초록 ▼

□ 과제명 : GM작물의 환경위해성 평가
▶ 연구목적 :
○ GM 작물의 농업 환경위해성 평가 프로토콜 확립.
○ GM 작물의 농업 환경위해성 평가 심사 자료 생산
○ GM작물 실용화 체계 구축으로 국내 농업의 가치 향상 및 산업화 기반 구축
○ GM벼가 환경에 미치는 영향을 측정함으로써 GM벼가 환경에 미치는 영향을 최소화 하는 방향으로 재배방법의 표준화
▶ 주요연구내용 :
○ 1세부연구명 : 비타민A 강화 벼의 분자생물학적 특성구명
- 비타민A 강화벼의 분자생물학적 특성 구명 및 유전자

□ 과제명 : GM작물의 환경위해성 평가
▶ 연구목적 :
○ GM 작물의 농업 환경위해성 평가 프로토콜 확립.
○ GM 작물의 농업 환경위해성 평가 심사 자료 생산
○ GM작물 실용화 체계 구축으로 국내 농업의 가치 향상 및 산업화 기반 구축
○ GM벼가 환경에 미치는 영향을 측정함으로써 GM벼가 환경에 미치는 영향을 최소화 하는 방향으로 재배방법의 표준화
▶ 주요연구내용 :
○ 1세부연구명 : 비타민A 강화 벼의 분자생물학적 특성구명
- 비타민A 강화벼의 분자생물학적 특성 구명 및 유전자 안정성 분석
- 연차별 GM벼 및 베타카로틴 유전자 교잡계통에 도입된 유전자 8종에 대한 삽입양상 분석
- 비타민A 강화벼의 비표적 생물체 영향 평가
○ 2세부연구명 : 비타민A 강화 벼의 중, 남부 지역 농업적 특성 비교연구
- 지역별 재배학적, 농업적 특성 및 수량성 연차간 비교
- GM벼의 종자(교잡종) 특성 및 화분 특성 비교
○ 3세부연구명 : 비타민A 강화 벼의 병 발생 변화양상 비교연구
- 벼도열병 및 벼흰잎마름병에 대한 저항성 평가
- 연차간, 지역별 GMO 격리포장에서 자연발병 양상 평가
○ 4세부연구명 : 비타민A 강화 벼의 곤충 및 생물상 변화양상 비교
- 논포장내 곤충 및 거미상 비교
- 우점종 및 생물다양성 분석 및 대표 분류군별 밀도 및 동태 분석
○ 1협동연구명 : 비타민A 강화 벼의 대규모포장 농업적, 재배학적 특성 비교연구
- 비타민A 강화벼의 대규모 포장 농업적 특성 분석
- 비타민A 강화벼의 병 발생 변화양상 분석
○ 2협동연구명 : 비타민A 강화 벼의 유전자 이동성 및 잡초화 가능성 연구
- GM 벼의 포장 재배시 도입유전자이동/잡초화 최소화를 위한 재배 안전성 검정체계 및 표준 재배법 확립
- 비타민A 강화벼 품종의 화분특성 규명
- 안전성평가 프로토콜 작성을 위한 평가자료 생산
○ 5세부연구명 : 비타민A 강화 벼의 근권미생물 변화양상 비교연구
- 비타민A 강화벼와 비형질전환벼에 의한 논토양 우점미생물의 변화 비교 분석
- 분자생물학적 기법을 통한 비타민A 강화 벼에 의한 토착미생물상 변화 분석
- 형질전환 유전자의 토양미생물로의 수평적 이동 여부 조사
○ 6세부연구명 : 비타민A 강화 벼 도입 발현형질 후대 안정성 연구
- 비타민A 강화벼 도입 발현형질 후대 안전성 연구
- 비타민A 강화벼(일품여교배품종, BC3F4) 와 대조군인 낙동벼, 일품벼 포장/온실 재배.
