• Title/Summary/Keyword: biosafety assessment

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Flanking Sequence and Copy-Number Analysis of Transformation Events by Integrating Next-Generation Sequencing Technology with Southern Blot Hybridization

  • Qin, Yang;Woo, Hee-Jong;Shin, Kong-Sik;Lim, Myung-Ho;Cho, Hyun-Suk;Lee, Seong-Kon
    • Plant Breeding and Biotechnology
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    • v.5 no.4
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    • pp.269-281
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    • 2017
  • With the continual development of genetically modified (GM) crops, it has become necessary to develop detailed and effective molecular characterization methods to select candidate events from a large pool of transformation events. Relative to traditional molecular analysis methods such as the polymerase chain reaction (PCR) and Southern blot hybridization, next generation sequencing (NGS) technology for whole-genome sequencing of complex crop genomes had proven comparatively useful for in-depth molecular characterization. In this study, four transformation events, including one in Bacillus thuringiensis (Bt)-resistant rice, one in resveratrol-producing rice, and two in beta-carotene-enhanced soybeans, were selected for molecular characterization. To merge NGS analysis and Southern blot-hybridization results, we confirmed the transgene insertion sites, insertion construction, and insertion numbers of these four transformation events. In addition, the read-coverage depth assessed by NGS analysis for inserted genes might provide consistent results in terms of inserted T-DNA numbers in case of complex insertion structures and highly duplicated donor genomes; however, PCR-based methods can produce incorrect conclusions. Our combined method provides an effective and complete analytical approach for whole-genome visual inspection of transformation events that require biosafety assessment.

Molecular biological characteristics and analysis using the specific markers of leaf folder-resistant GM rice (혹명나방저항성 GM 벼의 분자생물학적 특성 및 특이 마커를 이용한 검정)

  • Shin, Kong-Sik;Lee, Si-Myoung;Lim, Sun-Hyung;Woo, Hee-Jong;Cho, Hyun-Suk;Lee, Kyeong-Ryeol;Lee, Myung-Chul;Kweon, Soon-Jong;Suh, Seok-Cheol
    • Journal of Plant Biotechnology
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    • v.36 no.2
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    • pp.115-123
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    • 2009
  • In recent years, several genetically modified (GM) crops have been developed worldwide through the recombinant DNA technology and commercialized by various agricultural biotechnological companies. Commercialization of GM crops will be required the assesment of risks associated with the release of GM crops. In advance of the commercial release of GM crops, developer should submit the several information on GM crops for approval. In this study, we carried out to provide the molecular data for the risk assessment of GM rice containing insect-resistant gene, modified Cry1Ac (CryIAc1). Through the molecular analysis with CryIAc1 induced GM rice, we confirmed the steady integration and expression of transgene, the transgene copy number, the adjacent region sequences of inserted gene into rice genome, and the transgene stability in progenies. For the qualitative PCR detection methods, specific primer pairs were designed on the basis of integration sequences, and construct- and event-specific detection markers were developed for leaf folder-resistant rice, Cr7-1 line. From these results, we demonstrated that the molecular data and the PCR detection methods of leaf folderresistant GM rice could be acceptable to conduct the biosafety and environment risk assessment.

Prevalence and Quantification of Vibrio parahaemolyticus in Raw Salad Vegetables at Retail Level

