• Korean Journal of Microbiology
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• v.49 no.3
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• pp.270-274
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• 2013

The bumblebee, Bombus terrestris, has played an important role as one of the alternative pollinators since the outbreak of honeybee collapse disorder. Recently, pathogens and parasites such as viruses, bacteria and mites, which affect the life span and fecundity of their host, have been discovered in B. terristris. In order to detect the microsporidian pathogen, Nosema spp. in the field populations of B. terristris, we collected adults and isolated their genomic DNA for diagnostic PCR. The PCR primers specific for Nosema spp. were newly designed and applied to gene amplification for cloning. Only small subunit ribosomal RNA (SSU rRNA) gene of N. ceranae was successfully amplified among examined genes and sequenced, which indicates that N. ceranae mainly infects the examined field population of B. terristris. To detect of SSU rRNA gene, two regions of SSU rRNA gene were selected by primary PCR analysis and further analyzed in quantitative real-time PCR (qRT-PCR). The qRT-PCR analysis demonstrated that SSU rRNA of N. ceranae was detected at concentration as low as $0.85ng/{\mu}l$ genomic DNA. This result suggests that the detection via qRT-PCR can be applied for the rapid and sensitive diagnosis of N. ceranae infection in the field population as well as risk assessment of B. terristris.

• Journal of Apiculture
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• v.32 no.3
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• pp.147-154
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• 2017

Honeybees (Apis mellifera) are common pollinators and important insects studied in agriculture, ecology and basic research. Recently, RDA (Rural Development Administration) and YIRI (Yecheon-gun Industrial Insect Research Institute) have been breeding a triple crossbred honey bee named Jangwon, which have the ability to produce superior quality honey. In this study, we identified a single nucleotide polymorphism (SNP) marker in the genome of Jangwon honeybee, particularly, in the paternal line (D line). Initially, we performed Sequence-Based Genotyping (SBG) using the Illumina Hiseq 2500 in 5 honeybee inbred lines; A, C, D, E, and F; and obtained 1,029 SNPs. Seventeen SNPs for each inbred line were generated and selected after further filtering of the SNP dataset. The 17 SNP markers validated by performing TaqMan probe-based real-time PCR and genotyping analysis was conducted. Genotyping analysis of the 5 honeybee inbred lines and one hybrid line, $D{\times}F$, revealed that one set of SNP marker, AmD9, precisely discriminated the inbred line D from the others. Our results suggest that the identified SNP marker, AmD9, is successful in distinguishing the inbred honeybee lines D, and can be directly used for genotyping and breeding applications.

• Korean Journal of Environmental Agriculture
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• v.36 no.4
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• pp.241-248
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• 2017

BACKGROUND: RNA interference (RNAi) eliminates or decreases gene expression by disrupting the target mRNA or by interfering with translation. Recently, RNAi technique was applied to generate new crop traits which provide protection against pests. To establish the environmental risk assessment protocol of RNAi LMO in lab scale, we developed dsRNA expression system using E. coli and tested acute oral toxicity assay to honey. METHOD AND RESULTS: The dsRNA expression vector, L4440, was chosen and cloned 240 bp of Snf7 and GFP gene fragment. To develop the maximum dsRNA induction condition in E. coli, we tested induction time, temperature and IPTG concentration in media. To estimate the risk assessment of dsRNA to honey bee, it has been selected and cultured with dsRNA supplement for 48 hours according to OECD guideline. As a result, the optimum condition of dsRNA induction was $37^{\circ}C$, 4 hours and 0.4 mM IPTG concentration and the difference between Snf7 and GFP dsRNA molecules from E. coli was not significant in survival and behavior to honey bee. Furthermore, blast search results indicated that effective match of predicted dsRNA fragments were not existed in honey bee genome. CONCLUSION: In this study, we developed and tested the acute oral toxicity of dsRNA using E. coli expression system to honey bee.

• Korean Journal of Environment and Ecology
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• v.33 no.3
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• pp.303-320
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• 2019

Plants interact with various biotic and abiotic environmental factors. It requires much information to understand the traits of a plant species. A shortage of information would restrict the assessment, especially in the evaluation of what kind of factors influence a plant species to face extinction. Polygonatum stenophyllum Maxim. is one of the northern plants of which Korea is the southern distribution edge. The Korean Ministry of Environment had designated it to be the endangered species until December 2015. Although it is comparatively widespread, and a large population has recently been reported, it is assessed to be vulnerable due to the low population genetic diversity. This study evaluated the current distribution of Polygonatum stenophyllum Maxim. We investigated the vegetational environment, population structures, phenology, soil environment, and self-incompatibility based on the results. Lastly, we evaluated the current threats observed in the habitats. The habitats tended to be located in the areas where the masses at the edge of the stream accumulated except for those that were located on slopes of some mountainous areas. Most of them showed a stable population structure and had re-established or recruited seedlings. Polygonatum stenophyllum Maxim. had the difference in time when the shoots appeared above the ground depending on the depth of the rhizome located in the underground. In particular, the seedlings and juveniles had their rhizome located shallow in the soil. Visits by pollinator insects and success in pollination were crucial factors for bearing of fruits by Polygonatum stenophyllum Maxim. The threats observed in the habitat of Polygonatum stenophyllum Maxim. included the expansion of cultivated land, construction of new buildings, and construction of river banks and roads. Despite such observed risk factors, it is not likely that there would be rapid population reduction or extinction because of its widespread distribution with the total population of more than 2.7 million individuals and the new populations established by the re-colonization.

