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 done to develop environmental risk assessments and a biosafety guide for insect-resistant genetically modified rice at a LMO (Living Modified Organism) isolation field. In the LMO quarantine area of Kyungpook National University, the species diversities and population densities of non-target insects found on insect-resistant genetically modified rice (Bt-T) resistant to Cnaphalocrocis medinalis and on non-GM rices (Dongjin-byeo and Ilmi-byeo) were investigated. The Bt-T event was therefore evaluated under field conditions to detect possible impacts on the above ground insects and spiders. The study compared transgenic rice and two non-GM reference rices, Dongjinbyeo and Ilmi-byeo, at Gunwi in Southern Korea in 2016 and 2017. A total of 9,552 individuals from 51 families and 11 orders were collected from the LMO isolation field. In the three types of rice fields, a total of 3,042 individuals from the Bt-T, 3,213 individuals from the Dongjin-byeo, and 3,297 individuals from the Ilmi-byeo were collected, respectively. There was no difference between the population densities of the non-target insect pests, natural enemies and other insects on the Bt-T and non-GM rices. The data of insect species population densities were subjected to principal component analysis (PCA), which not distinguished among the three varieties, GM and non-GM, reference cultivar in all cultivation years. However, PCA results were clearly separated the samples based on the cultivation years. These results suggest that insect species diversities and population densities during plants cultivation are determined by environmental factors (growing condition and seasons) rather than by genetic factors.

Insect-resistant transgenic (Bt-9) rice was generated by inserting the mCry1Ac1 a modified gene from the soil bacterium Bacillus thuringiensis into the genome of a conventional variety of rice (Ilmibyeo). 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. For biosafety assessment, we studied the effects on survival of cantor Daphnia magna, a commonly used as a model organism in ecotoxicological studies. D. magna fed on Bt-transgenic rice (Bt-9) and its near non-GM counterparts (Ilmibyeo) grown in the same environment (100% ground rice suspension). The Bt-9 rice was comfirmed to have the insertion of T-DNA and protein expression by the PCR and ELISA analysis. Feeding study showed similar cumulative immobility and abnormal response of Daphnia magna between Bt-9 rice and Ilmibyeo. 48hr-EC50 values of Bt-9 and Ilmi rice showed 4,400 mg/L (95% confidence limits: 3861.01 ~ 5015.01 mg/L) and 5,564mg/L (95% confidence limits: 4780.03 ~ 6476.93 mg/L) respectively. The rice NOEC (No observed effect concentration) value for D. magna was suggested 1,620mg/L. We conclude that the tested Bt-9 and Ilmi similar cumulative immobility for D. magna the widely used model organism. We found out that there is strong possibility that the growth of Bt-9 didn’t affect to non-target insects.

There are studies on the assessment of non-edible transgenic plants on soil microbial communities. In this research we evaluated the effect of virus-resistant trigonal cactus on soil microbial communities of the rhizosphere. Soil samples are collected and compared in genetically modified (GM) and non-GM trigonal cactus cultivation fields during vegetative growth period and post-harvest period. Biolog EcoplateTM was used to evaluate the functional diversity of soil microbial communities. There were no significant differences between the GM and non-GM soil samples collected during the vegetative growth period. However, we observed temporary difference in carbon substrate utilization. Principal component analysis showed that soil microbiota was influenced not by presence of GM or non-GM trigonal cacti, but rather by the cultivation period. Denaturing gradient gel electrophoresis fingerprinting revealed that virus-resistant trigonal cactus cultivation had insignificant effect on soil microbial communities including dominant rhizosphere bacteria, actinomycetes, and fungi. We found no clear evidence of GM trigonal cactus cultivation affecting the functional diversity of soil microbial communities.

Rice is the most important staple food and the consumption is increasing in Europe and America. It is expected that the rice price will be increased due to the unstable supply and demand because of abiotic stress. Experts expect that the rice supply of Korea will be deficient in 2050 for the reason of global warming. Conventional breeding methods need to be supplemented with recent biotechnology to meet the problem such as growing population and climate change. Using rice biotechnology, a number of agronomically important traits and nutritional value have been improved. In this paper, the current situation of rice cultivation and production situation and supply and consumption in the major country were investigated to prepare future food security. Also the research trends of rice biotechnology and kinds of transformed rice as well as safety issues were reviewed by considering the scientific, social and economic values of rice.

