Genetic and morphometric variation in honeybee (Apis mellifera L.) populations of Turkey

Apidologie, Jul 2018

Irfan Kandemir, Meral Kence, Aykut Kence

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Genetic and morphometric variation in honeybee (Apis mellifera L.) populations of Turkey

Apidologie Genetic and morphometric variation in honeybee (Apis mellifera L.) populations of Turkey n KANDEMIR 0 l KENCE 0 Aykut KENCE 0 0 Department of Biology, Middle East Technical University , 06531 Ankara , Turkey - Six enzyme systems were studied to determine the genetic variability in honeybee populations in Turkey. Ten morphometric characters were also measured to determine the extent of morphometric variation. Out of six enzyme systems, four were found to be polymorphic with 16 allozymes. The average heterozygosity was calculated as 0.072 ± 0.007. Morphometric and electrophoretic variables were equally effective in discriminating honeybee populations. European and Anatolian honeybees were separated on the first axis, and Anatolian honeybees were further separated along a second canonical axis. The observation of rare alleles in isoenzymes, detection of high genetic diversity and the presence of four known subspecies support the argument that Anatolia has been a genetic center for honeybee populations in the Near East. 1. INTRODUCTION Ruttner [ 30 ] claimed that southwest Asia is a zone of high morphological diversification and evolution for honeybees. Many clearly distinct races have evolved within this region, which includes a diversity of habitats. Asia Minor, including Anatolia, appears to be the genetic center for these honeybee subspecies according to the multivariate statistical analysis of morphometric data [ 30 ]. Honey bee races in this region include the subspecies Apis mellifera anatoliaca, A. m. caucasica, A. m. meda, and A. m. syriaca, which were considered by Ruttner [ 30 ] to form a basal branch (O) of the species. Another subspecies that is found in the European part of Turkey, i.e., Thrace, may be A. m. carnica, which belongs to the branch C of Ruttner’s classification. Migratory beekeeping has become widespread in Turkey within the last 20–30 years. Thousands of colonies are overwintered in the Mediterranean and Aegean regions, and then moved to central and eastern Anatolia during the summer and fall. These practices might promote the gene flow between different races, and result in homogenization of the gene pool of Anatolian honeybees. Despite the apparent importance of Anatolia in the evolution of honeybees, very little work has been done on the morphological and genetic diversity of Anatolian honeybees [ 14, 37 ]. In this study, we aimed to determine the extent of morphometric and genetic variation of honeybees distributed widely across Turkey. Ten morphometric variables were measured, and electrophoretic variation was studied in six enzyme systems. 2. MATERIALS AND METHODS Honeybee samples were collected in 1994–1996 between March and September in Turkey. Samples were taken from 77 different locations in 36 provinces from different geographic regions of Turkey. Turkey is divided into seven geographic regions differing both in climatic conditions and in geological structure. Sampling was carried out mostly from small apiaries which do not practice migratory beekeeping, and the hives sampled were stationary during the MarchSeptember sampling period. Requeening of colonies was mostly natural, although some beekeepers reported that occasionally queens had been purchased for some colonies. In all cases we attempted to sample colonies that had no history of management for requeening. Special care was taken to sample from localities that were not frequented by migratory beekeepers. Approximately 3 000 worker bees were collected, and were put into small plastic bottles, which were labeled; the insects were fed either with honey cake (honey and powdered sugar [1:1]) or with ‘Turkish delight’ (water + saccharose + starch), and brought live to the laboratory. Honeybees were dissected, the thoraces