• International Journal of Industrial Entomology and Biomaterials
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• v.27 no.2
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• pp.271-276
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• 2013

The Asian honeybee, Apis cerana, is a native honeybee species in Korea which is important in agriculture for pollination and honey production. For better understanding of the physiology of A. cerana, high-throughput Illumina transcriptome sequencing was performed to analyze the gene expression profiles of queen, worker, and larva. A total of 219,799,682 clean reads corresponding to 22.2 Gb of nucleotide sequences was obtained from the whole body total RNA samples. The Apis mellifera reference mRNA sequence database was used to measure the gene expression level with Bowtie2 and eXpress software, and the Illumina short reads were then mapped to 11,459 out of 11,736 A. mellifera reference genes. Total of 9,221 genes with FPKM value greater than 5 of each sample group were subjected to eggNOG with BLASTX for gene ontology analysis. The differential gene expression between queen and worker, and worker and larva were analyzed to screen the overexpressed genes in each sample group. In the queen and worker sample group, total of 1,766 genes were differentially expressed with 887 and 879 genes overexpressed over two folds in queen and worker, respectively. In the worker and larva sample group, total of 1,410 genes were differentially expressed with 1,009 and 401 genes overexpressed over two folds in worker and larva, respectively.

• BMB Reports
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• v.36 no.6
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• pp.572-579
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• 2003

An expressed sequence tag (EST) library was established from the hypopharyngeal glands of Apis cerana. Sixty-six recombinant clones, possessing inserts >500 bp, were randomly selected and unidirectional sequenced. Forty-two of these (63.6%) were identified as homologues of Major Royal Jelly Proteins families 1, 2, 3, and 4 of A. mellifera (AmMRJP) for which MRJP1 was the most abundant family. The open-reading frame of the MRJP1 homologue (AcMRJP1) was 1299 nucleotides that encoded 433 deduced amino acids with three predicted N-linked glycosylation sites. The AcMRJP1 sequence showed 93% and 90% homologies with nucleotide and deduced amino acid sequences of AmMRJP1, respectively. Two complete transcripts of apisimin, and one and two partial transcripts of $\alpha$-glucosidase and glucose oxidase, were also isolated. In addition, the royal jelly proteins of A. cerana were purified and characterized using Q-Sepharose and Sephadex G-200 column chromatography. The native forms of protein peaks A1, A2, B1, and C1 were 115, 55, 50, and 300 kDa, respectively. SDS-PAGE analysis indicated that A1 and C1 were dimeric and oligomeric forms of the 80 kDa and 50 kDa subunits, respectively. The ratio of the total protein quantities of A1 : A2 : B1 : C1 were 2.52 : 4.72 : 1 : 12.21. Further characterization of each protein, using N-terminal and internal peptide sequencing, revealed that the respective proteins were homologues of MRJP3, MRJP2, MRJP1, and MRJP1 of A. mellifera.

• Journal of Apiculture
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• v.35 no.1
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• pp.9-19
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• 2020

The questionnaire survey for Apis cerana beekeepers and professionals on improved native bee hives was carried out and we compared the temperature, relative humidity(RH), and weight changes of 4 improved hives(Chungju, Miryang, Hanam, and Suwon) from May 1, 2019 to January 31, 2020. Beekeepers need vertical feeder, hive stand, entrance block, and separating panel as hive accessory devices. The average temperatures within brood area were kept constantly (31.3~35.1℃) and the low daily variances of temperature (≤1℃) in Chungju hive among tested hives were observed. The daily temperature variances in the separated space and on the top of winter cluster were not different among 4 hives. In correlation between the temperature of brood area and the number of combs, Chungju hive showed the highest correlation(80.4%) and between the temperature on top of winter clusters and outside temperature, 4 hives showed high positive correlation(76.8~87.1%). RH of brood area(45~60%) in all hives were kept relatively low and constant compared to the outside RH(60~85%). The stablest RH on the top of winter cluster was observed in Suwon hives (65~75%) The highest cumulative weight increase among hives and the high positive correlation(65~67%) between the change of cumulative hive weight and combs number of hives were shown in the Miryang and Chungju. Based on these results, A. cerana bees are able to manage constant temperature and RH within hives area for their colony life, which also effected by the types of hive.