비타민A 강화벼와 ‘낙동’의 미꾸리(Misgurnus anguillicaudatus)와 잉어(Cyprinus carpio)에 대한 급성독성시험을 실시한 결과48시간 및 96시간-LC50은 1,000mg/L이상으로 나타났다. 48시간 및 96시간 무영향농도(NOEC)는 1,000mg/L이었다. 급성독성 시험기간 중 비타민A 강화벼와 낙동벼간의 pH, DO, 수온,체중 및 전장에 대한 유의적인 결과는 나타나지 않았다.

물벼룩 급성 독성 평가를 위한 비타민A 강화벼의 분자생물학적 특성을 분석한 결과, Southern blot에서 베타-카로틴 생합성을 위한 Psy와 CrtI 유전자들이 one-copy로 도입됨을 확인하였으며, 선발마커인 Bar 유전자의 단백질 검출 immunostrip 분석에서도 비타민A 강화벼에서만 검출되었다. 비타민A 강화벼의 목적하는 최종 산물인 베타-카로틴 함량도 낙동벼에 비해 8.9배 증가됨을 확인하였다. 비타민A 강화벼와 낙동벼의 농업환경 생물지표종인 물벼룩(Daphniamagna)에 대한 급성독성시험을 실시한 결과, 비타민A 강화벼의 48시간-EC50은 3,311.40 mg/L(95% 신뢰한계 : 2,901.39 ~ 3,779.23 mg/L), 무영 향농도(NOEC)는 1,800 mg/L였고, 낙동벼는 48시간-EC50은 3,655.23 mg/L(95% 신뢰한계 : 3,156.71 ~ 4,232.86 mg/L), 무영향농도는 1,800 mg/L였다. 따라서 Psy와 CrtI 유전자가 형질전환된 비타민A 강화벼 및 낙동벼가 환경 지표생물종인 물벼룩에 미치는 영향 평가 결과 상대적 동등성을 보였으며, 이는 Psy와 CrtI 유전자의 단백질 노출이 물벼룩에 부정적인 영향을 미치지 않은 것으로 판단된다.

This study was carried out to develop of environmental risk assessments and the biosafety guide for Vitamin E enhanced transgenic soybean at LMO (Living Modified Organism) isolation field. In LMO quarantine area of National Institute of Agricultural Sciences, insect species diversities and population densities on vitamin E enhanced transgenic soybean and non-GM soybeans (Willams 82 and Seoritae) were investigated. A total of 17,717 individuals of 77 species from 8 orders were collected in LMO isolation field. In three type soybeans field, total of 5,250 individuals in Vitamin E enhanced transgenic soybean, 5,510 individuals in Willams 82, and 6,957 individuals in Seoritae were collected, respectively. There was no difference between the population densities of insect pests, natural enemies and other insects on Vitamin E enhanced transgenic soybean and Willams 82, while natural enemies density on Seoritae was higher than on Vitamin E enhanced transgenic soybean, but insect pests density on Vitamin E enhanced transgenic soybean was higher. These results provided the insects diversity for risk assessment survey of Vitamin E enhanced transgenic soybean and suggested that the guideline could be useful to detect LMO crops.

