Transcriptional profiling reveals the molecular bases of immune regulation in Apis mellifera in response to Ascosphaera apis infection

Bo Xu,Yingzhe WANG,Shixin ZHU,Haizhu ZHOU,Changlong GOU,Wenlong DONG,Yu Wang,Yunhang GAO,Hongxia MA 한국곤충학회 2019 Entomological Research Vol.49 No.1

Chalkbrood, which results from Ascosphaera apis infection, is one of the bee diseases that causes serious damage to the bee colony. Understanding the molecular bases underlying immune response to chalkbrood disease would facilitate the genetic breeding of bees by selecting races with superior chalkbrood resistance. In this study, transcriptome sequencing was performed to identify genes and pathways involved in the immune response to As. apis infection in A. mellifera larvae. In total, 2,890 differentially expressed genes (DEGs) (FDR < 0.001) were identified between the healthy and As. apis infected bee larvae, including 2,214 up‐regulated and 676 down‐regulated unigenes. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway classification of the DEGs revealed association with development, energy metabolism, signal transduction, disease, and immune response. Among the immune‐related genes, p38, toll‐like receptors (TLRs), integrin, and antimicrobial peptides were up‐regulated under As. apis infection. This study provides valuable gene transcriptional information towards the investigation of molecular mechanisms related to chalkbrood immune response and host pathogenesis in A. mellifera.

Estimation of C-derived introgression into A. m. mellifera colonies in the Russian Urals using microsatellite genotyping

Rustem A. Ilyasov,Myeong‑Lyeol Lee,Ural Yunusbaev,Alexey Nikolenko,Hyung‑Wook Kwon 한국유전학회 2020 Genes & Genomics Vol.42 No.9

Background Marker-assisted selection is well established in animal breeding method of selecting individuals with desirable traits in a breeding scheme based on DNA molecular marker patterns. Objective Genetic diversity and C-derived admixture into local purebred gene pool of A. m. mellifera colonies was assessed using polymorphism of nine microsatellite loci in order to provide further marker-assisted selection of desired honey bee colonies. Methods The genetic diversity and the level of C-derived introgression into A. m. mellifera colonies in the Shulgan-Tash Nature Reserve (Russia) was assessed based on nine microsatellite loci (ap243, 4a110, A24, A8, A43, A113, A88, Ap049, A28), which were analized using the fragment analysis of the PCR products in Applied Biosystems 3130 DNA Analyzer. Phylogenetic relationship of colonies was evaluated using Neighbor-Joining methods with Cavalli-Sforza and Edwards genetic distance using the PHYLIP 3.68. The model-based Bayesian clustering algorithm implemented in STRUCTURE 2.3.3 was employed to infer membership and introgression proportions (Q-value). Results In the Shulgan-Tash Nature Reserve colonies of A. m. mellifera subdivided into four groups by level of C-derived introgression. Only fve colonies of A. m. mellifera had C-derived introgression which varied from 0.5 to 2%. The genetic diversity in colonies of A. m. mellifera varied from 0.12 to 0.40. The Neighbor-Joining tree demonstrates the genetic relationship of A. m. mellifera colonies, which subdivided into three groups with diferent levels of C-derived introgression. Group 1 combined fve honey bee colonies Bort_1, Bort_2, Bort_3, Baisalyan_1, and Kush_7 with a fraction of introgression close to 0.000 and genetic diversity from 0.20 to 0.25. Conclusion The results showed the excellence of nine microsatellite loci genotyping in estimation of genetic diversity, distinguishing the two European evolutionary lineages M and C and estimating C-derived introgression. These genetic parameters can be applied further to perform the marker-assisted selection of purebred dark European honey bees.

Differential hygienic behavior of Apis cerana F. and Apis mellifera L. to Sacbrood virus infection

Choi Yong Soo,Park-HeeGeun,Frunze Olga 한국응용곤충학회 2022 Journal of Asia-Pacific Entomology Vol.25 No.4

Beekeeping with Apis cerana of Korean apiculture is facing with serious colony collapse caused by invasive Sacbrood virus (SBV) disease. This fatal brood disease was the main reason of more than 90% colony lost in Korea leading almost the extinct crisis. Sacbrood virus can infect either larvae or adult honeybees, with a higher sensibility of larvae to the infection. Since SBV has spread to all over the country, efforts have been made to treat and prevent this devastating disease although no effective results have so far been obtained. Several studies have demonstrated that Apis mellifera bee colonies that express an efficient hygienic behavior exhibit a higher resis tance to the brood disease. In this study we demonstrated that the differences of hygienic behavior between A. cerana and A. mellifera. A. cerana more efficiently removed the pin-killed brood than A. mellifera. On the other hand, A. mellifera more efficiently removed SBV-infected larvae and SBV-dead brood than A. cerana. However, it remains unclear whether the advantage of hygienic bee could have efficacy against Sacbrood disease on A. cerana colonies.

