• Fisheries and Aquatic Sciences
• /
• v.17 no.1
• /
• pp.105-113
• /
• 2014

Glufosinate-ammonium, a component of the herbicide Basta, is one of the most extensively used pesticides worldwide. In this study, we assessed subchronic and chronic toxicities of Basta and its histopathological effects on the marine medaka Oryzias dancena. Marine medaka were exposed to 0, 2, 4, or 8 mg/L of Basta for 28 or 42 days. The lethal concentration ($LC_{50}$) of Basta for 96 h is 8.76 mg/L. Histological changes in the gills and liver were evaluated with histopathological indices, allowing quantification of the damage to fish exposed to Basta. Blood congestion, lamellar fusion, and epithelial lifting were observed in the gills, and hydropic degeneration, fibrosis, lipid degeneration, leukocyte infiltration, and necrosis were found in the liver. These responses could be useful indicators of Basta toxicity in this species.

• Journal of Environmental Impact Assessment
• /
• v.22 no.2
• /
• pp.125-134
• /
• 2013

Plant virus coat (CP) gene-mediated protection is one of the best known approaches to protect against virus resistant transgenic plants. Transgenic N. benthamiana plants containing the CP gene of Zucchini green mottle mosaic virus (ZGMMV) were used for the environmental risk assessment of the living modified (LM) plants with plant virus resistance. The most optimal co-infection method of both ZGMMV and CMV (Cucumber mosaic virus) on Non-LM and CP-expressing LM tobacco plants was established and co-infection of CMV and ZGMMV was confirmed by polymerase chain reaction (PCR). To address the effects of LM tobacco plants on the mutation of the virus, in-vitro transcripts of CP and Replicase (Rep) derived from CMV and/or ZGMMV were inoculated onto Non-LM or LM tobacco plants. Mutation frequency of CP and Rep from CMV and ZGMMV was examined through six serial passages in Non-LM and LM tobacco plants. Little actual frequency of mutation was estimated, probably due to the limited number of transgenic plants tested in this study. However, it does not suggest environmental safety of these CP-mediated LM plants. Further study at a larger scale is needed to evaluate the environmental risk associated with the CP-expressing LM plants.

• Entomological Research
• /
• v.48 no.6
• /
• pp.533-539
• /
• 2018

The RNA interference (RNAi) has been considered as an important genetic tool and applied to develop a new living modified (LM) crop trait which is an improvement of nutrient quality or pest management. The RNAi of DvSnf7 has been used for resistance to LM maize and the Western Corn Rootworm which is a major agricultural pest for the US Corn Belt. Most of the environmental risk assessments (ERA) of double strand RNA (dsRNA) have been performed using in vitro transcript products, and not in vivo expressed product. A large amount of dsRNA was required for the acute toxicity assay of water fleas. Therefore development of massive dsRNA purification techniques is critical. Daphnia, a freshwater microcrustacean, is a model organism for studying cellular and molecular mechanism involved in life history traits and ecotoxicology. In this study, we established the massive dsRNA purification method using Escherichia coli and implemented acute toxicity assays to Daphnia magna. As a result, the present RNase A and DNase I, dsRNA was efficiently purified without any special techniques or equipment. Even though purified dsRNA existed during the acute toxicity test, lethality or abnormal behavior were not observed in D. magna. These results indicated that GFP and DvSnf7 dsRNA were not significantly affected to D. magna due to their lack of sequence matching in its genome. The purification method of dsRNA and the acute toxicity assay of water fleas using purified dsRNA would be suitable for the toxicological studies of LMOs to aquatic non-target organisms.