비타민A 강화벼, 낙동벼(모본), 일품벼의 도입유전자들별 발현 안정성, 발현 패턴 분석(PCR 및 immu nostrip 분자생물학적방법)
○ 3협동연구명 : 여교배와 MAS를 이용한 안전화 계 선발 및 근동질 계통육성
- 베타카로틴 생성 근동질 벼 안정적 고정계통(Event)의 flanking region 해석
- 베타카로틴 생성 근동질 벼 안정적 고정계통(Event)의 증식 및 유전적 안정성과 농업적 형질 종합분석
○ 7세부연구명 : GM 콩의 장기적 생태영향 평가 연구
- 비의도적 환경방출 콩에 대한 환경위해요소 조사
- 제초제저항성 콩 환경방출과 관련한 야생환경내 GM 작물 생육양상 분석
○ 8세부연구명 : GM 콩의 토양미생물상 연구
- 제초제저항성 콩의 근권내 토양미생물상 영향 조사
- 토양미생물 수평이동평가요소 개발
○ 9세부연구명 : GM 채소 품종화를 위한 재배 및 안전관리 기술 개발
- 제초제저항성을 이용한 연차간 재배학적 및 농업적특성 비교 분석
- 수량 특성 및 환경변화 모니터링

Abstract ▼

The vitamin A enriched golden rice (PAC) was developed and confirmed to synthesis carotenoids (zeaxanthin, α-carotene and β-carotene) in rice endosperm. For biosafety assessment of golden rice, the stability of the insert genes in golden rice at genomic and protein expression level through different

The vitamin A enriched golden rice (PAC) was developed and confirmed to synthesis carotenoids (zeaxanthin, α-carotene and β-carotene) in rice endosperm. For biosafety assessment of golden rice, the stability of the insert genes in golden rice at genomic and protein expression level through different generations(T3~T7) was studied .Southern blot analysis confirmed that the multi-generation stability of the genes:PAC gene(CrtI and P sy), promoters(35S and Glb), terminators(NOS and P inI I ), selection marker(bar) and M AR gene. And it was also confirmed that the backbone DNA containing antibacterial gene(aadA) was not present in golden rice. Immuno-stripassay (PAT) showed bar gene was stably expressed through generations. These result will be filed to biosafety assessment document of Golden rice.
This study was conducted to investigate agronomic characteristics of vitamine A biofortified transgenic rice (VTR) compared to the wild type variety Nagdongbyeo(Wt). The innate characteristics of VTR, including leaf color, stem diameter and so on were mostly similar to those of Wt. The heading date of VTR was August 14 earlier two days than that of Wt in Suwon. The culm length of VTR was shorter than that of Wt, and the panicle length and panicle number of VTR was similar to those of Wt. However the some characteristics related to yield including ripening ratio and grain number per panicle in VTR were generally inferior to those of Wt. The average yield of milled rice of VTR was 378kg/10a lower 16.4% than that of Wt. The appearance of brown rice of VTR was worse than that of Wt to have so much white core. The percentage of brown rice recovery of VTR was lower 1.4% than that of Wt, and its protein and amylose contents were 6.2% and 20.1%, respectively.
To evaluate disease response of genetically modified(GM) rice cultivar to some important rice diseases was carried out for two years. GM rice cultivar increased Vitamin A contents showed similar diseases development patterns compared with original rice cultivar, Nagdongbyeo. In the inoculation test with blast isolates, Original cultivar showed susceptible reaction to all blast isolates but Vitamin A contents increased rice cultivar was showed resistant reaction against two races, 04-049 and R97-097. In the blast nursery test, original rice cultivar and GM rice cultivar were commonly showed susceptible reaction. 29 blast isolates collected from original rice cultivar were classified into 10 races and 30 blast isolates collected from GM rice cultivar were classified into 9 races. 7 races were isolated two rice cultivars for race classification. Seven races were commonly isolated in the two cultivars but three races, KI-301, KI-1113, KI-401, were only isolated from original rice cultivar and two races, KJ-105 and KI-317, were isolated from GM rice cultivar. These results indicated that diseases occurrence pattern of GM rice cultivar is very similar compared with original rice cultivar even though GM rice cultivar were showed resistant reaction against two blast isolates.