Pollinating Characteristics of Apis mellifera and Bombus terrestris on Mango Flower in Vinyl-House Condition

In Gyun Park,Mi Ae Kim,Hyung Joo Yoon,Kyeong Yong Lee,Kyeong San Choi,Yoon Mi Kim 한국응용곤충학회 2013 한국응용곤충학회 학술대회논문집 Vol.2013 No.04

To substitute for bluebottle fly, Chrysomyia megacephala which is being used for pollinator in mango fruit, and improve the pollinating effect of mango fruit which is also being increased as high value added crop recently in Jeju island of korea, 2 kinds of pollinator were used in analyzing and surveying of foraging activities on mango fruit in Seogwipo province. This study was conducted using 3 species of pollinator, Apis mellifera, Bombus terrestris and Chrysomyia megacephala with 3 treatment in vinyl-house condition respectively. Species of mango fruit, Irwin, was used in this experiment. A number of foraging activity of Apis mellifera and Bombus terrestris in hive showed highest 11 AM, and showed normal foraging activity in high temperature condition(28℃). Pollinating ratio of Bombus terrestris was shown 100% and over 95% in case of Apis mellifera. This ratio suggest that the 2 species of insects is effective as pollinator on mango fruit compared with bluebottle fly. Daily pollinating activity of Apis mellifera and Bombus terrestris was shown peak in 11 AM, but showed even activity from 9 AM to 3 PM in case of Chrysomyia megacephala. The pollinating characteristics of 3 species depends on illuminance but temperature, especially in case of Bombus terrestris was more affected by change of illuminance. Visiting time of Bombus terrestris and Apis mellifera on this flower was shown 2.8 and 3.4 seconds respectively. But Chrysomyia megacephala showed longer 10 times with 32.5 seconds than other insects. This results suppose that Chrysomyia megacephala showed as resting behavior for almost time on the flower not foraging activity to pollinate.

Do neonicotinoid insecticides impaired olfactory learning behavior in Apis mellifera?

Imran, Muhammad,Sheikh, Umer Ayyaz Aslam,Nasir, Muhammad,Ghaffar, Muhammad Abdul,Tamkeen, Ansa,Iqbal, Muhammad Aamir Korean Society of Sericultural Science 2019 International Journal of Industrial Entomology Vol.38 No.1

Bee's population is declining and disappearing at alarming rate. There are many factors responsible for declining the population of bees including diseases, natural enemies, environmental conditions and pesticides. Insecticides play its role dramatically for their population decline and neonicotinoid insecticides are critically important due to their wide application for pest control. Keeping in view of above problem, effect of neonicotinoid insecticides on olfactory learning behavior in Apis mellifera was observed using Proboscis Extension Reflex (PER) method. In this method, bees were harnessed in centrifuges tubes and feed on insecticides mixed sugar solution after three hours hunger. Bees were checked by feeding on non-treated sugar solution to observe PER response. Minimum proboscis extension was observed for acetamiprid and imidacloprid with 26% and 20% respectively at their recommend field doses while it was maximum for dinotefuran and thiamethoxam with 73% and 60% respectively. Only 40% bees showed response when exposed at 1/10 concentration of field dose for imidacloprid and the least at 1/100 of field dose. At control (Sugar solution) about 90% bees showed PER response. Among these neonicotinoid insecticides tested, imidacloprid and acetamiprid were the most damaging which impaired the olfactory learning performance in Apis mellifera.

Do neonicotinoid insecticides impaired olfactory learning behavior in Apis mellifera?

