As a part of breeding project, six Apis mellifera inbreed lines (named line A, C, D, E, F and G) were breed and cultivated for several years via an artificial insemination method. In this study, I evaluated the six inbreed lines according to morphological values and hygienic behavior. The morphometr...
As a part of breeding project, six Apis mellifera inbreed lines (named line A, C, D, E, F and G) were breed and cultivated for several years via an artificial insemination method. In this study, I evaluated the six inbreed lines according to morphological values and hygienic behavior. The morphometric characters measured using basitarsus length (ML), proboscis length (PL), fore wing length (FWL), cubital vein, distance (a), cubital vein, distance (b) and counted the number of hook (Ha). The result of that measured means of each body part showed significant differences by one-way ANOVA and post hoc Tukey HSD test (p90%), two inbreed lines exhibited intermediate behavior and one inbreed line was non-hygienic (removal of dead brood <60%). Among them, the removal rate in 24 h showed no significant differences by one-way ANOVA test (p<0.05). Microsatellites, a special class of repetitive DNA sequences, have become one of the most popular markers of genetic polymorphism, especially as next-generation sequencing (NGS) methods have become available to streamline the identification of such markers. Based on 45,476 contigs generated by NGS, I obtained 205 Mbp of nucleotide sequences from six inbreed lines of Apis mellifera. In total, 20,580 repeat motifs were identified. Di-nucleotide repeats, comprised mostly of [AT]n motifs, were two-fold more common than tri-nucleotide repeats. I arbitrarily selected 50 microsatellite loci with high repeat numbers and were covered by overlapping contigs. PCR amplification yielded clean fragments from 34 of these loci and nine were selected to differentiate the inbreed lines. Genotyping at these nine loci revealed 4 ~ 14 alleles per locus, with average 10.11, as well as 0.1875 ~ 0.9592 per-locus observed heterozygosity and 0.5432 ~ 0.8669 per-locus polymorphic information content. Thus, some of these loci are profoundly variable. Phylogenetic analysis using the nine microsatellite loci separated individual honeybees from different inbreed lines very well, suggesting that these markers maybe used to trace the six inbreed lines of A. mellifera maintained in Korea. To select for honeybees with high honey production, I established various cross line from six inbreed lines. From those cross lines, I eventually selected AC♀ × D♂ hybrid line with high yield of honey per hive, which was 28.6% higher than that in the control. To evaluate selected AC♀ × D♂ hybrid line, I investigated whether high efficiency in honey collection of AC♀ × D♂ hybrid line is associated with population development, disease resistance, hygienic behaviour and defensive behaviour. First, I measured brood development using population size of eggs, broods and adult bees. The result showed that the population of adult bees expands more in the AC♀ × D♂ hybrid line despite the lower eggs and brood population. There was a positive correlation between the number of adult bees and honey yield. The hygiene behaviour was assessed using the freeze-killed method to validate disease resistance in the colony. In our experiment, the removal rate of AC♀ × D♂ hybrid lines was 74% and 117% higher compared with F1 colony and control, which was significantly higher than those observed in other lines. The defensivity test was conducted according to measured a number of sting when defensive responses. In the present study, the level of defensive behaviour in the AC♀ × D♂ hybrid line (0.33–0%) was similarly to that found in low-defensive colonies (0.67–2.0%).
As a part of breeding project, six Apis mellifera inbreed lines (named line A, C, D, E, F and G) were breed and cultivated for several years via an artificial insemination method. In this study, I evaluated the six inbreed lines according to morphological values and hygienic behavior. The morphometric characters measured using basitarsus length (ML), proboscis length (PL), fore wing length (FWL), cubital vein, distance (a), cubital vein, distance (b) and counted the number of hook (Ha). The result of that measured means of each body part showed significant differences by one-way ANOVA and post hoc Tukey HSD test (p90%), two inbreed lines exhibited intermediate behavior and one inbreed line was non-hygienic (removal of dead brood <60%). Among them, the removal rate in 24 h showed no significant differences by one-way ANOVA test (p<0.05). Microsatellites, a special class of repetitive DNA sequences, have become one of the most popular markers of genetic polymorphism, especially as next-generation sequencing (NGS) methods have become available to streamline the identification of such markers. Based on 45,476 contigs generated by NGS, I obtained 205 Mbp of nucleotide sequences from six inbreed lines of Apis mellifera. In total, 20,580 repeat motifs were identified. Di-nucleotide repeats, comprised mostly of [AT]n motifs, were two-fold more common than tri-nucleotide repeats. I arbitrarily selected 50 microsatellite loci with high repeat numbers and were covered by overlapping contigs. PCR amplification yielded clean fragments from 34 of these loci and nine were selected to differentiate the inbreed lines. Genotyping at these nine loci revealed 4 ~ 14 alleles per locus, with average 10.11, as well as 0.1875 ~ 0.9592 per-locus observed heterozygosity and 0.5432 ~ 0.8669 per-locus polymorphic information content. Thus, some of these loci are profoundly variable. Phylogenetic analysis using the nine microsatellite loci separated individual honeybees from different inbreed lines very well, suggesting that these markers maybe used to trace the six inbreed lines of A. mellifera maintained in Korea. To select for honeybees with high honey production, I established various cross line from six inbreed lines. From those cross lines, I eventually selected AC♀ × D♂ hybrid line with high yield of honey per hive, which was 28.6% higher than that in the control. To evaluate selected AC♀ × D♂ hybrid line, I investigated whether high efficiency in honey collection of AC♀ × D♂ hybrid line is associated with population development, disease resistance, hygienic behaviour and defensive behaviour. First, I measured brood development using population size of eggs, broods and adult bees. The result showed that the population of adult bees expands more in the AC♀ × D♂ hybrid line despite the lower eggs and brood population. There was a positive correlation between the number of adult bees and honey yield. The hygiene behaviour was assessed using the freeze-killed method to validate disease resistance in the colony. In our experiment, the removal rate of AC♀ × D♂ hybrid lines was 74% and 117% higher compared with F1 colony and control, which was significantly higher than those observed in other lines. The defensivity test was conducted according to measured a number of sting when defensive responses. In the present study, the level of defensive behaviour in the AC♀ × D♂ hybrid line (0.33–0%) was similarly to that found in low-defensive colonies (0.67–2.0%).
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