The structure and dynamics of arthropod (mainly insect and spider) community for ‘Nakdong’, and ‘Golden Rice’ (genetically modified vitamin A rice) were explored in small scale field trials during 2009, 2010, and 2011 using both powered suction device and sweep-net sampling methods. The taxa at family level found in Nakdong and Golden rice plots for three years consisted of 44-54 and 44-50 families, repectively. The many species which belonged to Aranea, Hemiptera, Homoptea, Coleoptera, Hymenoptera, and Diptera order appeared during the rice growing season. Hebivores and detritivores occupied about 90% of total individuals in both plots. The total number of taxa and population density were not significantly different between two experiment plots. The fauna in both plots was dominated by chironomids, planthoppers, rice weevils, aphids, and leafminers. The population density of dominant species was similar between two plots. The community diversity indices and cluster analysis also revealed that the arthropod community structure between Nakdong and Golden rice is very similar. Therefore, these results show that the cultivation of golden rice in field did not accompany a marked change in rice arthropd community.
This study was carried out to develope of macro-protocol and the biosafety guide ‘Vitamin A enforced rice’, modified vitamin A at large scale GMO field from 2009 to 2011. Commercialization of genetically modified (GM) plants will be required the assessment of risks associated with the release of GM plants that should include a detailed risk assessment of their impacts in the environment and human health. Prior to GM plant release, applicants should provide the information on GM crops for approval. A total of 4,700m2 GM　field, each of ‘Vitamin A enforced rice’ and ' Nakdong' were used for further analysis on agronomic traits and pathogenesis. Generally plant growth and panicle number in the transgenic lines were lower than in the wild type under field condition. The major agronomic traits such as plant growth, grain quality in ‘Vitamin A enforced rice’ were similar than in ' Nakdong' . The yield difference was due to reduced ripened rate and the 1,000 grain weight. The grain shape was similar donor plant, however, color of brown rice was different from segregation. Also the difference of pathogenesis reaction was not significant in the leaf blast and sheath blight between ‘Vitamin A enforced rice’ and ' Nakdong' . These results provided the agronomic data and pathogenesis for risk assessment analysis of ‘Vitamin A enforced rice’ and suggested that the macro-protocol could be useful to detect GM plants. The purpose of this study was to analyze the weediness potential associated with ‘Vitamin A enforced rice’ and to evaluate the relationships between growth characteristics and environmental adaption related to survival ability in the natural environment. ‘Vitamin A enforced rice’, developed from the donor variety ‘Nagdong’ and 3 varieties were used for this study. The ‘Vitamin A enforced rice’ were experimented according to ratio of ripened grain, shattering of grain, germination traits, germination ratio of low temperature, emergence ratio after wintering, ratio of viviparity, and growth ratio after ratooning ability. ‘Vitamin A enforced rice’ was not significant between ‘Nagdong’ and other varieties in seven characters. Germination ratio was similar than, however, germination ability of average was different from low temperature. Also, shattering of grain, germination ratio of alternating temperature was significant between ‘Nagdong’ and other varieties. 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 ‘Vitamin A enforced rice’, but natural enemy density on ‘Vitamin A enforced rice’ was a little higher. These results suggested that the relationships between growth characteristics and environmental adaption in ‘Vitamin A enforced rice’ could be applied to the stability of weediness potential and biosafety guide in GM rice.
Gene flow event from genetically modified (GM) rice to adjacent non-GM rice lines naturally happen in the field. Although GM rice lines containing desirable agronomic characteristics such as resistance to abiotic and biotic stresses can also be used to cross with non-GM cultivated rice and wild rice. This event can eventually lead to offsprings which possess traits that allows a better chance of survival in the field. However this phenomenon can have an enormous effect on the environment surrounding the rice field. Our first objective of studying gene flow is to determine the out-crossing rate of GM rice to cultivated rice. For the gene flow research, we used Hwangkembyeo (containing beta-carotene enhancing gene and bar gene) and Nakdongbyeo (mother plant of Hwangkembyeo). Results showed that most gene flow events occurred within the 1m range. In addition, most events happened within 30 cm and rare events were observed within 90cm range and beyond. The maximum distance for gene flow events was observed at 6m rangehowever the sharp cut point distance was on 60-90cm. The prevailing gene flow direction was North-West (NW) which coincided with the prevailing wind of South-eastern direction. Wind direction and distance were found to be the most important factors for determining rice out-crossing rate. But for more accurate prevention of GM gene flow, many factors should be considered like local weather condition, wind direction, speed and flowering time for safe cultivation.
