Master thesis

 

Stevanovic Jevrosima

 

Investigations of morphometric and chromosomal variability in diversity preserving of Carniolan honey bee (Apis mellifera carnica Pollmann, 1879) in Serbia

 

 

SUMMARY

 

The concept of preservation of biological diversity takes into consideration all aspects of living nature on Earth, the entire biological resources of genes, species and ecosystems, as well as the mankind necessity for further prosperity and survival. At the first plase, within this concept are simple and direct verification and systematization of the entire biodiversity as a foundation stone of all present and future human activities.

The aim of this study was to investigate morphometric and chromosomal diversities of indigenous populations of the carniolan honeybee A. m. carnica in order to preserve the genetic diversity of this subspecies. The established aim is achieved by realizing the following tasks: 1. Microecological characterization of regions where honeybees of the Banat, Timok and Syenichko–Peshterski ecotypes were sampled; 2. Morphometric analysis of honeybees of the sampled ecotypes; 3. Chromosomal analysis of the honeybees of the mentioned ecotypes.

For microecological analysis, all climatic data the for three-year research period (1997-1999) were taken over from the Federal Weather Buerau and were used for designing climadiagrams. Orographic data were gained from the topographic maps (1 : 50 000) of the investigated regions, available from the Military Geographic Institute. The floristic data, for the regions where sampling was done, were obtained by monitoring the visited localities and comparing the obtained results with the relevant literature data.

For morphometric analyses, altogether 900 samples were collected from 30 different localities (10 localities from each investigated region). 30 adult workers were sampled from each locality and their fore wings, hind wings and hind legs were prepared. The basic parameters of descriptive statistics were calculated for all 30 analysed morphometric characters (16 lengths and 14 venation angles) – for the whole specimen, separately, for each ecotype and locality. In order to ascertain the significance of individual differences between morphometric characters (between ecotypes and between localities within each ecotype) the analysis of variance (ANOVA) was performed. The canonical discriminant analysis (CDA) was applied to morphometric data in order to evaluate level of divergency and to identify elements of morphometric discrimination between the ecotypes.

For chromosomal analyses of honeybees, 12 worker prepupae from each locality were sampled. Since prepupae were collected from 10 localities of each investigated region (inhabited with Banat, Timok and Syenichko–Peshterski honeybee ecotypes), it means that 120 prepupae of each ecotype were collected. Only the cerebral ganglia from living worker prepupae, nine to eleven days old, with star-like, redish eyes, were used for the cytogenetic analysis. Chromosomes from the cerebral ganglia were prepared in accordance with the procedure of Hoshiba et al. (1995), adapted to the requirements of this study. G – banding of chromosomes was done by the tripsin-method of Ronne (1991), which is a modification and an improvement of those established by Seabright (1971). The Analysis of distribution of euchromatin and heterochromatin regions were performed on honeybee chromosomes of Banat, Timok and Syenichko–Peshterski ecotypes in order to determine the interecotype chromosomal differences between honeybees in Serbia.

The results of microecological investigations show orographic, as well as climatic and floristic differences between the regions inhabited with the investigated honeybee ecotypes. The results of the descriptive statistics and analysis of variance of morphometric characters confirmed the existence of inter- and intraecotype honeybee variabilities in Serbia. The greatest interecotype differences were recorded between Syenichko–Peshterski and Banat honeybee ecotypes; less between Timok and Banat and the least differences were noticed between honeybeees of Syenichko–Peshterski and Timok ecotypes. The greatest intraecotype variability was registered inside Syenichko–Peshterski ecotype, less within Timok ecotype and the least within Banat ecotype. The analysis of variance (ANOVA) indicated that intraecotype variability is minor in relation to the interecotype variability.

The Canonical Discriminant Analysis CDA of all the analysed morphometric characteristics revealed 100% of the total variation in the first two canonical axes. The values of standardized coefficients indicated that in total specimen the greatest proportion of the variance on the first canonical axis was expressed by differences in value of angles G18, E9 and B4. The greatest contributions to variance on the second axis were made by differences in fore wing length (FWLN), femur length (FELN) and in value of angle G18. Two-dimensional plots of scores of the first two canonical variables discriminated three groups (ecotypes) from each other: Banat and Syenichko–Peshterski ecotypes were discriminated on the first canonical axis due to the differences in values of angles G18, E9 and B4, while Timok ecotype was separated from Banat and Syenichko–Peshterski ecotypes on the second axis due to the differences in fore wing length (FWLN), femur length (FELN) and in value of angle G18.

Chromosomal analyses revealed differences between Timok and Banat honeybee ecotypes in case of chromosomes 2, 4, 11 and 13. Between Timok and Syenichko–Peshterski ecotypes, differences in chromatic region redistribution were observed for chromosomes 1, 2, 15 and 16. The greatest differences in G-band distribution were registered between Syenichko – Peshterski and Banat ecotypes in case of chromosomes 1, 2, 4, 11, 12, 13, 15 and 16.

The variability in phenotype and genotype of the carniolan honeybee, confirmed after morphometric and chromosomal analyses is the result of various environmental factors, due to the fact that samplings were done in regions some 200 km apart, with different climatic, geographic and floristic features. All the results point to a great inter- and intraecotype variability of the carniolan honeybee in Serbia. The analysed ecotypes A. m. carnica represent a remarkable genetic resource for a highly selectioned breed impovement. Therefore, it is necessary to undertake protection measures for these ecotypes by forming segregated apiaries in their indigenous regions.