Muhammad Imran,Umer Ayyaz Aslam Sheikh,Muhammad Nasir,Muhammad Abdul Ghaffar,Ansa Tamkeen,Muhammad Aamir Iqbal 한국잠사학회 2019 International Journal of Industrial Entomology Vol.38 No.1

Bee’s population is declining and disappearing at alarming rate. There are many factors responsible for declining the population of bees including diseases, natural enemies, environmental conditions and pesticides. Insecticides play its role dramatically for their population decline and neonicotinoid insecticides are critically important due to their wide application for pest control. Keeping in view of above problem, effect of neonicotinoid insecticides on olfactory learning behavior in Apis mellifera was observed using Proboscis Extension Reflex (PER) method. In this method, bees were harnessed in centrifuges tubes and feed on insecticides mixed sugar solution after three hours hunger. Bees were checked by feeding on non-treated sugar solution to observe PER response. Minimum proboscis extension was observed for acetamiprid and imidacloprid with 26% and 20% respectively at their recommend field doses while it was maximum for dinotefuran and thiamethoxam with 73% and 60% respectively. Only 40% bees showed response when exposed at 1/10 concentration of field dose for imidacloprid and the least at 1/100 of field dose. At control (Sugar solution) about 90% bees showed PER response. Among these neonicotinoid insecticides tested, imidacloprid and acetamiprid were the most damaging which impaired the olfactory learning performance in Apis mellifera. Bombyx mori silkworm has many silkworm varieties and the structure and properties of silk can be varied depending on the silkworm variety. Therefore, the choice of proper silkworm variety can be an effective way to control and improve the performance of silk materials in biomedical and cosmetic applications. As a preliminary study, in the present study, thirty nine Bombyx mori silkworm cocoons were prepared and their molecular conformation and crystallinity were examined. Molecular conformation and the crystallinity of silkworm cocoons were different depending on the silkworm variety. Interestingly, the crystallinity index of outside of cocoon was different from that of inside of cocoon and the difference between the crystallinities of outside and inside of cocoon was varied depending on the silkworm variety. In this study, the silkworm variety cocoons were classified into three groups (Group 1, Group 2, and Group 3) based on the difference of crystallinities between outside and inside.

꿀벌 Apis mellifera에서 유래 한 특성화 되지 않은 항균성 펩티드의 동정

박희근(Park Hee Geun),김동원(Dong Won Kim),이만영(Man-Young Lee),최용수(Yong Soo Choi) 한국생명과학회 2020 생명과학회지 Vol.30 No.1

꿀벌(Apis mellifera)에는 많은 항균성 펩티드가 있습니다. 그러나 아직 많은 종류의 펩티드를 기능을 알려지지 않았다. 따라서, 알려지지 않은 기능성 펩티드의 특성화가 필요하다. 그래서 우리는 새로운 항균성 펩티드(AMP)를 분석 하였다. 우리는 Apis mellifera에서 total RNA를 분리하고 Illumina HiSeq 2500 차세대 시퀀싱(NGS) 기술을 사용하여 15,314 개의 펩티드 서열을 생성하여 새로운 AMP를 선발 하였다. AMP로서 기능을 가지는 AMP를 선발 하기 위해 AMP 서열의 특성 과 특징을 분석을 기초로 하여 알려지지 않은 펩티드 및 알려진 44 개의 펩티드가 확인 되었다. 그 중에서도 AMP5라는 특성화 되지 않은 펩티드를 선발 하였다. AMP5는 표피, 지방체, 독낭에서 발현된다. 항균 활성을 분석하기 위해 Gram-negative bacteria Escherichia coli KACC 10005 및 Bacillus thuringiensis KACC 10168에 대한 항균 활성을 합성한 AMP5 처리하여 시험 하였다. AMP5는 Gram-negative bacteria Escherichia coli에 대한 항균 활성을 나타냈다(MIC50 = 22.04±0.66 μM). 일벌에 Escherichia coli을 주사했을 때 AMP5는 체내에서 항균성 펩티드로 발현이 높아졌다. 이러한 결과는 Escherichia coli에 대한 항균 활성을 나타냄을 확인하였다. The European honeybee (Apis mellifera L.) has multiple anti-microbial peptides, but many were unknown and demands for their characterization have increased. This study therefore focused on identifying novel anti-microbial peptides (AMPs) from A. mellifera L. To obtain high-throughput transcriptome data of the honeybee, we implemented next-generation sequencing (NGS), isolating novel AMPs from total RNA, and generated 15,314 peptide sequences, including 44 known, using Illumina HiSeq 2500 technology. The uncharacterized peptides were identified based on specific features of possible AMPs predicted in the sequencing analysis. AMP5, one such uncharacterized peptide, was expressed in the epidermis, body fat, and venom gland of the honeybee. We chemically synthesized this peptide and tested its anti-bacterial activity against Gram-negative Escherichia coli (KACC 10005) and Gram-positive Bacillus thuringiensis (KACC 10168) by anti-microbial assay. AMP5 exhibited anti-bacterial activity against E. coli (MIC50=22.04±0.66 μM) but not against B. thuringiensis. When worker bees were injected with E. coli, AMP5 was up-regulated in the body fat. This study therefore identified AMP5 in adult European honeybees and confirmed its anti-bacterial activity against Gram-negative E. coli.