  • Tunung, R.;Margaret, S.P.;Jeyaletchumi, P.;Chai, L.C.;Zainazor, T.C. Tuan;Ghazali, F.M.;Nakaguchi, Y.;Nishibuchi, M.;Son, R.
    • Journal of Microbiology and Biotechnology
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    • v.20 no.2
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    • pp.391-396
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    • 2010
  • The purpose of this study was to investigate the biosafety of Vibrio parahaemolyticus in raw salad vegetables at wet markets and supermarkets in Malaysia. A combination of the most probable number-polymerase chain reaction (MPN-PCR) method was applied to detect the presence of V. parahaemolyticus and to enumerate their density in the food samples. The study analyzed 276 samples of common vegetables eaten raw in Malaysia (Wild cosmos=8; Japanese parsley=21; Cabbage=30; Lettuce=16; Indian pennywort=17; Carrot=31; Sweet potato=29; Tomato=38; Cucumber=28; Four-winged bean=26; Long bean=32). The samples were purchased from two supermarkets (A and B) and two wet markets (C and D). The occurrence of V. parahaemolyticus detected was 20.65%, with a higher frequency of V. parahaemolyticus in vegetables obtained from wet markets (Wet market C=27.27%; Wet Market D=32.05%) compared with supermarkets (Supermarket A=1.64%; Supermarket B=16.67%). V. parahaemolyticus was most prevalent in Indian pennywort (41.18%). The density of V. parahaemolyticus in all the samples ranged from <3 up to >2,400 MPN/g, mostly <3 MPN/g concentration. Raw vegetables from wet markets contained higher levels of V. parahaemolyticus compared with supermarkets. Although V. parahaemolyticus was present in raw vegetables, its numbers were low. The results suggest that raw vegetables act as a transmission route for V. parahaemolyticus. This study will be the first biosafety assessment of V. parahaemolyticus in raw vegetables in Malaysia.

Analysis of risk management system of GM crops in China for the development of global GM crops (글로벌 GM 작물 실용화를 위한 중국의 GM 작물 안전관리제도 분석)

  • Lee, Shin-Woo;Cho, Kwang-Soo;Wang, Zhi;Kwak, Sang-Soo
    • Journal of Plant Biotechnology
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    • v.39 no.3
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    • pp.127-132
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    • 2012
  • We analysed the current status of development of GM crops and national biosafety framework including legislation-related agricultural GMO in China to provide the policy for the development of global GM crops in Korea. In China, several GM crops including cotton, petunia, tomato, sweet pepper, poplar, and papaya have been approved for commercialization and they have been cultivated at more than 4 million ha. In addition, GM rice and GM maize have also obtained approval for productive testing in 2009. China will be the first country to approve GM rice for commercialization. Prior to commercialization in China, all GM crops must be approved by government authority for biosafety assessment specified by national legislation including restricted field testing, enlarged field testing, productive testing and safety certificate. According to China's legislation, agricultural GMOs have been classified by research and testing, production and processing. All GMOs must go through 3 steps of field testing (restricted, enlarged and productive). Prior to conducting each field testing, it has to be approved by government authority. It is assumed that at least one to two years will be taken for each step of field testing (total 4 to 8 years to obtain the final safety certificate) along with a large amount of budget.

Studies on Insect Diversity Related to Genetically Engineered Vitamin A Rice under Large Scale Production (비타민 A 강화 벼의 대규모 GMO 포장에서 곤충다양성 분석)

  • Kim, Kyung-Min;Ryu, Tae-Hun;Suh, Sang Jae
    • Korean Journal of Breeding Science
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    • v.42 no.2
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    • pp.157-162
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    • 2010
  • This study was carried out to develop of macro-protocol and the biosafety guide for 'Nakdong', and 'Golden Rice' (genetically modified vitamin A rice) by large scale field trial of GM crops. Typically, when a new GM crop is created, breeders should conduct field test to make sure the GM crop is safe, and provide some information on GM crops for approval. A total of isolated 4,700 $m^2$ field for trial of GM crops were prepared, and 'Nakdong' and 'Golden rice' were cultivated by standard method of RDA (Rural Development Administration, Korea). Field studies indicated that the population densities of insect pests and natural enemies have no difference between two varieties. While insect pest density on 'Nakdong' was slightly higher than on Golden Rice, but natural enemy density on Golden Rice was a little higher. These results provided the insect diversity for risk assessment analysis of Golden Rice and suggested that the macro-protocol could be useful to detect GM plants.