were ground, and the homogenates were kept frozen until needed for electrophoresis. Forewings and hind legs were mounted on a microscope slide for morphometric analysis. Microscope slides of legs and wings were projected onto a TV screen, and measurements were taken. In the present study, ten morphometric characters were measured, i.e., four for the hind legs, four for the forewings (according to Ruttner [ 30 ]), and an additional two forewing characters, distance c and distance d as determined by Nazzi [ 20 ]. Six enzyme systems (esterase: 3.1.1.1; hexokinase: 2.7.1.1; malate dehydrogenase: 1.1.1.37; malic enzyme: 1.1.1.40; phosphoglucomutase: 2.7.5.1; and phosphoglucose isomerase: 5.3.1.9), known to be polymorphic in A. mellifera, were utilized as biochemical markers. Starch-gel electrophoresis, gel and sample preparation and experimental conditions have been reported previously [ 14 ]. All allozymes were designated by using relative mobilities, with the most common allozyme used as standard (relative mobility: 100). Gene frequencies, enzyme heterozygosities and population heterozygosities were calculated according to Nei [ 22 ], using BIOSYS [ 39 ]. Goodnessof-fit of genotypic frequencies according to Hardy–Weinberg expectations were tested by the c 2-test [ 38 ]. Multivariate statistical analyses were applied to both morphometric and electrophoretic data using SYN-TAX V [ 28 ]. A phenogram of samples from seven geographic regions was constructed using the Mahalanobis distances among centroids of groups in discriminant function by UPGMA in NTSYS-PC 1.70 [ 29 ]. Regressions of morphometric and electrophoretic variables on latitude and longitude were computed using SYSTAT-7.0 [ 19 ]. 3. RESULTS 3.2. Est-3 locus The mean values of the characters measured and standard errors have been shown in Table I, together with the number of hives and the total number of individuals in each province. Analysis of variance (ANOVA) of the data showed a high heterogeneity among honeybee populations. Out of 19 variables (10 morphometric and 9 electrophoretic), 11 displayed significant heterogeneity. Mdh65 and Mdh100 gene frequencies and distance d, wing length variables were found to be highly heterogeneous (P < 0.001). Out of the six enzyme systems assayed, four were found to be polymorphic and two exhibited invariant banding patterns (Tab. II, and Figs. 1–4). The populations of honeybees in Turkey were found to be in Hardy-Weinberg equilibrium with respect to all polymorphic enzymes, except the Pgm enzyme system in the majority of southern honeybee populations, where deviations were in favor of heterozygotes. Out of 77 sampling localities, in 26 of these, there were significant deviations in favor of Pgm heterozygotes (17 localities; P < 0.001; 2 localities; P < 0.01; 7 localities; P < 0.05). 3.1. Pgm locus The Pgm locus exhibited four alleles, Pgm45, Pgm63, Pgm75 and Pgm100 according to their relative mobilities in the present study. The frequency of the most common allele (Pgm75) ranged between 0.500–0.976 in 35 polymorphic locations. A significant linear relationship was revealed by the regression of the Pgm allele (Pgm75 and Pgm100) frequencies on latitude. This is the first report of such a relationship published in the literature. The distribution of allele frequencies where the rare alleles are pooled is given in Figure 1. The Est-3 locus exhibited three alleles, Est70, Est100, Est130, as reported previously for Czechoslovakian honeybees by Sheppard and McPheron [ 35 ], and in central Anatolian honeybees by Kandemir and Kence [ 14 ]. These alleles correspond to EstS, EstM, and EstF respectively, in A. m. ligustica [2] and in Greek honeybees [3]. The frequency of the most common allele at the 22 polymorphic locations ranged between 0.853 and 0.995. Esterase was fixed for the Est100 allele in 14 locations in the northern and eastern provinces. Generally, there was a north to south differentiation in esterase allele frequencies. This conclusion was also confirmed by a significant linear relationship between the frequency of Est70 and latitude. The distribution of allele frequencies where the rare alleles are pooled is given in Figure 2. 