijdschrift 44, 1231–1238.
Lodesani M., Colombo R., Spreafico C. (1995) Ineffectiveness of Apistan treatment against the mite Varroa jacobsoni Oud in several districts of Lombardy (Italy), Apidologie 26, 67–72.
Manino A., Marietta F. (1984) II sistema enzimatico MDH in popolazioni di Apis mellifera L. della Valle d’Aosta, L’Apicoltore Moderno 75, 89–94.
Marino A., Sabatini A.G., Carpana E., Mantovani B. (2002) Analysis of mitochondrial genes NDH2, CO1 and 16S for the characterization of Apis mellifera populations, Insects Social Life 4, 87–92.
Marietta F., Manino A., Pedrini P. (1984a) Intergradazione tra sottospecie di Apis mellifera in Liguria, L’Apicoltore Moderno 75, 159–163.
Marietta F., Manino A., Balboni G. (1984b) Indagini Biometriche su popolazioni di Apis mellifera L. delle Alpi occidentali, L’Apicoltore Moderno 75, 213–223.
Meixner M., Sheppard W.S., Poklukar J. (1993) Asymmetrical distribution of a mitochondrial DNA polymorphism between two introgressing honey bee races, Apidologie 24, 147–153.
Nazzi F. (1992) Morphometric analysis of honey bees from an area of racial hybridization in northeastern Italy, Apidologie 23, 89–96.
Nei M. (1978) Estimation of average heterozygosity and genetic distance from a small number of individuals, Genetics 89, 145–163.
Pritchard J.K., 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 test and ecumenism, J. Hered. 86, 248–250.
Ruttner F. (1988) Biogeography and Taxonomy of Honeybees, Springer Verlag, Berlin.
Saitou N., Nei M. (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees, Mol. Biol. Evol. 4, 406–425.
Sheppard W.S. (1989) A history of the introduction of honey bee races into the United States-I and II, Am. Bee J. 129, 617–619, 664–667.
Solignac M., Vautrin D., Loiseau A., Mougel F., Baudry E., Estoup A., Garnery L., Haberl M., Cornuet J.M. (2003) Five hundred and fifty microsatellite markers for the study of the honeybee (Apis mellifera L.) genome, Mol. Ecol. Notes 3, 307–311.
Taberlet P., Fumagalli L., Wust-Saucy A., Cosson J. (1998) Comparative phylogeography and post glacial colonization routes in Europe, Mol. Ecol. 7, 453–464.
Verardi A., Arduino P., Floris I., Prota R., Bullini L. (1998) Marcatori molecolari e introgressione dell’ape (Apis mellifera L.) in Sardegna, Proc. XVIII Ital. Natl. Congr. Entomol., 200.
Walsh P.S., Metzqer D.A., Higuchi R. (1991) Chelex 100 as a medium for simple extraction of DNA for PCR-based typing from forensic material, Biotechniques 10, 506–512.
Woodward D. (1993) Ligurian bees, Am. Bee J. 133, 124–125.
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Dall’Olio, R., Marino, A., Lodesani, M. et al. Genetic characterization of Italian honeybees, Apis mellifera ligustica, based on microsatellite DNA polymorphisms. Apidologie 38, 207–217 (2007). https://doi.org/10.1051/apido:2006073
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DOI: https://doi.org/10.1051/apido:2006073