A Comparative Study of Predicted Environmental Concentrations from ECETOC TRA Based on Environmental Release Categories/Specific Environmental Release Categories and K-CHESAR Using Main/Industrial/Use Categories (환경배출범주/특수환경배출범주 기반 ECETOC TRA 및 주요/산업/용도 분류체계 이용의 K-CHESAR에 의한 환경예측농도 비교 연구)

  • Hyun Pyo Jeon;Jisu Yang;Hana Jo;Eun Kyung Choe;Sanghun Kim
    • Journal of Environmental Health Sciences
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    • v.49 no.6
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    • pp.312-323
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    • 2023
  • Background: Environmental concentrations of substances can be estimated by K-CHESAR based on main, industrial, and use categories (MC/IC/UC) and ECETOC TRA based on environmental or specific environmental categories (ERC or spERC). Objectives: Three different systems for estimating environmental concentrations were compared to figure out their order with possible reasons along with relationship of regional predicted environmental concentrations (PECregional) and final PEClocal for various uses of a substance. Methods: Typical uses of the case substance and their corresponding ERCs were selected from the webpage of the European Chemical Agency. Proper MC/IC/UC and spERC were assigned to each ERC. Emission fractions were compared for each assessment code from the available database. PECs were calculated by three estimating systems: K-CHESAR using MC/IC/UC, ECETOC TRA using ERC, and ECETOC TRA using spERC with their default values for input parameters. Percentage of PECregional to PEClocal were manually calculated for each use. Results: Emission factors decreased in the order of ERC > MC/IC/UC > spERC. Values of the final PEClocal derived as sum of PECregional and Clocal decreased in the order of calculations using ECETOC TRA-ERC>KCHESAR with MC/IC/UC>ECETOC TRA-spERC for all environmental media. Percentages of PECregional,water to PEClocal,water ranged from 0 to 10.3% in industrial uses calculated with MC/IC/UC and ERC but 96.3 to 100% in wide dispersive uses of ERC and spERC where values of Clocal,water are estimated to be very low. Conclusions: ECETOC TRA generated the most refined PNEC values with spERC and the least with ERC, while K-CHESAR with MC/IC/UC generated values between the two results. The ratio of PECregional to PEClocal can be a good measure for performing suitable estimation of PNECs according to use.

Gene flow from herbicide resistant genetically modified rice to conventional rice (Oryza sativa L.) cultivars

  • Han, Sung Min;Lee, Bumkyu;Won, Ok Jae;Hwang, Ki Seon;Suh, Su Jeoung;Kim, Chang-Gi;Park, Kee Woong
    • Journal of Ecology and Environment
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    • v.38 no.4
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    • pp.397-403
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    • 2015
  • Rice (Oryza sativa L.) is an important feeding crop in Asia, and utilization of genetically modified (GM) rice is highly demanding. For co-existence of GM rice and non-GM rice, the proper confinement measures should be provided. Thus, we surveyed gene flow from herbicide resistant GM rice to the conventional rice cultivars in the field tests. Gene flow frequency decreased with increasing distance between the pollen donor and recipients and did not exceed more than 1% even at the nearest distance. In single recipient model plot, a maximum gene flow frequency was observed at the shortest distance and hybrid was detected up to 12 m from the pollen donor. The direction of gene was coincided with the dominant wind direction. Gene flow assessment to multiple recipient plots was conducted under the high raining season by chance, and abrupt decline of gene flow frequency and maximum distance were resulted. According to the survey results, current regulation for isolation distance is reasonable for environmental safety or for general crop production. However, we suggest an alternative measure for GM rice cultivation that should be supplemented to overcome the out of estimation and in the environment asking higher security levels.

Risk assessment of genetically engineered rice Bt-9 resistant to Cnaphalocrocis medinalis: influence on above-ground arthropods in Korea

