Assessment of environmental impact of vitamin A-enhanced soybeans and hybrid soybeans

오성덕,최지은,장예진,이성곤,이성곤,장안철,윤도원 충남대학교 농업과학연구소 2023 Korean Journal of Agricultural Science Vol.50 No.4

An understanding of safety problems pursuant to environmental release of GM (Genetically Modified) crops is considered important. Among the recognized safety problems, the possibilities of weediness and ecosystem invasion are constantly being validated. We herein compared the growth characteristics and germination rate of soybeans formed by hybridization with vitamin A-enhanced soybeans carrying an introduced gene that increases β-carotene content. We also examined overwintering, survival, and weed competitiveness to evaluate hybrid ecological impact on long-term unmanaged cultivatable land. These studies revealed that the hybrid soybeans exhibited intermediate growth characteristics and germination rate compared with the vitamin A-enhanced soybeans and wild soybeans, or exhibited traits similar to those of the maternal strain. Overwintering experiments were conducted by planting seeds at depths of 0, 5, 10, and 20 cm and recovering them after three or five months. After five months, all seeds at depths more than 5 cm lost viability. Among seeds recovered after three months, only wild soybeans retained viability at depths of more than 5 cm. Survival and weed competitiveness were assessed by sowing each type of seed and performing no irrigation, or pest or weed control. Quantitative assessment of numbers of individual soybean plants that appeared in the experimental plot revealed that all plants germinated after sowing, but only wild type plants survived overwintering. These studies suggest that both GM soybeans and hybrid soybeans cannot survive in uncultivated land even if they are released into the environment, which indicates less possibility of ecosystem invasion and weediness.

Impact of vitamin-A-enhanced transgenic soybeans on above-ground non-target arthropods in Korea

오성덕,하기훈,박수윤,이성곤,윤도원,이기종,서상재 충남대학교 농업과학연구소 2021 Korean Journal of Agricultural Science Vol.48 No.4

In order to confirm the safety of a genetically modified organism (GMO), we assess its potential toxicity on non-target insects and spiders. In this study, the effects of GM soybean, a type of vitamin-A-enhanced transgenic soybean with tolerance to the herbicide glufosinate, were assessed under a field condition. The study compared this vitamin-A-enhanced transgenic soybean and a non-GM soybean (Gwangan) in a living modified organism (LMO) isolated field of Kyungpook National University (Gunwi) and the National Institute Agricultural Sciences (Jeonju) in the Republic of Korea in 2019 - 2020. In total, 207,760 individual insects and arachnids, representing 81 families and 13 orders, were collected during the study. From the two types of soybean fields, corresponding totals of 105,765 and 101,995 individuals from the vitamin-A-enhanced transgenic soybean and Gwangan samples areas were collected. An analysis of variance indicated no significant differences (p < 0.05). A multivariate analysis showed that the dominance and richness outcomes of plant-dwelling insects were similar. The data on insect species population densities were subjected to a principal component analysis (PCA) and an orthogonal partial least squares-discriminant analysis (OPLS-DA), which did not distinguish between the two varieties, i.e., the vitamin-A-enhanced transgenic soybean and the non-GM soybean in any cultivated field. However, the results of the PCA analysis could be divided overall into four groups based on the yearly survey areas. Therefore, there was no evidence for the different impact of vitamin A-enhanced transgenic soybean on the above-ground insects and spiders compared to non-GM soybean.

유전자 변형 비타민E 강화콩의 환경 영향 평가 : 비타민E 강화콩의 급이가 물벼룩(Daphnia magna)에 미치는 영향

오성덕,윤도원,손수인,이범규,이기종,장안철 한국국제농업개발학회 2016 韓國國際農業開發學會誌 Vol.28 No.2

The vitamin E enhanced transgenic soybean was developed by introducing a perilla γ- tocopherol methyltransferase gene (γ-TMT) under the control of pea vicilin promoter and a selection marker, phosphinothricin acetyltransferase (PAT) gene. With regard to the potential problems of safety, the non-target organism evaluation is required as an essential element for the environmental risk assessment of genetically modified (GM) crops. We studied the effects of the vitamin E enhanced transgenic soybean feeding on survival of Daphnia magna which is commonly used as a model organism in ecotoxicological studies. The Daphnia magna was fed on vitamin E enhanced transgenic soybean and non-genetically modified (non-GM) soybean (Willams 82) at 0, 1,000, 1,800, 3,240, 5,830, 10,500 and 20,000 mg/L concentrations, respectively. The GM soybean used for the test was confirmed to have the γ-TMT/PAT gene expression by the enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR) analysis. The results showed that there was no significant differences between cumulative immobilities of Daphnia magna fed on GM soybean and non-GM soybean. The 48hr-EC50 values showed no significant differences between GM soybean (2,416 mg/L) and non-GM soybean (2,408 mg/L). The results of this study suggested that there was no significant differences in toxicity for Daphnia magna between GM soybean and non-GM counterpart.