• Proceedings of the National Institute of Ecology of the Republic of Korea
• /
• v.2 no.2
• /
• pp.129-138
• /
• 2021

Living modified organisms (LMOs) are managed by seven government agencies according to their use in South Korea. The Ministry of Environment is responsible for LMOs used for environmental remediation. This study aimed to develop guidelines for assessing potential risks posed by transgenic plants used for remediation to insect ecosystems by investigating arthropod communities in sunflower fields. A total of 2,350 insects and spiders belonging to 134 species of 10 orders and 71 families were collected from sunflower fields over four growth stages ranging from anthesis to seed maturity. At the R3 phase of flower-bud formation, Chironomidae sp. of a decomposer insect guild presented the highest density, while Apis mellifera of a pollinator guild was the most abundant in the R5.8 phase of flowering. During the R7 seed-filling phase and the R9 phase of seed maturity, herbivorous Pochazia shantungensis predominated. During the R9 phase, richness and diversity indices of arthropod communities were distinctly lower whereas their dominance indices were significantly higher than those at other phases. In addition, the composition of arthropod communities was strongly correlated not only with the sampling date, but also with the sampling method depending on the growth stage of sunflowers. Our results suggest that appropriate sampling timing and methods should be considered in advance and that long-term field trials that cover a variety of environmental conditions should be carried out to evaluate potential risks to insect ecosystems.

• Proceedings of the National Institute of Ecology of the Republic of Korea
• /
• v.2 no.3
• /
• pp.210-218
• /
• 2021

Recently, pest-resistant living modified (LM) crops developed using RNA interference (RNAi) technology have been imported into South Korea. However, the potential adverse effects of unintentionally released RNAi-based LM crops on non-target species have not yet been reported. Coccinella septempunctata, which feeds on aphids, is an important natural enemy insect which can be exposed to the double-stranded RNA (dsRNA) produced by RNAi-based LM plants. To assess the risk of ingestion of Snf7 dsRNA by C. septempunctata, we first identified the species through morphological analysis of collected insects. A method for species identification at the gene level was developed using a specific C. septempunctata 12S rRNA. Furthermore, an experimental model was devised to assess the risk of Snf7 dsRNA ingestion in C. septempunctata. Snf7 dsRNA was mass-purified using an effective dsRNA synthesis method and its presence in C. septempunctata was confirmed after treatment with purified Snf7 dsRNA. Finally, the survival rate, development time, and dry weight of Snf7 dsRNA-treated C. septempunctata were compared with those of GFP and vATPase A dsRNA control treatments, and no risk was found. This study illustrates an effective Snf7 dsRNA synthesis method, as well as a high-concentration domestic insect risk assessment method which uses dsRNA to assess the risk of unintentional released of LM organisms against non-target species.

• Korean Journal of Ichthyology
• /
• v.18 no.2
• /
• pp.65-77
• /
• 2006

Gene transcripts potentially responsive to the heat stress were surveyed by cDNA microarray analysis in mud loach (Misgurnus mizolepis). Transcriptional profiles of hepatic tissue in the fish exposed to either $23^{\circ}C$ or $32^{\circ}C$ for 4 weeks were compared each other by 3 replicated hybridization assays using 1,124 unigene clones selected from mud loach liver expressed sequence tags (ESTs). A total of 93 clones showed the substantially increased mRNA levels (>2-fold) in $32^{\circ}C$-exposed group when compared in $23^{\circ}C$control group. It includes various enzymes and proteins involved in energy pathway, protease/protein metabolisms, immune/antioxidant functions, cytoskeleton/cell structure, transport and/or signal transduction. Maximum level of increase was up to 15-fold relative to $23^{\circ}C$ treatment. Heat exposure also resulted in the significant decrease (less than 50% relative to $23^{\circ}C$-exposed fish) of the transcriptional activities in 85 genes. Besides the above categories, yolk protein (vitellogenin) and ribosomal proteins were notably down regulated in the fish exposed to heat stress. A number of novel gene transcripts were also detected in both up-regulated and down-regulated groups.