Effect of royal jelly on longevity and memory-related traits of Apis mellifera workers

Jing-liang Shi,Chun-hua Liao,Zi-long Wang,Xiaobo Wu 한국응용곤충학회 2018 Journal of Asia-Pacific Entomology Vol.21 No.4

Royal jelly (RJ) is a key factor for honey bee caste determination. The queen bee is fed with RJ by worker bees throughout her life, while the worker bees eat bee bread themselves. This study was designed to explore the effect of nutrient-rich RJ on longevity and learning and memory abilities of workers of the western honey bee Apis mellifera. The newly emerged worker bees were randomly divided into three groups and were fed 50% sucrose solution containing 0%, 10%, and 20% RJ. We found that worker bees fed with 10% and 20% RJ showed significantly improved longevity and higher proboscis extension response success rate compared to bees fed with 50% sucrose containing 0% RJ. Additionally, bees fed with 20% RJ showed significantly higher level of expression of memory related genes (GluRA and Nmdar1) compared to the control group. Furthermore, expression of the Nmdar1 gene of worker bees fed with 10% RJ was also significantly higher than in the control group. These results indicate that RJ has potential effects on the longevity and learning and memory abilities of A. mellifera.

Molecular cloning and characterization of a growth-blocking peptide from the honeybee Apis mellifera

Myung Kim Jin,Hui Kim Yun,Ji Park Min,Yeon Kim Bo,Joo Yoon Hyung,Yong Kyeong Lee,Soo Yong Choi,Young Man Lee,Sik Lee Kwang,Rae Jin Byung 한국응용곤충학회 2022 Journal of Asia-Pacific Entomology Vol.25 No.2

Growth-blocking peptides (GBPs) are insect cytokines that have been characterized from several species. How ever, molecular studies on hymenopteran GBPs remain limited. In this study, we cloned four GBP cDNAs of honeybees and bumblebees and compared their sequences with those of other insect GBPs. Sequence analysis revealed that GBP from bees conserves a mature peptide with 21–25 amino acids, including two conserved cysteine residues. During Apis mellifera development, AmGBP was expressed at the larval stage but not at the pupal stage. AmGBP transcript reduction by RNAi affected larval growth and reduced body weight gain. AmGBP RNAi-treated larvae on day 4 were arrested to the larval stage and finally died. Our data suggested that AmGBP plays an essential role in larval growth and acts as a growth factor at the larval stage of A. mellifera.

꿀벌(Apis mellifera)과 stingless bee (Lepidotrigona ventralis flavibasis)의 농약의 급성 독성 비교

정성민,이창열,정철의 한국응용곤충학회 2015 한국응용곤충학회 학술대회논문집 Vol.2015 No.04

꿀벌은 화분매개 산업으로 가장 각광을 받고 있는 곤충 중 하나이다. 현재는 서양뒤열벌, 머리뿔가위벌 등이 화분매개로 많이 이용되며 앞으로 다른 화분매개 곤충을 연구하여 화분매개 산업을 발전 시켜나가 농가와의 시너지효과를 기대할 수 있다. 현대에 들어 화분매개 산업에 문제시 되는 농약의 독성에 대한 피해가 늘어나고 있는데 미래의 화분매개충이 될 확률이 높은 stingless bee와 꿀벌간의 급성 독성에 대한 차이를 알고자 태국 치앙마이 대학교에서 stingless bee를 포획하여 네오니코티노이드계 3종(Thiamethoxam, Imidacloprid, clothianidin과 카바메이트계 1종(Cabaryl)에 대해서 섭식독성 실험을 하였다. 농약별 추천 농도에 맞게 희석 한 후, 10배부터 100,000배 6단계로 나누어 처리 하였다. Stingless bee의 반수치사농도는 Thiamethoxam, Imidacloprid, clothianidin에 대해 각각 049, 5.84, 0.72 ppm이었고, Cabaryl 처리구에서는 처리후 14시간까지 추천농도에서도 20%대의 사망률을 보였다. 반면 A. mellifera의 반수치사농도는 각각 0.22, 1.97, 0.46, 53.6 ppm로 나타났다. A. mellifera 국내 개체군을 대상으로 한 연구에서는 0.19, 6.32, 0.22, 7.84 ppm으로 나타났다. 꿀벌의 경우 개체군간 차이는 매우 적었으며, stingless bee의 경우 독성이 상대적으로 낮게 나타났다.