가뭄저항성벼(Agb0103)의 분자생물학적 특성과 안전성 평가

오성덕,이기종,원소윤,손수인,이시명,박순기,류태훈 한국육종학회 2014 한국육종학회지 Vol.46 No.4

가뭄저항성벼의 복수세대에 대한 후대안정성을 서던 분석과 PCR로 분석한 결과, 가뭄저항성벼의 Agb0103 T4~T6 세대에서는 도입된 모든 유전자들이 안정적으로 도입되어 있으며, T-DNA 구성요소 이외의 backbone DNA는 가뭄저항성벼에 삽입되지 않았음을 확인하였다. 목적 유전자인 CaMsrB2와 제초제 저항성 선발 마커인 bar 유전자가 가뭄저항성벼 Agb0103의 T4~T6 세대에서 안정적으로 발현됨을 검증하였다. 제초제 저항성 선발 마커로 도입된 PAT 단백질의 발현 분석 결과에서도 Agb0103 T4~T6의 3세대에서 생육시기별, 부위별로 안정적으로 발현됨을 입증하였다. 도입유전자의 삽입 위치를 확인한 결과, 가뭄저항성벼 Agb0103의 도입유전자가 벼 8번 염색체 내에서 intergenic한 상태로 안정적으로 유지되고 있음을 확인하였다. 이상의 분석 기법을 통해 복수세대에서 가뭄저항성벼의 도입 유전자들이 안정적으로 유지되고 목적 단백질들이 안정적으로 발현되고 있음을 확인하였다. Genetically modified (GM) crops have been developed worldwide through the recombinant DNA technology and commercialized by various agricultural biotechnology companies. Commercialization of GM crops will be required the assessment of risk associated with the release of GM crops. The purpose of this research is a molecular characterization of introduced T-DNA in transgenic rice T4∼T6 generation lines harboring a pepper MsrB2 gene under the control of stress inducible Rab21 promoter, as a part of biosafety evaluation for drought-tolerant transgenic rice (Agb0103). We identified the structure and sequence of transformation vector of T-DNA and analyzed insertion sites, flanking sequences, and generational stability of inserted T-DNA in transgenic rice lines. The transformation vector was consisted of right border, a drought-tolerant CaMsrB2 gene unit (Rab21 promoter:CaMsrB2:PinII terminator), a selectable marker herbicide resistance unit (CaMV 35S promoter:bar:Nos terminator), and left border in sequential order. Based on the adaptor-ligation PCR and whole genome sequence database, we confirmed that T-DNA was introduced 2 copies (head to head type) at the position of 2,471,957∼2,472,049 bp of chromosome No. 8. From the generational stability study, T-DNAs were stably inherited through the T4 to T6 generations, and also stable expression of bar gene from T-DNA was confirmed. It was also confirmed that the backbone DNA of transformation vector containing antibacterial gene was not present in Agb0103 rice genome. These results will be filed to biosafety assessment document of Agb0103.