• International Journal of Industrial Entomology and Biomaterials
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• v.18 no.2
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• pp.139-142
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• 2009

The histological and transmission electron microscopic studies revealed the synthesis activity predominantly in the hypopharyngeal glands of the nurse bees. The biochemical analysis of both, the hypopharyngeal gland extract and royal jelly elucidated unequivocally the proteins and lipids as the major constituents. Further the SDS-PAGE of hypopharyngeal gland extract showed about 17 protein bands, perhaps 14.10, 20.00, 29.00 and 43.00 kDa predominantly while that of royal jelly revealed only two protein bands of 29.00 and 43.00 kDa molecular weight suggesting them as the major royal jelly proteins (MRJP). The lipid profile of royal jelly consists of triglycerides, cholesterol, HDL, LDL and VLDL.

• Proceedings of the Korean Society of Applied Entomolohy Conference
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• 2005.05a
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• pp.126-126
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• 2005

• BMB Reports
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• v.38 no.1
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• pp.49-57
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• 2005

Major Royal Jelly Protein cDNAs of Apis cerana (AcMRJP) were cloned and characterized. The open reading frames (ORFs) of the AcMRJP1 and AcMRJP2 genes were 1302 and 1392 nucleotides, encoding 433 and 463 amino acid residues, respectively. The sequence divergences between AcMRJP1 and AcMRJP2 and their corresponding protein families in A. mellifera were 0.0618 and 0.0934 at the nucleotide level and 0.0912 and 0.1438 at the protein level, respectively. Phylogenetic analysis supports the orthologous similarity between these proteins. The deduced amino acids indicated high essential amino acid contents of AcMRJP1 and AcMRJP2 (47.5 and 44.8%, respectively). The genomic organization of both AcMRJP1 and AcMRJP2 was determined. Both the AcMRJP1 (3663 bp) and AcMRJP2 (3963 bp) genes contained six exons and five introns, where all boundaries conformed to the GT/AG rule. AcMRJP1 and AcMRJP2 cDNAs were cloned into pET17b, and both the recombinant (r) AcMRJP1 (47.9 kDa) and rAcMRJP2 (51.7 kDa) were expressed in the insoluble form. Western blot analysis and N-terminal sequencing of the solubilized proteins revealed successful expression of rAcMRJP1 and rAcMRJP2 in vitro. The yields of the purified rAcMRJP1 and rAcMRJP2 were approximately 20 and 8mg protein per liter of the flask culture, respectively.

• International Journal of Industrial Entomology and Biomaterials
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• v.31 no.2
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• pp.62-69
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• 2015

The Asian cavity-nesting honeybee, Apis cerana (Hymenoptera: Apidae), has been extensively studied for its biogeography and genetic diversity, but the molecules utilized in past studies were mainly ~90 bp long mitochondrial non-coding sequences, located between $tRNA^{Leu}$ and COII. Thus, additional molecular markers may enrich our understanding of the biogeography and genetic diversity of this valuable bee species. In this study, we reviewed the public genome database to find introns of cDNA sequences, with the assumption that these introns may have less evolutionary constraints. The six introns selected were subjected to preliminary tests. Thereafter, two introns, titled White gene and MRJP9 gene, were selected. Sequencing of 552 clones from 184 individual bees showed a total of 222 and 141 sequence types in the White gene and MRJP9 gene introns, respectively. The sequence divergence ranged from 0.6% to 7.9% and from 0.26% to 17.6% in the White gene and the MRJP9 introns, respectively, indicating higher sequence divergence in both introns. Analysis of population genetic diversity for 16 populations originating from Korea, China, Vietnam, and Thailand shows that nucleotide diversity (π) ranges from 0.003117 to 0.025837 and from 0.016541 to 0.052468 in the White gene and MRJP9 introns, respectively. The highest π was found in a Vietnamese population for both intron sequences, whereas the nine Korean populations showed moderate to low sequence divergence. Considering the variability and diversity, these intron sequences can be useful as non-mitochondrial DNA-based molecular markers for future studies of population genetics.

