• Journal of Plant Biotechnology
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• 제47권4호
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• pp.309-315
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• 2020

Advances in biotechnology have led to progress in crop genetic engineering to improve agricultural productivity. The use of genetically modified (GM) crops has increased, as have consumers' and regulators' concerns about the safety of GM crops to human health, and ecological biodiversity. As such, the identification of GM crops is a critical issue for developers and distributors, and their labeling is mandatory. Multiplex polymerase chain reaction (PCR) has been developed and its use validated for the detection and identification of GM crops in quarantine. Herein, we established a simultaneous detection method to identify four GM maize events. Event-specific primers were designed between the junction region of transgene and genome of four GM maize lines, namely 5307, DAS-40278-9, MON87460, and MON87427. To verify the efficiency and accuracy of the multiplex PCR we used specificity analysis, limit of detection evaluation, and mixed certified reference materials identification. The multiplex PCR method was applied to analyze 29 living, modified maize volunteers collected in South Korea in 2018 and 2019. We performed multiplex PCR analysis to identify events and confirmed the result by simplex PCR using each event-specific primer. As a result, rather than detecting each event individually, the simultaneous detection PCR method enabled the rapid analysis of 29 GM maize volunteers. Thus, the novel multiplex PCR method is applicable for living modified organism volunteer identification.

• 농업과학연구
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• 제48권4호
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• pp.1051-1065
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• 2021

Recent times have seen sustained increases in genetically modified (GM) crops not only for cultivation but also for the utility of food and feed worldwide. Domestically, commercial planting and the accidental or unintentional release of living modified (LM) crops into the environment are not approved. Many detection methods had been devised in an effort to realize effective management of the safety of agricultural genetic resources. In order to develop event-specific polymerase chain reaction (PCR) markers for LM crops, we analyzed the genetic information of LM crops. Genetic components introduced into crops are of key importance to provide a basis for the development of detection methods for LM crops. To this end, a total of 18 varieties from four major LM crop species (maize, canola, cotton, and soybeans) were subjected to an analysis. The genetic components included introduced genes, promoters, terminators and selection markers. Thus, if proper monitoring techniques and single or multiplex PCR strategies that rely on selection markers can be established, such an accomplishment can be regarded as a feasible solution for the safe management of staple crop resources.

• Genomics & Informatics
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• 제13권3호
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• pp.81-85
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• 2015

Molecular characterization technology in genetically modified organisms, in addition to how transgenic biotechnologies are developed now require full transparency to assess the risk to living modified and non-modified organisms. Next generation sequencing (NGS) methodology is suggested as an effective means in genome characterization and detection of transgenic insertion locations. In the present study, we applied NGS to insert transgenic loci, specifically the epidermal growth factor (EGF) in genetically modified rice cells. A total of 29.3 Gb (${\sim}72{\times}coverage$) was sequenced with a $2{\times}150bp$ paired end method by Illumina HiSeq2500, which was consecutively mapped to the rice genome and T-vector sequence. The compatible pairs of reads were successfully mapped to 10 loci on the rice chromosome and vector sequences were validated to the insertion location by polymerase chain reaction (PCR) amplification. The EGF transgenic site was confirmed only on chromosome 4 by PCR. Results of this study demonstrated the success of NGS data to characterize the rice genome. Bioinformatics analyses must be developed in association with NGS data to identify highly accurate transgenic sites.

• Journal of Ecology and Environment
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• 제46권3호
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• pp.190-201
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• 2022

Background: The introduction of new living modified (LM) crops may pose a latent threat to the biodiversity of each country. Here, we used sunflowers (Helianthus annuus L.) as a study system to investigate the potential for invasiveness of LM crops under different environmental conditions when released into a natural ecosystem in South Korea. We examined the seed germination, survival, and flowering of sunflowers under competition with wild plants at different sowing dates (March-December) and plot sizes (1 m × 1 m and 2 m × 2 m). Results: The germination rate showed a significant difference according to the sowing date. In addition, several sunflowers survived in plots with a high germination rate, which also led to a higher flowering rate. We found that the smaller the plot, the smaller the area available for inter-species competition, and the higher the number of surviving sunflower plants. The relative dominance and importance value of the species varied significantly between the sowing dates; in particular, sunflowers sown in March could compete with wild plants for longer than those sown on other sowing dates. Conclusions: These observations indicate that the potential for invasiveness of sunflowers differs depending on the environmental conditions and seed density at the time of release.