비타민A 강화벼의 환경위해성 평가 : 4세대의 후대안정성 연구

오성덕,박수윤,손수인,김재광,박종석,박순기,조현석,안병옥,이기종 한국육종학회 2012 한국육종학회지 Vol.44 No.4

비타민A 강화벼의 복수세대에 대한 후대안정성을 Southern blot과 PCR로 분석한 결과, 비타민A 강화벼의 PAC T3~T6 세대에서는 도입된 모든 유전자들이 안정적으로 도입되어 있으며, backbone DNA는 비타민A 강화벼에 삽입되지 않았음을 확인하였다. 선발 마커로 도입된 PAT 단백질의 발현 분석 결과에서도 PAC T3~T6 복수세대에서 생육시기별 부위별로 안정적으로 발현됨을 입증하였으며, 최종 목적 산물인 카로티노이드 분석 결과에서도 모품종인 낙동벼에 비해 비타민A 강화벼에서 β-carotene은 10.6배 함량이 증가되고, zeaxanthin과 α-carotene는 생성되었음을 확인하였다. 이상의 분석기법을 통해 복수세대에서 비타민A 강화벼의 도입 유전자들이 안정적으로 유지되고 목적 단백질들이 안정적으로 발현되고 있음을 확인하였다. The carotenoid-biofortified (PAC) rice was developed and confirmed to synthesis carotenoids (zeaxanthin, α-carotene and β-carotene) in rice endosperm. For biosafety assessment of PAC rice, the stability of the insert genes in carotenoid- biofortified rice at genomic and protein expression level through different generations (T3~T6) was studied. Southern blot analysis confirmed that the multi-generation stability of the genes : PAC gene (CrtI and Psy), promoters (35S and Glb), terminators (NOS and PinII), selectable marker (bar) and MAR gene. It was also confirmed that the backbone DNA containing antibacterial gene (aadA) was not present in PAC rice. Immuno-strip assay (PAT) showed that the bar gene was stably expressed through generations. HPLC analysis confirmed that carotenoids were consistently detected through T3~T6 generations. These results will be filed to biosafety assessment document of PAC rice.

「연강첩장도(烟江疊嶂圖)」에 덧붙여진 왕선과 소식의 제화시 연구

오성덕,무금연 대한중국학회 2019 중국학 Vol.66 No.-

During the Song Dynasty, the Wang An-Seok’s New Law brought about political instability, and many literary men went through the literary dark period, which was disciplined for opposing the new law. However, in spite of this persecution, the Song dynasty has become the prime time of literary and landscape painting development unique to the history of China. This unique art world of china, as named with Painting & Letter Poetry, incorporates the artistic aestheticism of poetry and painting into one single work with its implicitly conceals the ideas and philosophy of the literary creator in the frame of his work. These works of art are melted in poetry and paintings, with metaphorical implications that are different from those of Western artistic works. Especially, the metaphorical implication of landscape painting and literary painting is the product of the era of sharing the metaphorical meanings hidden in the works, as the monks refrained from using words and phrases to avoid persecution like letter jade (文字獄). Paintings and Poetry, which are different field of art world on the different spaces, became the Painting & Letter Poetry of China's unique art world and different spaces are expanding their artistic value in a single space. I hope that it will be a chance to re-think about the value and the meaning of literary history of the Painting & Letter Poetry through the study of Su Shi and Wang Shen’s poetry that have been added to the landscape painting of Yan Jiang Die zhang Map (烟江疊嶂圖), which is the representative in the Song Dynasty of China. 송대 신종때는 왕안석 신법 주창으로 정치 불안이 야기되었고, 수많은 문인들은 신법에 반대한다는 이유로 징계를 받아 폄적되는 문학적 암흑기를 거치기도 했다. 하지만, 이런 핍박 속에서도 송나라는 서양의 작품세계와는 차별화된 중국 고유의 문인화, 산수화를 꽃피우기에 이른다. 이 독특한 동양의 작품세계는 시와 그림이 지향하는 작품적 심미주의를 하나의 작품 속에 통합하고, 작품이라는 프레임 속에 문인들의 사상과 철학이 암시적으로 숨겨진다. 이러한 작품세계는 사실을 사실적으로 드러내고 분석하려는 서양의 작품세계와는 차별화된 함축적 비유나 은유적 함의가 시와 그림 속에 그대로 녹여져 있다. 특히, 산수화나 문인화의 은유적 함의는 문자옥(文字狱)과 같은 박해를 피하기 위해 선비들이 언행을 삼가고, 마음을 그림 속에 투영하게 되면서, 작품 속에 숨겨진 은유적 의미들을 서로 공유하는 시대풍조의 산물이 되었다. 그림과 시라는 서로 다른 공간의 예술 활동이 중국 특유의 제화시라는 작품으로 서로 다른 공간들이 하나의 공간에서 그 예술적 가치를 더없이 확대하고 있는 것이다. 중국의 제화시하면 떠오르는 송대 대표적인 산수화 「연강첩장도(烟江疊嶂圖)」에 덧붙여진 왕선과 소식의 제화시 연구를 통해 제화시의 가치와 문학사적 의미를 다시 생각해 보는 계기가 되었으면 한다.