Lactic acid bacteria (LAB) are industrially important microorganisms for probiotics. The recent widespread application of LAB for preparation of functional food is attributable to the accumulating scientific evidence showing their beneficial effects on human health. In this study, we isolated and characterized plant-derived LAB that show angiotensin-converting enzyme (ACE) inhibitory and antioxidant activities. The selected strain K2 was isolated from Kimchi, and identified as Lactobacillus plantarum by 16S rRNA gene analysis. The strain grew under static and shaking culture systems. They were also able to grow in different culture conditions like 25℃∼37℃ temperature, 4∼10 pH range and ∼6% NaCl concentration. L. plantarum K2 was highly resistant to acid stress; survival rate of the strain at pH 2.5 and 3 were 80% and 91.6%, respectively. The strain K2 also showed high bile resistance to 0.3% bile bovine and 0.3% bile extract with more than 74% of survival rate. The cell grown on MRS agar plate containing bile extract formed opaque precipitate zones around the colonies, indicating they have bile salt hydrolase activity. The strain showed an inhibitory activity against pathogenic bacteria such as Escherichia coli, Staphylococcus aureus and Listeria monocytogenes; antibacterial activity was probably due to the lactic acid. The K2 strain showed relatively higher autoaggregation values, antihypertensive and antioxidant activities. These results suggest that L. plantarum K2 could be not only applied as a pharmabiotic for human health but also is also starter culture applicable to fermentative products.

본 연구에서는 제초제 저항성인 Ab벼와 해충저항성 Bt벼, 이들의 모본인 동진벼의 현미에서 일반성분, 아미노산, 무기물, 지방산, 비타민 및 항영양소를 분석하여 유전자변형 벼(Ab, Bt)의 영양성분 조성 차이를 비교하고자 수행하였다. 그 결과 무기물의 철분이 모본인 동진벼와 유전자 변형 벼(Ab, Bt) 사이에 약 2배의 차이를 나타내었지만 codex 범위 내에 존재하였고, 비타민 류에서는 Bt벼와 동진벼 간의 유의적인 차이가 나타나지 않은 반면 Ab벼의 비타민 B1, B7, E에서 유의적인 차이를 보였다. 또한 트립신 저해제는 시료 모두에서 0.1 TIU/mg 미만으로 극소량 검출되었다. 총 46가지 분석 성분 중 17가지가 함량 및 조성에 유의적인 차이를 보였으나 이러한 차이는 이들의 모본인 동진벼, OECD 표준기술서, 그리고 일반 상업화 품종의 범위 내에 존재해 영양학적 측면에서 실질적으로 차이가 없는 것으로 판단된다.

Styela clava tunic is generated in large amounts as a waste from S. clava processing plants and leads to environmental problems. It destroys the beach scenery and causes a bad smell and pollution by trashing on the seashore. Therefore, purpose of this study was to investigate antioxidant and antihypertensive activities of different solvent extracts from S. clava tunic for recycling of fishery waste. Antioxidant and antihypertensive activities of all extracts were concentration-dependent. Of extracts, hot water extract showed the highest DPPH radical scavenging activity with the lowest effective concentration (EC50) value (0.733 mg/ml). Chloroform extract exhibited the highest metal chelation activity with the lowest EC50 value (2.696 mg/ml). Autoclaved water extract showed the highest NO radical scavenging activity with the lowest EC50 value (0.491 mg/ml) and n-hexane extract showed the highest reducing power (A700=1.897 at 100 mg/ml). And n-butanol extract showed the highest SOD-like activity with the lowest EC50 value (19.116 mg/ml) and ACE inhibition activity with the lowest inhibitory concentration(IC50) value (0.149 mg/ml). These results indicate that extracts obtained from S. clava tunic may potential candidate to reduce diseases caused by various oxidative stresses and hypertension.

The β-carotene biofortified transgenic rice was developed by transforming rice cv. Nakdongbyeo with phytoene synthase (Psy) and carotene desaturase (Crt I) genes isolated from Capsicum and Pantoea. The aim of this study was to perform molecular characterization of rice transformants of T5-T7 generation harboring Psy and Ctr I genes driven by endosperm specific globulin promoter for biosafety evaluation of β-carotene biofortified transgenic rice. The structure and sequence of T-DNA in the transformation vector and the insertion sites, flanking sequences and generational stability of inserted T-DNA in transgenic rice lines were analyzed. The transformation vector consisted of right border, MAR gene, carotenogenic genes unit, herbicide resistance selectable marker unit, MAR gene and left border in sequential order. T-DNA was introduced at the position of 30,363,938-30,363,973 bp of chromosome No. 2 by adaptor-ligation PCR. Stable integration of T-DNA and stable expression of bar gene was confirmed in T5 to T7 generations. It was also confirmed that the backbone DNA of transformation vector containing antibacterial gene was not present in the genome of β-carotene biofortified transgenic rice. HPLC analysis confirmed that carotenoids were consistently detected through T5-T7 generations.