• Journal of Aquaculture
• /
• v.19 no.3
• /
• pp.166-172
• /
• 2006

Induced androgenesis, a form of artificial parthenogenesis is an important tool for the generation and use of genetically isogenic or clonal fish strain. An optimized protocol for the genetic inactivation of mud loach (Misgurnus mizolepis) oocytes (i.e. production of androgenetic haploid) was developed using UV-irradiation. Various dose levels of UV significantly affected the fertilizing capacity of the eggs, hatchability of embryos and incidence of haploidy. Based on the extensive examinations of treatment conditions on embryo viability and haploid incidence, the optimum dose level of UV irradiation was turned out to be $10,800ergs/mm^2$ with 56.9% of hatching success and 94.6% of haploidy. The overall yield of putative androgen under optimized treatment condition was more than 50% out of total eggs inseminated. The success of androgenetic reproduction of haploid genome was verified by flow cytometry and PCR amplification of transgene that is exclusive to either one of parental sexes. However, a small portion $(8\sim11%)$ of presumed androgenetic haploid larvae was proven to contain residual DNA fragment(s) from maternal parent.

• Journal of Plant Biotechnology
• /
• v.44 no.1
• /
• pp.97-106
• /
• 2017

Living modified crops are genetically modified living organisms and are widely used in biotechnical research and desired goods. As the reliance on LM products, concerns about safety of LMOs have been continuously increased in South Korea. We established the detection methods for unintentional released LMOs in environmental conditions. To detect six LM event genes of 1 canola, 1 maize and 4 soybeans, PCR conditions were based upon consideration of the Joint Research Centre information. Genomic DNAs were isolated from LM samples and PCR analysis were performed using each event-specific primer pair. Event-specific genes of all events were efficiently recognized by our methods. To investigate the insertion site of LM genes in each genome, we verified PCR product sequence by DNA sequencing. These results suggest that the LM event-specific gene amplification can be efficiently developed. In addition, our detection method is fit for monitoring and post-management of LM crops in the environment.

• Journal of Plant Biotechnology
• /
• v.42 no.3
• /
• pp.196-203
• /
• 2015

AbstractLiving modified organisms (LMO) are one of the most widespread products of modern biotechnology after DNA discovery. Due to the decline of grain self-sufficiency rate and the increase of reliance on LMO imports in Korea, a series of concerns with regard to safety of living modified(LM) crops has been raised. The aim of this study is to establish the detection methods for unintentional release or growing of LMO plants in environmental conditions. To detect LM crop events, general concepts of specific primer design and PCR conditions were provided by the Joint Research Centre (JRC). The certified reference materials of seven LM events (4 soybean, 2 cotton and 1 corn) were obtained from the Institute for Reference Materials and Measurements (IRMM) and the American Oil Chemists' Society (AOCS). Genomic DNA from seven LM events were purified and PCR amplifications were carried out by using individual event-specific primer sets. LM-specific PCR products of all seven events were efficiently amplified by our methods. The results indicate that the established detection method for LMOs is suitable as a scientific tool to monitor whether the crops found in natural environments are LMOs.

• Korean Journal of Environmental Biology
• /
• v.39 no.4
• /
• pp.415-422
• /
• 2021

Cotton is an important fiber crop, and its seeds are used as feed for dairy cattle. Crop biotechnology has been used to improve agronomic traits and quality in the agricultural industry. The frequent unintentional release of LM cotton into the environment in South Korea is attributed to the increased application of living modified (LM) cotton in food, feed, and processing industries. To identify and monitor the LM cotton, a method for detecting the approved LM cotton in South Korea is required. In this study, we developed a method for the simultaneous detection of four LM cotton varieties, MON757, MON88702, COT67B, and GHB811. The genetic information of each LM event was obtained from the European Commission-Joint Research Centre and Animal and Plant Quarantine Agency. We designed event-specific primers to develop a multiplex PCR method for LM cotton and confirmed the specific amplification. Using specificity assay, random reference material(RM) mixture analysis and limit of detection(LOD), we verified the accuracy and specificity of the multiplex PCR method. Our results demonstrate that the method enabled the detection of each event and validation of the specificity using other LM RMs. The efficiency of multiplex PCR was further verified using a random RM mixture. Based on the LOD, the method identified 25 ng of template DNA in a single reaction. In summary, we developed a multiplex PCR method for simultaneous detection of four LM cotton varieties, for possible application in LM volunteer analysis.