3.3. Hk locus In the Hk locus, five alleles (Hk77, Hk87, Hk100, Hk110 and Hk120) were found in different honeybee populations in Turkey. The frequency of the most common allele in the 19 locations where this enzyme was polymorphic ranged between 0.707–0.990. In Thrace, samples were monomorphic for the Hk locus. The majority of samples taken from the Black Sea region were fixed for the Hk100 allele, whereas hexokinase was polymorphic in southern Anatolia. There was also a north to south differentiation in the hexokinase allele frequencies, shown by a significant linear relationship between Hk110 gene frequencies and latitude. The distribution of allele frequencies where the rare alleles are pooled is shown in Figure 4. 3.4. Mdh locus Four alleles (Mdh65, Mdh87, Mdh100 and Mdh116) were found for this enzyme in the m .fooN seeb m h t i r 0 6 2 0 0 6 4 6 3 5 8 3 0 4 4 3 2 9 6 6 4 5 7 1 2 9 2 5 5 3 7 8 1 0 2 3 isn .00± .00± .00± .00± .00± .00± .00± .00± .00± .00± .00± .00± .00± .00± .00± .00± .00± .00± .00± .00± .00± .00± .00± .00± .00± .00± .00± .00± .00± .00± .00± .00± .00± .00± .00± .00± 2 3 6 2 1 1 1 1 1 1 1 1 1 2 1 2 1 0 2 6 1 1 1 1 1 0 0 0 1 7 5 2 2 3 1 0 icenv ingWitdhw .8250 .9262 .2962 .9200 .0334 .0300 .9295 .0340 .9260 .0303 .9274 .0333 .2794 .0323 .9215 .0308 .9249 .0309 .9277 .3381 .9292 .9234 .9214 .8293 .1304 .0314 .0332 .7252 .2997 .3273 .3029 .3232 .2985 .3055 .3405 .2598 o r p 6 7 8 2 8 6 4 3 8 2 2 8 9 0 1 4 3 4 8 8 9 2 2 4 2 5 5 5 1 9 5 4 6 9 7 3 36 .00 .00 .00 .00 .00 .00 .00 .00 .00 .00 .00 .00 .00 .00 .00 .00 .00 .00 .00 .00 .00 .00 .01 .00 .00 .00 .00 .00 .00 .00 .01 .00 .00 .00 .00 .00 5 2 0 5 2 4 2 3 8 3 7 2 2 7 7 3 2 2 3 1 2 3 0 8 6 2 6 3 3 5 0 2 3 7 1 5 rfom ingWtlegnh .1787 .1987 .7887 .5998 .1029 .0309 .3088 .5609 .0368 .1909 .4588 .5219 .9668 .5129 .6689 .3091 .9998 .7788 .1129 .5239 .4299 .0669 .0019 .1129 .3159 .9918 .1429 .0328 .2319 .0999 .1959 .2259 .2439 .0359 .1349 .0949 ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± s tiilrrecaeavb itsaecnD D ..816681080± ..813502000± ..190030000± ..815781010± ..914410080± ..915501030± ..814410120± ..916330110± ..818070405± ..915480102± ..914340100± ..024030005± ..814530008± ..919700106± ..186910108± ..195410204± ..198180008± ..148780008± ..169040008± ..271660005± ..250700013± ..169500008± ..169470036± ..158940014± ..280070009± ..159940007± ..189930007± ..016170107± ..519240101± ..519170103± ..619830308± ..719840100± ..619540008± ..919770108± ..319370006± ..219290109± ohpm ec .000± .000± .000± .000± .000± .000± .000± .000± .200± .000± .000± .000± .000± .000± .000± .000± .000± .000± .000± .100± .000± .000± .100± .000± .000± .000± .000± .010± .000± .010± .010± .000± .000± .000± .000± .040± rom tisanD C .5508 .4080 .6308 .3908 .0290 .6380 .7080 .8810 .6840 .5800 .6870 .9840 .2860 .9860 .5890 .7890 .8700 .8660 .8800 .1281 .8610 .8570 .8690 .8340 .9260 .8570 .9480 .8330 .8610 .0261 .8840 .7051 .6810 .1980 .9890 .4930 0 frrrseo1o litaubC B ..05002502± ..00900502± ..22005402± ..00502202± ..00508502± ..00708402± ..00707302± ..00709402± ..10601402± ..00508302± ..00607520± ..00403420± ..00300220± ..00502520± ..00402420± ..00807420± ..00309720± ..00509520± ..00508420± ..00508220± ..00066320± ..00051420± ..02092850± ..00072020± ..00112630± ..00052520± ..00052840± ..00072720± ..00042020± ..00042910± ..00062550± ..00052210± ..00062340± ..00092120± ..00042250± ..00162530± aand fnoy saaym ltaayn tirvn ilirseak lingö lou iirraaybk irend ilazg iirsseh ay traap irsenM iIrzm rsaK tsaaonuKm irsaeyK ililrraekK aonyK isaanM .rsaaKM laugM inpoS isavS razbonT .rfaSU sakU anV tzaogY lanogduZ raaanKm tiranB raahdnA iIrgd k A A A A A B B B D E E kE taH Is . y e n i v o r p e c i h y l o p r o f f o K 7 6 H 8 3 m o r f K 7 s H 7 e m y z M 0 n s G 4 ie P c A . I I s n e o l it b a c a o r i r i k a r i h u n il o i e s a k t lad an n a e N r f f le .oo iev 13 2 3 22 18 15 7 20 3 15 7 12 36 8 9 4 7 16 7 27 12 16 4 6 6 61 2 2 5 91 4 51 6 5 81 4 e h ll N present study. The frequency of the most common allele ranged between 0.637–0.997 in the 18 locations where this enzyme was polymorphic. Mdh65 is the most frequent of the rare alleles in Thrace, whereas Mdh116 is the most frequent rare allele in northeast Turkey. The Mdh65 allele, which is infrequent in honeybees in Africa [ 21 ] and Anatolia ([ 14 ]; and the present study), is common in A. m. ligustica [ 34 ] and A. m. carnica [ 35 ]. The distribution of allele frequencies where the rare alleles are pooled is given in Figure 3. 