• 한국작물학회:학술대회논문집
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• 한국작물학회 2001년도 추계 학술대회지
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• pp.144-145
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• 2001

• 환경생물
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• 제39권4호
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• pp.415-422
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• 2021

면화는 중요한 섬유 작물로 종자는 가축의 사료로 사용된다. 작물 생명공학은 농업 분야에서 농업적 형질과 질을 향상시키기 위해 활용되어져 왔다. 국내 식품, 사료, 가공 제품에 유전자변형(LM) 면화의 사용이 증가함에 따라 환경으로의 LM 면화의 비의도적 유출 또한 증가하고 있다. LMO 모니터링 사업에서 수집된 LM 면화를 검정하기 위하여 국내 수입 승인된 LM 면화의 검출법 개발이 필요하다. 본 연구에서는 LM 면화 MON757, MON88792, COT67B, GHB811 4종을 대상으로 동시검출법을 개발하였다. 이벤트에 대한 유전 정보는 유럽 JRC와 농림축산검역본부에서 확보하였다. LM 면화의 동시검출법 개발을 위해 이벤트 특이적인 프라이머를 설계하였으며 특이적인 증폭을 확인하였다. 특이도 검정, 무작위 표준물질 혼합물 분석, 검출한계 분석을 통하여 동시검출법의 정확도와 특이도를 검증하였다. 그 결과 본 동시검출법은 각각의 이벤트를 검출할 수 있으며 LM 표준물질을 활용하여 특이도를 검정하였다. 또한 무작위 표준물질 조합도 정확하게 검출할 수 있다. 검출한계 분석에서는 25 ng의 미량의 주형 DNA로 단회 분석으로 검출이 가능하다. 결론적으로 4종의 LM 면화 동시검출법을 개발하였으며 LM 면화 자생체 분석에 활용될 것으로 사료된다.

• 한국작물학회지
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• 제45권1호
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• pp.59-70
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• 2000

In nature, plant diseases, insects and parasites (hereafter called as "pest") must be co-survived. The most common expression of co-survival of a host crop to the pest can be tolerance. With tolerance, chemical uses can be minimized and it protects environment and sustains host productivity and the minimum pest survival. Tolerance can be applicable in all living organisms including crop plants, lifestocks and even human beings. Tolerant system controls pest about 90 to 95% (this pest control system often be called as horizontal or partial resistance), while the use of chemicals or selection of high resistance controls pest 100% (the most expression of this control system is vertical resistance or true resistance). Controlling or eliminating the pests by either chemicals or vertical resistance create new problems in nature and destroy the co-survial balance of pest and host. Controlling pests through tolerance can only permit co-survive of pests and hosts. Tolerance is durable and environmentally-friend. Crop cultivars based on tolerance system are different from those developed by genetically modified organism (GMO) system. The former stabilizes genetic balance of a pest and a host crop in nature while the latter destabilizes the genetic balance due to 100% control. For three decades, the author has implemented the tolerance system in breeding maize cultivars against various pests in both tropical and temperate environments. Parasitic weed Striga species known as the greatest biological problem in agriculture has even been controlled through this system. The final effect of the tolerance can be an integrated genetic pest management (IGPM) without any chemical uses and it makes co-survival of pests in nature.in nature.

• Journal of Plant Biotechnology
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• 제35권1호
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• pp.31-39
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• 2008

We analyzed the current research trends of living modified organisms (LMO) by questionnaires in the interest of making biosaftey laws and policies in Korea. We executed a pre-survey at the Crop Functional Genomics 2004 conference and obtained LMO research information from 423 LMO research organizations, including 32 national research institutes, 314 universities, and 77 industries. We found that the total 59 kinds of hosts including 26 kinds of plants, 15 kinds of animals, and 18 kinds of microbes were used for LMO research and E. coli was the most common host. The risk of the most experimental hosts was below a biosafety level of 1 (73.8%) and 2 (25.9%). LMO development use purpose was implemented in various developmental uses: 51.3% in test and research use, 19% in health and medical use, and 12.9% in agriculture use. The experiment product, waste product, and products of host for LMO development were 327.2, 223.6, and 13.5 in number of plants; 280.6, 52.4, and 8.7 in number of animals; and $8.3\;{\times}\;10^{11}CFU$, $7.7\;{\times}\;10^{11}CFU$, and $6.5\;{\times}\;10^{11}CFU$ in microbes in 2004. The survey results about how to possess the LMO were very unreliable, because only 10.6% of the researchers returned the questionnaires. Consequently, we strongly suggest the scientific organizations as well as scientists should have more interests in biosafety of LMO research and an LMO biosafety management system should be developed for Korea's future biotechnology.

• Entomological Research
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• 제48권6호
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• pp.533-539
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• 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.

• Journal of Plant Biotechnology
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• 제42권3호
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• pp.196-203
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• 2015

2008년 이후 꾸준히 유전자변형생물체의 수입 승인이 증가하여 2015년 2월 기준 국내 131개의 이벤트가 식용, 사료용, 가공용으로 수입 및 유통 중이다. LMO의 비의도적 자연 생태계 유출을 파악하기 위해 선행되어야 하는 것은 명확한 LMO 도입유전자 검출 기법의 개발이며, 본 연구를 통해 2014년 7개 이벤트에 대한 도입유전자 검출 기법을 확립하였다. 현재까지 확보된 유전자 분석기법은 40개이며 이는 국내도입 LMO의 80%를 검출할 수 있는 과학적 근거를 제공할 것이다(Lee et al. 2015). LMO 자연환경 모니터링 및 사후관리 연구를 위해 단일이벤트를 검출할 수 있는 기법 연구는 앞으로 계속 수행할 계획이다.