연구보문 : 자연과학 ; 비타민A 강화 벼 분자생물학적 분석 및 물벼룩 급이 효과

오성덕,이기종,박수윤,손수인,류태훈,김재광,김진서,안홍일,하선화,박종석,안병옥,조현석,서상재 韓國國際農業開發學會 2012 韓國國際農業開發學會誌 Vol.24 No.4

A carotenoid-biofortified (PAC) rice was generated by inserting phytoene synthase (Psy) and carotene desaturase (CrtI) genes isolated from Capsicum annuum cv. Nockwang and Pantoea ananatis into the genome of a conventional variety of rice (Nakdongbyeo). For biosafety assessment, we evaluated the effects on survival of Daphnia magna which is a commonly used as a model organism in ecotoxicological studies. D. magna fed on PAC rice and non-GM rice (Nakdong) grown in the same environment (100% ground rice suspension). The PAC rice was confirmed to have the insertion of T-DNA and protein expression by the Southern blot and HPLC analysis. Feeding study showed similar cumulative immobility and abnormal response of Daphnia magna between PAC rice and non-GM counterparts. 48hr-EC50 values of PAC rice and non-GM rice showed 3,311 mg/L (95% confidence limits: 2,901.39 - 3,779.23 mg/L) and 3,655 mg/L (95% confidence limits: 3,156.71 - 4,232.86 mg/L) respectively, but there were not statistically significant.

대규모 GMO 포장에서 가뭄저항성벼(Agb0103)가 곤충다양성에 미치는 영향

오성덕,김진서,이기종,류태훈,서상재 한국육종학회 2014 한국육종학회지 Vol.46 No.2

This study was carried out to develop of macro-protocol and the biosafety guide for drought-tolerant transgenic rice (Agb0103) at large scale GMO field, a total of 4,700 m2. In GMO quarantine area of Kyungpook National University, insect species diversities and population densities on Agb0103 and wild type (Ilmi) were investigated. There was no difference between the population densities of insect pests and natural enemies on two varieties, while sometimes insect pest density on Ilmi was slightly higher than on Agb0103, but natural enemy density on Agb0103 was a little higher. These results provided the insect diversity for risk assessment analysis of Agb0103 and suggested that the macro-protocol could be useful to detect GM plants. 본 연구는 대규모 GMO 격리포장에서 고추 유래 가뭄저항성CaMsrB2유전자가 도입된 가뭄저항성벼(Agb0103)의 안정성 평가에 대한 가이드라인 및 프로토콜을 개발하고자 수행하였다. 경북대학교 농업생명과학대학 GMO 실습격리포장에서 총 사용면적 4,700 m²에서 Agb0103와 모품종인 일미벼에 대해 거미류를 포함한 곤충류 다양성을 조사하였다. 조사기간 동안 채집된 개체들은 기능별로 해충군, 천적군, 거미군으로 크게 구분하여 계수하였으며, 조사된 개체군의 해충군, 천적군, 거미류 등의 밀도는 기능군 별로 Agb0103와 일미벼간에 유의성을 보이지 않았다. 해충의 발생양상을 비교 분석한 결과, 전반적으로 일미벼에서 발생량이 다소 많은 경향으로 나타났으나, 대부분의 시기별 발생량 평균 간의 차이는 없는 것으로 나타나서 Agb0103와 일미벼과의 해충 발생양상에는 유의성을 찾기가 어렵다고 판단된다. 또한 천적류 발생양상은 해충 발생양상과는 달리 Agb0103에서 발생량이 다소 많은 경향으로 나타났으나, 시기별 평균 간 차이는 없는 것으로 분석되어 천적 발생양상에도 특이한 유의성을 없다고 판단된다. 따라서 가뭄저항성 CaMsrB2유전자가 도입된 Agb0103에서의 해충 및 천적의 개체군 밀도 및 발생양상은 모본인 일미벼에서와 차이가 없는 것으로 조사되었다.