• Korean Journal of Environment and Ecology
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• v.33 no.4
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• pp.422-439
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• 2019

The rear edge population is considered to have low genetic diversity and high risk of extinction according to a highly isolated distribution. However, the rear edge population is observed to have persisted for an extended period despite the low genetic diversity. As such, it is necessary to understand the ecological process involved in the persistence of the population. Viola mirabilis L. in Korea is considered the rear edge population from the perspective of the worldwide distribution. We surveyed the distribution range of V. mirabilis, which shows the isolated distribution in the central area of Korea, to find out the factors of its persistence. Next, we investigated and accessed the vegetational pattern of habitats, soil environment, phenology, self-compatibility, population structure, and extinction risk factors observed in the distribution area. V. mirabilis was distributed in the understory of the deciduous forest, planted forest of the deciduous conifer and deciduous broad-leaved trees, shrubland, and grassland in the limestone area. We also observed the re-establishment of seedlings in the population, and most of them showed a stable population structure. For chasmogamous flowers, the visit by pollinators has a significantly positive relationship with the production of fruits. However, we found that the production of the cleistogamous flowers was more numerous in all studied populations and that only the cleistogamous flowers were produced despite a more substantial plant size in some populations. The plant size was more related to the production of the cleistogamous flowers than that of the chasmogamous flowers. Accordingly, the cleistogamous flowers significantly contributed to seedling recruitment in the population. We found that the production of the chasmogamous flowers and the cleistogamous flowers did not have a correlation with the factors of the soil analysis except for phosphoric acid. V. mirabilis showed the self-incompatibility characteristics most likely due to the production capability of the cleistogamous flowers. Potential extinction risk factors observed in the distribution area was included the development of limestone mine, the expansion of agricultural fields, and the construction of houses. Although V. mirabilis showed an isolated distribution in the limestone area in the Korean peninsula, it showed a diverse distribution in a wide habitat environment ranging from the grassland to the understory of the trees with relatively low canopy closure rate. Moreover, we concluded that the persistence of the population was possible if we can maintain the current state of multiple populations and stable population structure.

• Korean Journal of Environment and Ecology
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• v.36 no.3
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• pp.303-317
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• 2022

Climate change caused by increased greenhouse gas emissions can alter the natural ecosystem, including the pollination ecosystem and agricultural ecology, which are ecological interactions between potted insects and plants. Many studies have reported that populations of wild bees, including bees and wasps (BW), which are the key pollinators, have gradually declined due to climate change, leading to adverse impacts on overall biodiversity, ultimately with agribusinesses and the life cycle of flowering plants. Therefore, we could infer that the rising temperature in Korean Peninsula (South Korea) due to global warming has led to climate change and influenced the wild bee's ecosystem. In this study, we surveyed the distributional pattern of BW (Superfamily: Apoidea, Vespoidea, and Chrysidoidea) at 51 sites from 2017 (37 sites) to 2018 (14 sites) to examine the effects of climatic factors on the nationwide distribution of BW in South Korea. Previous literature has confirmed that their distribution according to forest climate zones is significantly correlated with mean and accumulative temperatures. Based on the result, we predicted the effects of future climate changes on the BW distribution that appeared throughout South Korea and the species that appeared in specific climate zones using Shared Socioeconomic Pathways (SSPs). The distributions of wild BW predicted by the SSP scenarios 2-4.5 and 5-8.5 according to the BIOMOD species distribution model revealed that common and endemic species will shift northward from the current habitat distribution by 2050 and 2100, respectively. Our study implies that climate change and its detrimental effect on the ecosystem is ongoing as the BW distribution in South Korea can change, causing the change in the ecosystem in the Korean Peninsula. Therefore, immediate efforts to mitigate greenhouse gas emissions are warranted. We hope the findings of this study can inspire further research on the effects of climate change on pollination services and serve as the reference for making agricultural policy and BW conservation strategy

• The Korean Journal of Pesticide Science
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• v.13 no.1
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• pp.39-44
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• 2009

This study was conducted to evaluate the actual risk of fipronil on worker honey bees (Apis mellifera L.) through acute contact toxicity test, acute oral toxicity test, toxicity of residues on foliage test, and small scale field test. The $48h-LD_{50s}$ of fipronil SC on honeybee were $0.005{\mu}g$ a.i./bee in acute contact toxicity test and $0.004{\mu}g$ a.i./bee in acute oral toxicity test, respectively. In toxicity of residues on foliage test, fipronil showed over 90% of mortality during 28days after treatment at recommended application rate. The $DT_{50}$ of dislodgeable foliar residue was 9 days. Finally, In small scale field test, fipronil showed similar toxicity in the residues on foliage test. It was concluded that fipronil has very high acute toxicity and long residual toxicity to honeybee. Therefore, fipronil is highly toxic to bees exposed to direct treatment or residues on blooming crops or weeds. Do not apply this product or allow it to drift to blooming crops or weeds if bees are visiting the treatment area. To protect honeybee and wild pollinators from outdoor use of fipronil, ultimately it should need to limit for only indoor use to prevent pollinators from unintentionally exposure of fipronil.