3.5. Pgi and Me loci Phosphoglucose isomerase (Pgi) and malic enzyme (Me) were invariant in Turkish honeybee populations. The Pgi locus was previously studied in Turkey [ 14 ], and no genetic variability was detected. In this study, the heterozygosities of locations for enzyme loci ranged between 0.012–0.186. Overall average heterozygosity for Turkish honeybees was calculated as 0.072 ± 0.007. This is the highest mean heterozygosity reported in A. mellifera to date. Pamilo et al. [ 27 ] and Sylvester [ 40 ] reported the mean heterozygosity for European honeybees as 0.010 and 0.012, respectively. Later, Sheppard [ 31 ] estimated the mean heterozygosity of A. mellifera from 23 European honeybee colonies as 0.038; he also noted that the mean heterozygosity was rather low in other Apis species, except for A. florea (mean heterozygosities of A. cerana, A. dorsata, A. florea were 0.004, 0.003 and 0.049, respectively). 3.6. Discriminant function analysis Honeybees were allocated to the seven geographic regions of Turkey according to the geographic position of sampling locations. A multiple discriminant function analysis was carried out on the data collected from the samples taken from the seven geographic regions by combining the gene frequencies from electrophoresis and the measurements of morphometric variables. The three axes obtained in the multiple discriminant function analysis explained 87.86% of the total variation. The proportions of variation explained by the first, second, and third axis were 35.54, 32.26, and 20.06% respectively. Two major groups were discriminated by the discriminant function analysis (Fig. 5a). The first group included the honeybees from Thrace. In this group, the main variation was along the first canonical axis. The second group consisted of Anatolian honeybees (honeybees from the Asian part of Turkey), which varied mainly along the second canonical axis. Cubital A, cubital B, Pgm75, Pgm100, Mdh65, Est70, Est100 were the variables with the highest loadings on the first canonical axis, whereas cubital A, cubital B, distance c, Pgm75, Pgm100, Mdh65, Mdh100, Est70, and Est100 were loaded highly on the second canonical axis. In the third canonical axis, wing length, Pgm75, Pgm100, Est70, and Est100 were the variables contributing to the separation of the groups. When Anatolian honeybees alone were subjected to discriminant function analysis on the basis of six geographical regions, distinct clusters of the sampling provinces, representing southeastern Anatolia, the Mediterranean region, central and eastern Anatolia were formed. Samples from the Aegean region could not be distinguished from those in central Anatolia, and samples from the Black Sea overlapped with the those from eastern Anatolia to a large extent (Fig. 5b). Three axes explained 89.68% of the total variation when honeybee populations of Thrace were excluded from the analysis. The variations explained by the three axes were 47.60, 33.80 and 9.28% respectively. Cubital A, cubital B, distance c, Pgm75, Pgm100, Hk110, Mdh65, Mdh100, Est70 and Est100 were the variables that had the highest loadings on the first canonical axis. Cubital A, Pgm75, Pgm100, Hk100, Mdh65 and Mdh100 had high loadings on the second axis. In the third canonical axis, cubital A, distance c, Pgm75, Pgm100, Hk100, Mdh65, Mdh100, Est70 and Est100 were the variables contributing to the separation of the groups. When a phenogram of honeybees from the seven geographic regions was constructed using Mahalanobis distances, the Black