The aim of this study was to investigate the possible impact of PAC rice on the soil microbial community. Microbial communities were isolated from the rhizosphere of PAC and Nakdong, parental variety and were subjected to be analyzed using both culture-dependent and molecular methods. The total counts of bacteria, fungi, and actinomycetes in the rhizosphere of transgenic and conventional rice were observed to have an insignificant difference. Denaturing gradient gel electrophoresis (DGGE) analysis of PCR-amplified 16S rRNA genes revealed that the bacterial community structures were very similar to each other and this genetic stability of microbial communities was maintained throughout the culture periods. Analysis of dominant isolates in the rhizosphere of transgenic and conventional rice showed that the dominant isolates from the soil of rice belonged to the Actinobacteria, H olophage, Clostridia and Betaproteobacteria. These results indicate that the PAC rice has no significant impact on the soil microbial communities. Beta-carotene producing transformants were produced in the background of "Nagdongbyeo", a Japonica rice cultivar. Introgression of the carotenoid locus in the transformant, PAC4-2 into the elite cultivar "Ilpumbyeo" was started. To initiate a backcrossing program, we surveyed 232 SSR markers and found that 38% of them were polymorphic between "Ilpumbyeo" as a recurrent parent and the PAC4-2 as a recipient parent. The selection strategy comprising foreground and background selection was employed. First, foreground selection was practiced in BC1,BC2, and BC3 generations using the transgene specific PCR-based marker in addition to visual scoring of the seed color. Marker-based background selection combined with phenotypic selection was employed from BC3F2 to BC3F4 generations. Blast search indicated that the transgene PAC4-2 was located between SSR markers, RM6 and RM482. 240 BC3F3 and 63 BC3F4 lines were evaluated for four agronomic traits including days to heading. Most of the lines were similar to Ilpumbyeo in agronomic traits evaluated. The percentage of PAC4-2 genome ranged from 4% to 21% with a mean of 12.5%, which was higher than the expected for an unselected BC3 backcross population. This could be explained by the fact that two genes for beta-carotene and the stripe virus resistance were targeted in this study. We selected 10 representative BC3F5 lines from 63 BC3F4 lines based on agronomic traits and carotenoids content. The selection strategy would be appropriate for the introgression of beta-carotene gene in a breeding program.
The aim of this study is to evaluate the gene flow from the genetically modified soybean to wild-type soybean (Glycin soja) plants in the field. We planted the glyphosate tolerant soybean (GM soybean) and planted wild type soybean plants as a pollen receptor around them. From the various distances from pollen source (GM soybean), we harvested 219,015 seeds by each row. We used cp4 epsps as a target gene. The cross-pollination frequency was evaluated by the survival rates the progeny plants after applying the glyphosate solution and then confirmed the presence of cp4 epsps gene by immunochemical chromatography and PCR analysis. Under the conditions of this experiment, pollen flow from GM soybean plants to wild-type soybean plants was extremely rare. 12 soybeans from 219,015 were determined to be hybrids between GM soybean and wild-type soybean. The frequency of gene flow was 0.0054% on average considering germination rate of wild-type soybean as 78%. The gene flow rates within 1.0m from the pollen source were 0.04% on average. From the studies, we can assume that the potential for gene flow to occur during soybean cultivation is assumed to be very low.