• Journal of Apiculture
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• v.33 no.4
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• pp.261-268
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• 2018

Queen honey bees (Apis cerana) can be reared artificially on demand by the use of grafting technique. The technique consists of grafting young worker larvae into queen cell cups and raising in a queenless strong colony. As the age of grafted larvae for queen rearing exhibited several aspects related to quality and reproductive potential of queen, this study were conducted to investigate the influence of age of grafted larva on morphological characteristics and lifespan of queen, and the growth of colony she headed. Our results demonstrated that queens reared from young worker larvae (i.e., less than 1-day old larvae) were significantly larger in size (i.e., body weight and thorax width) than that of queens reared from 2-day old worker larvae. Moreover, queens reared from younger worker larvae initiated egg-laying earlier, stored more spermatozoa in spermatheca and had a longer lifespan compared to queens raised from older worker larvae. We also found a significant positive effect of queen grafting age on the production of worker and drone brood, adult worker population in colonies headed by queens reared from younger larvae. These findings suggested that rearing queens from brood grafted at the earliest possible age could increase the reproductive potential of queen as well as fitness of colony she head.

• Journal of Apiculture
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• v.33 no.4
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• pp.251-260
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• 2018

Instrumental insemination of honey bee is an attractive alternative to natural mating in breeding program as it allows mating crosses between desirable queen and specific drone. However, nursery condition that the queen is kept before and after insemination is major factor affected to the performance of instrumentally inseminated queen. In this study, we evaluated the influences of three different nursery-conditions of push-in cages, mini nuclei and normal colonies on number of spermatozoa stored in the spermatheca, body weight, onset of ovipositon and performance of instrumentally inseminated Apis cerana queen. Our results demonstrated that instrumentally inseminated queens kept in mini nuclei and in normal hives showed no significant difference in queen's weight (159.8 and 166.2mg, respectively), number of spermatozoa in spermatheca ($2.02{\times}10^6$ and $2.76{\times}10^6$, respectively), proportion of queen supersedure (33.3 and 66.7% queen survival at 11 months after oviposition, respectively) and brood production, compared to naturally mated queens. In contrast, instrumentally inseminated queens kept in push-in cages showed significant difference of those above data in comparison to queens mated naturally. Our results suggested that instrumentally inseminated queens could be kept in mini nuclei containing about 1.000 attendant bees to have desirable performance of queen whereas the push-in method should be practiced for the purpose of using queen in the length of time less than 7 months.

• Korean Journal of Environmental Agriculture
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• v.8 no.1
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• pp.47-54
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• 1989

This study was conducted to investigate insecticide toxicities to a honeybee, Apis cerana F. being raised in Korea and its detoxifying enzyme activities. In order to determine the appropriate usage of insecticides, median effective dose and detoxifying enzyme activities to seven insecticides were observed. Various detoxifying enzymes, including microsomal oxidases, glutathione S-transferases, esterases, and DDT-dehydrochlorinase were assayed in the midguts of adult worker bees as the enzyme source. Of the insecticides used, $LC_{50}$ value in DDT treatment was the highest as 19ppm, and that in EPN treatment was the lowest as 0.75ppm. Sublethal exposures of honeybees to various insecticides had some effects on microsomal enzyme activities. Aldrin epoxidase activity was inhibited by malathion and demeton S-methyl treatment. N-demethylase activity was induced by carbaryl treatment. Of the glutathione S-transferases, aryltransferase(DCNB conjugation) activity was significantly induced by diazinon, and moderately induced by malathion. Of the esterases, ${\alpha}-NA$ esterase activity was moderately inhibited by malathion and permethrin. Carboxylesterase and acetylcholinesterase activity were not affected by the sublethal exposure of honeybee to the insecticides. Sublethal exposure of honeybee to the insecticides had no effect on DDT- dehydrochlorinase activity, except carbaryl, malathion and demeton S-methyl were inhibited.