Sea and east Anatolian samples clustered very closely. The Aegean and central Anatolia formed a group, and this group together with Mediterranean samples made up a larger cluster. Thrace and southern Anatolian samples remained as distinct units within this phenogram (Fig. 6). When multiple regression analysis was applied to the morphometric and electrophoretic variables using latitude and longitude as independent variables, eight out of 10 morphometric variables turned out to be significantly dependent on latitude (cubital B, distance c, distance d, wing length, wing width, metatarsus width, femur and tibia length). Cubital B was also significantly dependent on longitude. Among the 12 gene frequencies, six of them showed a significant relationship with latitude (Pgm75, Pgm100, Hk110, Mdh65, Mdh100, and Est70). Mdh65 and Mdh100 were also significantly dependent on longitude (Tab. III). 4. DISCUSSION Honeybees in Turkey show a high level of morphometric variation. Of the ten characters studied, five (distance c, distance d, wing length, wing width and metatarsus width) were found to be significantly different between localities (P < 0.05). The significant regressions of morphometric and electrophoretic variables on latitude and longitude display a structured pattern in the distribution of populations. The spatial nature of this pattern is most likely the result of evolutionary forces acting on the honeybee populations. This hypothesis was supported by a spatial autocorrelation analysis conducted to further determine relationships among honeybee populations of Anatolia [ 15 ]. Morphometric variables that showed significant regressions on latitude also had high loadings on the first axis in the principal component analysis [ 15 ]. This axis, known as the size axis [ 13 ], allows us to conclude that the size of the honeybees increases with increasing latitude. Daly et al. [8] showed similar clinal geographic variation in morphometric characters in feral colonies of California. Based on the UPGMA phenogram, the differentiation in honeybees in Turkey has been maintained, despite extensive migratory beekeeping. According to Ruttner [ 30 ], A. m. anatoliaca is distributed throughout central Anatolia, the Aegean, the Mediterranean, and a large part of the Black Sea region. A. m. meda is distributed in southeastern Anatolia, A. m. caucasica in northeastern Anatolia, and A. m. carnica in Thrace. This assessment of subspecies distribution is largely supported by electrophoretic data and our morphometric assessments on a reduced number of characters in honeybee populations. However, the small set of samples from southeastern Anatolia form a distinct cluster that appears to belong to A. m. syriaca, based on values of CI, wing length, and body size [ 11 ]. Similarly, honeybee populations in the Mediterranean region, isolated from the rest of the Anatolian population by the Taurus mountain range, appear to form another distinct cluster. Further studies are needed to determine the taxonomic status of these honeybee populations by including additional samples and a full morphometric analysis. The honeybees of Turkey were separated into two groups, the European and the Anatolian, by discriminant function analysis. Kirklareli and Edirne honeybee populations (in Thrace) had the highest Mdh65 gene frequencies, with the highest loadings on the first canonical axis. We observed that the Kirklareli honeybee population also had the highest tibia length. Anatolian honeybees were separated along the second canonical axis, with distance c, tibia length, Pgm75 and Pgm100 variables with high loadings. The Black Sea and east Anatolian samples had the highest, whereas the Mediterranean samples had the lowest distance c values, which separated these two groups on the second axis. Of the 36 provinces from which the samples were taken, only one province (K. Maras) was fixed for the Pgm75 allele based on a very small sample size (12 worker bees). In all other provinces, Pgm showed a high degree of polymorphism. Besides Pgm100 and Pgm75, two