LMO격리포장에서 비타민A강화콩 재배가 곤충다양성에 미치는 영향

오성덕,박수윤,이성곤,윤도원,이강섭,서상재 한국육종학회 2020 한국육종학회지 Vol.52 No.4

In this study, conducted in living modified organism (LMO) isolation fields, we sought to develop environmental risk assessmentprocedures for identifying the potential effects on non-target above-ground insects and spiders within agroecosystems cultivated with vitaminA-enhanced transgenic soybean with tolerance to the herbicide glufosinate. To this end, we investigated insect/arachnid species diversitiesand population densities on vitamin A-enhanced transgenic soybean and non-GM soybean (Gwangan) grown in LMO quarantine areas ofKyungpook National University (Gunwi) and the National Institute of Agricultural Sciences (Jeonju). In total, 93,419 individual insects andarachnids, representing 65 families in 12 orders, were captured during the study. In Gunwi, totals of 17,110 and 17,627 individual insectsand arachnids were collected from vitamin A-enhanced transgenic soybean and Gwangan, respectively, whereas in Jeonju, totals of 28,621and 30,061 individuals were collected from vitamin A-enhanced transgenic soybean and Gwangan, respectively. Although we detected nosignificant differences among the population densities of insect pests, natural enemies, and other insects on vitamin A-enhanced transgenicsoybean and Gwangan grown within the same field, the population densities of these insects were found to be higher in Jeonju than thosein Gunwi. Throughout the study, analysis of variance indicated no significant differences (p<0.05) in insect/arachnid populations, and multivariateanalysis indicated that the abundance and diversity of plant-dwelling insects were similar within the same fields.

베타카로틴 강화 GM콩과 야생콩간 교잡콩의 주요 영양성분 비교분석

오성덕,장예진,이경민,이기종,서상재,류태훈,박수윤 한국국제농업개발학회 2020 韓國國際農業開發學會誌 Vol.32 No.4

1. 본 연구는 GM콩이 비의도적으로 자연에 방출되었을 시 야생콩과의 교잡에 의해 발생 가능한 교잡콩을 모니터링하고 영양학적 측면에서 안전성을 평가하기 위하여 수행하였다. 이를 위해, 베타카로틴 강화 GM콩과 야생콩, 두 품종간 인위적으로 생산한 교잡콩, GM콩의 모본이자 일반 재배 품종인 광안콩 등 4품종을 국내 2개 지역에서 재배하고 콩 종자를 대상으로 주요 영양성분 38종을 분석하였다. 2. 재배 환경에 영향을 받지 않고, GM콩 및 야생콩과 비교해 교잡콩에서 유의적 차이를 보이는 성분은 조단백질, 조섬유, alanine, glycine, leucine, serine, tryptophan, oleic acid, linolenic acid, arachidonic acid 등 10종이었다. 하지만, 분석한 모든 시료의 성분은 국내외 자연 범위에 포함되는 수준이었다. 3. PCA분석 결과, 주요 영양성분 함량 변이는 GM콩은 모본과 유사하였고, 교잡콩은 GM콩 보다는 야생콩과 유사하였다. 즉, 분석한 4품종의 영양성분의 변이는 재배 환경보다는 유전적 요인에 의해 더 크게 영향을 받음을 알았다. 4. 본 연구의 결과는 GM콩과 야생콩간 교잡콩의 영양학적 안전성을 분석한 최초의 보고이며, 이는 향후 GM콩의 비의도적 자연 방출을 모니터링하는데 있어 유용한 기초 자료로써 활용 가치가 있을 것이다. This study was conducted to evaluate the nutritional safety and monitor hybrid soybean that can occur due to hybridization between wild soybean and genetically modified (GM) soybean when GM soybean is unintentionally released to environment. For this, β-carotene-enhanced GM soybean, wild soybean, artificially produced hybrid soybean between the two varieties, and ‘Gwangan’, the parent cultivar of GM, are cultivated in two areas in Korea and 38 nutrients were analyzed from the seeds of four varieties. The nutrients that showed significant differences in hybrid soybeans compared to GM and wild soybeans which were not affected by the cultivation environment were crude protein, crude fiber, alanine, glycine, leucine, serine, tryptophan, oleic acid, linolenic acid, and arachidonic acid. However, the levels of all nutrients obtained from samples analyzed were within the natural variation. As a result of PCA analysis, it was found that the variation of nutrient composition of the four varieties analyzed was more affected by genetic factors than by environmental factors such as growing conditions. Taken together, the nutritional safety of hybrid soybean is not different from cultivated soybean and wild soybean, and ten nutrients showing significant differences are possible as important components for identifying and monitoring hybridized soybeans. To our knowledge, this is the first study to assess the nutritional safety of hybrid soybean developed by hybridization between GM and wild soybeans, and the results of this study will be valuable as basic data for monitoring GM soybean when the soybean is unintentionally released to environment.