To investigate the effect of transgenic soybean on the soil microbial communities, an inbred line of transgenic soybean and four varieties of conventional soybean were employed for this experiment. The total counts of bacteria, fungi, and actinomycetes, denaturing gradient gel electrophoresis (DGGE), dominant members in the soil microbial community of transgenic soybean were compared with those of conventional soybean. Insignificant difference was observed in the colony forming unit from the rhizoplanes of transgenic and conventional soybean. Analysis using (DGGE) of PCR-amplified 16S rRNA genes revealed that the bacterial community structures were quite similar to each other in a given month, suggesting that there were no significant differences in bacterial communities between GM and non-GM soybean soils. Sequencing analysis using 16S rDNA PCR clones from the rhizosphere of transgenic and conventional soybeans revealed that the dominant class from the soil of transgenic soybean were the Arthrobacter, Gemmatimonas, and Niastella, while those from the soil of conventional soybean were Bacillus, M oorella, and P elobacter. When soil DNAs were analyzed with PCR using primers for the partial 35S and CTP, which was introduced into transgenic soybean, positive DNA bands were not found in GM and non-GM soybean soils. These results indicate that the transgenic soyeban has no significant impact on the soil microbial communities.
This study aimed to develop safety management and cultivation techniques for using genetically modified (GM) red pepper (Capsicum annum L.) resistant to herbicide as a new variety. Herbicide was applied to leaves of non-GM and GM Subicho and commercial varieties, Supermanita, 15, 30 and 45 days after planting (defined as 15-DAP, 30-DAP and 45-DAP treatment). In result, both of Supermanita and non-GM Subico plants were dead at the 15-DAP treatment, whereas GM Subicho survived approximately 90 percent at the treatment. Fruit yield was the highest at the 45-DAP treatment regardless of varieties, approximately 153 g per plant, than other treatments, while there was no treatment effect on weight, length and width of fruit. At the 45-DAP treatment, Supermanita had higher weight, length and width of fruit than other varieties, while there was no difference in the parameters between non-GM and GM Subicho. Anthracnose infection rate on fruits was the highest at the 30-DAP treatment than other treatments, regardless of tested varieties. Asta value of GM Subicho was higher at the 15-DAP treatment, approximately 233, than other treatments. At the 30-DAP treatment, no variety variation in the asta value was found, while both non-GM and GM Subicho had higher asta values than Supermanita at the 45-DAP treatment.

목차 Contents

• 제출문 ... 1
• 목차 ... 2
• 요약문 ... 4
• I. 제목 ... 4
• II. 연구개발의 목적 및 필요성 ... 4
• III. 연구개발의 내용 및 범위 ... 6
• IV. 연구개발결과 ... 7
• V. 연구개발결과의 활용계획 ... 12
• SUMMARY ... 13
• 제1장 서론 ... 19
• 제2장 국내외 기술개발 현황 ... 22
• 제1절. 국내 연구 현황 ... 22
• 제2절. 국외 연구 현황 ... 23
• 제3장 연구개발수행 내용 및 결과 ... 25
• 제1절. 비타민 A 강화벼의 분자생물학적 특성 구명 및 후대 안전성 연구 ... 25
• 제2절. 비타민 A 강화벼의 중,남부 지역 농업적 특성 비교연구 ... 45
• 제3절. 비타민 A 강화벼의 병 발생 변화양상 비교연구 ... 51
• 제4절. 비타민 A 강화벼의 곤충 및 생물상 변화양상 비교 ... 54
• 제5절. 비타민 A 강화벼의 대규모포장 농업적, 재배학적 특성 비교 연구 ... 64
• 제6절. 비타민 A 강화벼의 유전자 이동성 및 잡초화 가능성 연구 ... 97
• 제7절. 비타민 A 강화벼의 근권미생물 변화양상 비교연구 ... 120
• 제8절. 여교배와 MAS를 이용한 안전화 계통 선발 및 근동질 계통 육성 ... 133
• 제9절. GM 콩의 장기적 생태영향 평가 연구 ... 140
• 제10절. GM 콩의 토양미생물상 연구 ... 147
• 제11절. GM 채소 품종화를 위한 재배 및 안전관리 기술 개발 ... 159
• 제4장 연구개발목표 달성도 및 대외기여도 ... 164
• 제1절 목표대비 달성도 ... 164
• 제2절 정량적 성과 ... 168
• 제5장 연구개발결과의 활용계획 ... 173
• 제6장 중요 변동사항 ... 174
• 제7장 참고문헌 ... 176
• 주요 결과 요약서 ... 189