additional rare alleles (Pgm45 and Pgm63) were observed. There were strong deviations from Hardy–Weinberg equilibrium in a number of provinces for Pgm (P < 0.001). Hatay and S. Urfa samples showed the most extreme deviations. In Hatay, 269 out of 271 individual honeybees were heterozygous for Pgm75/100; in S. Urfa, all samples (14 individuals) were heterozygous for the same alleles. Bingöl, Kirklareli, and Elazig also showed some degree of deviations in favor of heterozygotes. Kandemir and Kence [ 14 ] found four alleles (Hk87, Hk100, Hk110, and Hk120) in central Anatolian honeybee populations; and in the present study we found an additional allele, Hk77, which has not been reported previously. Out of seven alleles at the Mdh locus (Mdh55, Mdh65, Mdh80, Mdh87, Mdh100, Mdh116 and Mdh133), reported by various authors in different honeybee populations [ 1–3, 5, 6, 12, 16, 17, 21, 25, 26, 32–35 ], five of them (Mdh65, Mdh87, Mdh100, Mdh116, and Mdh133) have been observed in honeybee populations of Turkey [14]. The frequency of the Mdh65 allele in Turkey has been found to be highly reduced; and in southern and southeastern Anatolia, the Mdh locus has become invariant (Mdh100). This relationship has also been seen in the significant linear regression of Mdh65 and Mdh100 on latitude and longitude. This type of clinal variation has been reported by Nielsen et al. [ 23 ] in Mdh allozymes in Europe, California, and Brazil with the suggestion that selection may be involved in many clines. There is some evidence that fitness differing in Mdh genotypes may occur, as recent studies have shown differences in temperature optima [7] and differential oxygen consumption during hovering [4]. One important result regarding the electrophoretic analysis is the observation of a large number of rare alleles (Tab. II). The existence of rare alleles in a population suggests that there has not been a recent bottleneck for the population. The observations of rare alleles, the presence of four subspecies and the detection of high genetic diversity as reflected in the high heterozygosity support the argument that Anatolia has been a genetic center for honeybee populations in the Near East. This study is the most extensive survey yet made of the electrophoretic and morphometric variation in honeybee populations in the Near East. However, extended studies including additional morphometric characters and samples from surrounding countries (Syria, Iraq, Iran, Georgia and Armenia) would certainly need to include a more complete picture of the genetic variability in the Middle East and Asia. Résumé – Variation génétique et morphométrique des populations d’abeilles domestiques (Apis mellifera L.) en Turquie. Selon l’analyse statistique multivariée des données morphométriques, l’Asie mineure semble être un centre de diversification génétique pour les races d’abeilles domestiques qui peuplent cette région, mais peu d’études ont été consacrées à la diversité génétique et morphométrique des abeilles d’Anatolie. Six systèmes enzymatiques ont été étudiés pour déterminer la variabilité génétique des populations d’abeilles en Turquie : enzyme malique (Me), phosphoglucomutase (Pgm), estérase-3 (Est), hexokinase (Hk), phosphoglucose isomérase (Pgi), malate déshydrogénase (Mdh). Dix caractères morphométriques ont été mesurés pour déterminer l’étendue de la variation morphométrique : huit caractères selon Ruttner [ 30 ] (indice cubital A et B, longueur et largeur de l’aile antérieure, longueur du fémur, longueur du tibia, longueur et largeur du métatarse) et deux caractères selon Nazzi [ 20 ] (distances c et d de l’aile antérieure) (Tabs. I et II). La majorité des variables morphométriques et enzymatiques ont des relations linéaires significatives quand on fait une régression sur la latitude et la longitude (Tab. III). On a trouvé que quatre des six systèmes enzymatiques étaient polymorphes et présentaient 16 isozymes. L’hétérozygosité moyenne était de 0,072 – 0,007. Les deux types de données, morphométriques et électrophorétiques, ont été utilisées pour