  • Oh, Sung-Dug;Bae, Eun Ji;Park, Soo-Yun;Lee, Bumkyu;Yun, Do Won;Suh, Sang Jae
    • Korean Journal of Agricultural Science
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    • v.46 no.4
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    • pp.827-841
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    • 2019
  • The effect of genetically engineered rice Bt-9 on the diversity and abundance of plant-dwelling insects and spiders was tested under field conditions. Genetically engineered rice Bt-9, expressing mCry1Ac1 from Bacillus thuringiensis, confers resistance to rice leaf roller (Cnaphalocrocis medinalis) and provides tolerance to the herbicide glufosinate (PPT). The study compared Bt-9 and two non-GM reference varieties, Ilmi-byeo and Dongjin-byeo, at LMO isolated fields in Gunwi (Kyungpook National University) and Jeonju (National Institute Agricultural Sciences) in Southern Korea in 2016 - 2017. A total of 40,817 individuals from 62 families and 11 orders were collected from the two living modified organism (LMO) isolated fields. From the three types of rice fields, a total of 13,982, 14,105, and 12,730 individuals from the Bt-9, Ilmi-byeo and Dongjin-byeo were collected, respectively. Throughout the study, the analysis of variance indicated no significant differences (p < 0.05). Multivariate analysis showed that the abundance and diversity of plant dwelling insects were similar. The data on insect species population densities were subjected to principal component analysis (PCA), which did not distinguish among the three varieties, Bt-9 and the non-GM, reference cultivars, during the cultivation years. However, the results of the PCA analysis were completely divided into four groups based on the yearly survey areas. Therefore, there was no evidence for a negative impact of Bt-9 on the above-ground insects and spiders.

Scientific considerations for the biosafety of the off-target effects of gene editing in crops (유전자교정작물 내 비의도적 돌연변이의 안전성 논란에 관한 과학적 고찰)

  • Lee, Shin-Woo;Kim, Yun-Hee
    • Journal of Plant Biotechnology
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    • v.47 no.3
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    • pp.185-193
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    • 2020
  • The number of commercially approved gene-edited crops is gradually increasing, and in South Korea, it has led to intense investment in gene-edited crop development to increase international competitiveness. However, as with genetically modified crops, the safety of gene-edited crops regarding unexpected risks for humans and the environment is subject to an ongoing debate. In particular, unintentional "off-target effects" have become the center of controversy. In this review, we discuss typical plant characteristics (including somatic variation and ploidy), the extent of various off-target effects in genetically modified crops generated via horizontal transfer in nature, and the off-target effects in commercial genetically modified crops. We conclude that most off-target effects possibly occurring in gene-edited crops are not expected to be critically harmful to humans or the environment. Therefore, existing regulation for genetically modified crops should be enough for the risk assessment of gene-edited crops.

Comparative nutritional analysis for protopanaxadiol-enhanced genetically modified rice and its non-transgenic counterpart

  • Na Yeon Kim;Sung Dug Oh;Soo Yun Park;An Cheol Chang;Seong Kon Lee;Ye Jin Jang;So-Hyeon Baek;Yong Eui Choi;Jong-Chan Park;Doh Won Yun
    • Korean Journal of Agricultural Science
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    • v.51 no.2
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    • pp.239-249
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    • 2024
  • In the assessment of the biosafety of genetically modified (GM) crops, a comparative approach to identifying similarities and differences between transgenic and non-transgenic crops is helpful in identifying potential safety and nutritional issues. In this study, we aimed to compare the nutritional composition of a protopanaxadiol-enhanced genetically modified rice (PPD GM rice) with its non-transgenic counterpart. The nutritional profile of PPD GM rice was assessed against that of the parental rice cultivar 'Dongjin' to ascertain nutritional equivalence. No differences were observed between PPD GM and Non-GM rice cultivar in proximate analysis, mineral content, and amino acid composition. Although significant differences were observed in crude fat, crude protein, total dietary fiber, and some minerals between PPD GM rice and Dongjin, these variances fell within the range suggested by common cultivars (Anmi and Nipponbare) and Organization for Economic Cooperation and Development (OECD) data. Similarly, while some amino acids showed significant differences, these metabolites did not deviate from the OECD range. Principal component analysis (PCA) was conducted using the nutritional analysis data of PPD GM rice and Dongjin. The results revealed that PPD GM rice and Dongjin were grouped according to their respective cultivation years. This suggests that the variability in the nutritional composition of PPD GM rice tends to resemble that of the parental rice cultivar 'Dongjin' rather than being solely attributed to genetic modification. Overall, our findings indicate that the nutritional composition of PPD GM rice is substantially equivalent to that of its non-transgenic counterpart.