β-카로틴 강화벼 도입유전자의 도입 위치와 안정성을 도입 위치 주변염기서열 분석과 서던 분석한 결과, 벼 염색체 2번 중 30,363,938번과 30,363,973번 사이의 단일 부위에 도입유 전가 도입되었으며, T5-T7 세대 동안 도입된 모든 유전자들 이 안정적으로 유지되고 있으며, 도입유전자의 운반체인 Backbone DNA (pSB11)는 β-카로틴 강화벼에 삽입되지 않 았음을 확인하였다. 선발 마커로 도입된 PAT 단백질의 발현 분석 결과에서도 T5-T7 세대별, 생육시기별로 안정적으로 발 현됨을 검정하였으며, 목적 형질인 β-카로틴 분석 결과에서 도 모품종인 낙동벼에 비해 9.4배 함량이 증가됨을 확인하였 다. 이상의 분석기법을 통해 복수세대에서 β-카로틴 강화벼 의 도입 유전자들이 안정적으로 유지되고 목적 단백질들이 안정적으로 발현되고 있음을 확인하였다.

본 연구는 내충성 독소 발현 유전자(CryIIIA)를 벼에 형질 전환하여 해충에 대한 저항성을 갖도록 국내에서 개발한 해충저항성 GM벼(Btt12R)와 그 모본인 낙동벼의 주요영양성분과 항영양소를 분석하여 각 성분의 함량에 차이가 있는지를 비교하기 위해서 수행하였다. 이를 위해, 낙동벼와 Btt12R 뿐만 아니라 국내 상업화 품종인 영안벼와 화성벼를 수원 GMO 격리포장에서 동일한 조건하에 재배하여 수확한 현미를 사용하였다. 47가지 주요영양성분(8가지 일반성분, 17가지 아미노산, 8가지 지방산, 9가지 미네랄, 5가지 비타민) 중에 16가지 성분의 함량이 모본과 Btt12R 간에 차이를 보였지만, Btt12R의 이 16가지 성분은 함께 재배한 일반벼와 OECD에 명기된 함량 범위 내에 있었다. 2가지 항영양소 중 트립신 저해제는 모든 시료에 0.1 TIU/mg 미만의 극미량으로 존재했으며, Btt12R의 피트산 함량은 낙동벼와 일반벼의 피트산 범위에 포함되었다. 이상의 결과를 종합해보면, 분석한 Btt12R의 모든 주요영양성분 및 항영소의 함량이 모본 및 상업화 품종의 함량 범위에 포함되었으며, 이를 통해 CryIIIA 유전자를 벼 게놈에 삽입하는 것이 현미의 영양학적 품질에 영향을 미치지 않음을 확인하였다.

Keratin wastes are generated in excess of million tons per year worldwide and biodegradation of keratin by microorganisms possessing keratinase activity can be used as an alternative tool to prevent environmental pollution. For practical use of keratinase, its physicochemical properties should be investigated in detail. In this study, we investigated characteristics of keratinase produced by Xanthomonas sp. P5 which is isolated from rhizospheric soil of soybean. The level of keratinase produced by the strain P5 increased with time and reached its maximum (10.6 U/ml) at 3 days. The production of soluble protein had the same tendency as the production of keratinase. Optimal temperature and pH of keratinase were 40℃-45℃ and pH 9, respectively. The enzyme showed broad temperature and pH stabilities. Thermostability profile showed that the enzyme retained 94.6%-100% of the original activity after 1 h treatment at 10℃-40℃. After treatment for 1 h at pH 6-10, 89.2%-100% of the activity was remained. At pH 11, 71.6% of the original activity was retained after 1 h treatment. Although the strain P5 did not degrade human hair, it degraded duck feather and chicken feather. These results indicate that keratinase from Xanthomonas sp. P5 could be not only used to upgrade the nutritional value of feather hydrolysate but also useful in situ biodegradation of feather.

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.