discriminer les populations d’abeilles turques. Les abeilles européennes et les abeilles d’Anatolie sont discriminées par le 1er axe canonique, et les abeilles d’Anatolie se séparent le long du 2e axe. Les variables morphométriques ont été aussi efficaces que les variables électrophorétiques pour discriminer les populations d’abeilles. Un résultat important concernant l’analyse électrophorétique est la présence d’un grand nombre allèles rares (Figs. 1 à 4). L’existence d’allèles rares dans une population suggère qu’il n’y a pas eu de goulot d’étranglement récent. L’observation d’allèles rares, la présence de quatre sous-espèces connues et la mise en évidence d’une diversité génétique élevée démontrée par la forte hétérozygosité confirment l’argument selon lequel l’Anatolie a été un centre de diversification génétique pour les populations d’abeilles domestiques au Proche-Orient. Apis mellifera anatoliaca / A. m. caucasica / A. m. meda / A. m. syriaca / génétique population / variabilité génétique / morphométrie / électrophorèse / Turquie Zusammenfassung – Genetische und morphometrische Variation in türkischen Honigbienenpopulationen (Apis melli fera L.). Zur Bestimmung der genetischen Variabilität der Honigbienenpopulation in der Türkei wurden sechs Enzymsysteme (Malat-Enzym, Phosphoglucomutase, Esterase-3, Hexokinase, Phosphoglucoseisomerase, Malatdehydrogenase) untersucht. Zur Bestimmung morphologischer Variationen wurden zehn morphometrische Charaktere (Cubitalader A und B, Abstand c und d, Länge und Breite des Flügels und des Metatarsus, Länge des Femur und der Tibia) vermessen. Die meisten morphometrischen und elektrophoretischen Variablen zeigten signifikante lineare Beziehungen in der Regression auf die geographische Breite und Länge (Cubital B, Abstand c und d, Flügellänge und -breite, Breite des Metatarsus, Länge von Femur und Tibia, Pgm75, Pgm100, Hk110, Mdh65, Mdh100 und Est70). Bei vier von sechs Enzymsystemen wurden Polymorphismen mit insgesamt 16 Isoenzymen gefunden. Die mittlere Heterozygosität wurde zu 0,072 – 0,007 berechnet. Zur Unterteilung der türkischen Bienen wurden sowohl die morphometrischen als auch die elektrophoretischen Daten verwendet. Europäische und anatolische Honigbienen waren auf der ersten canonischen Achse unterschiedlich, die anatolischen Bienen spalteten sich entlang der zweiten Achse weiter auf. Morphometrische und elektrophoretische Variablen waren gleich gut zur Unterscheidung der Honigbienenpopulationen geeignet. Ein wichtiges Resultat der elektrophoretischen Analyse war die grob e Zahl seltener Allele. Dieser Befund legt nahe, dass in der jüngeren Populationsentwicklung kein Flaschenhals aufgetreten ist. Der Befund seltener Allele, die Anwesenheit von vier bekannten Subspezies, sowie die durch die hohe Heterozygosität angezeigte grob e genetische Diversität stützen die Auffassung, dass Anatolien ein Entwicklungszentrum der Honigbienenpopulationen des Nahen Ostens dargestellt hat. Apis mellifera anatoliaca / A. m. caucasica / A. m. meda / A. m. syriaca / Populationengenetik / genetische Variabilität / Morphometrie / Elektrophorese / Türkei ACKNOWLEDGMENTS The authors wish to thank to all who assisted in the collection, preparation, and measurement of the samples. Thanks are due to the Ministry of Agriculture and Rural Affairs of Turkey for providing assistance during the collection of samples, and to the beekeepers. This work was supported by grant VHAG-1077 from the Turkish Scientific and Technical Research Council to Meral Kence. [1] [2] [3] [4] [5] [6] [7] [8] Badino G. , Celebrano G. , Manino A. , Population structure and Mdh-1 locus variation in Apis mellifera ligustica , J. Hered . 74 ( 1983 ) 443 - 446 . Badino G. , Celebrano G. , Manino A. , Longo S. , Enzyme polymorphism in the Sicilian honeybee , Experientia 41 ( 1985 ) 752 - 754 . 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Irfan Kandemir, Meral Kence, Aykut Kence. Genetic and morphometric variation in honeybee (Apis mellifera L.) populations of Turkey, Apidologie, 343-356, DOI: doi:10.1051/apido:2000126