Unfavorable environmental stresses are major limiting factors that affect plant productivity. Plants perceive and respond adaptively to an abiotic stress condition, and the adaptive process is controlled mainly by phytohormone, consequently, changing in gene expression pattern. Transcriptional regulator ABF3 (Abscisic acid response element Binding Factor 3) mediating the ABA-responsive gene expression plays important roles in drought and temperature tolerance. Here, we report an event of drought tolerant GM gourd in which abf3 was inserted in the genome. For drought stress experiment, T0 plants were self-pollinated and back-crossed to select cultivars for drought stress experiment. The drought tolerant GM gourds were selected for last 3 years and have been crossed to get the several BC and T generation consecutively. In this year, BC3T1, F1 (Bak x GM gourd), BC1F1 and control plants were subjected to drought stress, that is no watering for 12 days and rehydration afterwards. The GM gourds showed high tolerance to drought while the non-transformed plants were totally dried. When the plants were subjected to rehydration, the GM bottle gourds were completely revived and recovered from the drought stress. Tolerance levels to drought of BC3T1, F1 (Bak x GM gourd) and BC1F1 were 92.5%, 50.0% and 65.0%, respectively.

Bt gene derived from the B. thuringiensis has been used for developing GM crops, and corn, cotton and soybean producing B. thuringiensis toxins have been on the market for last 17 years or so creating a huge GM seed industry. One of the notorious pests in brassica crops is diamondback moth (DBM). In order to protect the insect plague of crops from DBM, 4-5 billion dollars have been wasted annually for applying integrated measures in worldwide. Major prevention is use of pesticides that may build the contamination level of chemicals in the ground and this practice threats the environment and ecosystem. An alternative is to develop GM brassica crops and therefore we have developed GM cabbages resistant DBM using bt gene. Lots of T0 cabbages were tested for resistance and independent GM cabbages resistant to DBM were selected. Molecular analysis was conducted to find if GM cabbage holds one copy transgene and intergenic insertion. We found an independent GM cabbage and it contained a singly copy of the transgene without disturbing the insertion site. This one called C95 line with an status of event have been self-crossed for two generation (T2). Also we are working the development of GM cabbage with different vector that contains bar gene as a selection marker. So far 17 T0 cabbages have been obtained by bar selection.

Many viruses infect cucurbits. One of the well-known symptoms is mosaic disease. Those that cause mosaic are cucumber mosaic virus (CMV), squash mosaic virus (SqMV), watermelon mosaic virus (WMV), zucchini yellow mosaic virus (ZYMV) and cucumber green mottle mosaic virus (CGMMV). WMV resistant GM squash was developed many years ago in the United States and it was on the market, but no further information was available by now pertinent to commercial aspect. Usually these viruses are not easily controlled by frequent applications of chemicals that target the insect as carriers of viruses. Therefore, it is necessary to develop commercial varieties possessing resistance against viral diseases. We have developed GM watermelon rootstocks called gongdae, using a coat protein gene of CGMMV as transgene. Those GM watermelon rootstocks showed highly resistant to CGMMV, and have been crossed to get the several BC and T generation. In order to obtain the virus resistant watermelon, watermelon lines were crossed to the selected GM watermelon rootstock. Here, we present the successful watermelon cultivars that show resistance to CGMMV. The resistance must have obtained by transferring the transgene from the GM watermelon rootstock to watermelon line

CMVP1 (cucumber mosaic virus pathotype 1) has been frequently occurring virus causing damage in pepper farms, and it is hard to control the outbreak due to lack of the genetic source resistant to this specific pathotype. Therefore, we have developed transgenic peppers tolerant of CMVP1 using a CP gene of CMVP0 pathogen. In order to fulfill the requirement of the biosafety assessment criteria, we have studied the horizontal gene flow from GM pepper to non-GM pepper by monitoring the transgene movement. If the pepper farms are located closely each other and the pollen moves from GM pepper to non-GM pepper, it would cause unintended fertilization. Therefore, a buffer zone to separate the cultivation regions is required to avoid the contamination of transgene. Previously, several data regarding the movement distance of pepper pollen were reported by judging the phenotypic change. However, no tool as a trace marker was available. The objective of this study was to assess the frequencies of gene flow from GM peppers to non-GM peppers in neighboring farms using the transgene of CP as a trace marker. The GM and non-GM peppers were cultivated in the isolated farm of Nongwoo Bio Co. (NW GM pepper field) and pepper fruits were collected from the NW GM pepper field as well as the neighboring pepper farms. The pepper seeds collected from the farms were planted and the massive PCR analysis was performed to answer the question how far the pollen of GM pepper migrates. The conclusive data based on the consecutive experiments for 6 years is that the gene flow by pollen movement did not occur in peppers that were separated each other over 30 m.

This study investigated the nutrients and anti-nutrients of produced resveratrol GM rice (Iksan-515, Iksan-526, Dongjin) that were cultivated Iksan and Suwon regions. Among the rice samples, Iksan-515 and Iksan-526 are produced resveratrol GM rice. Reveratrol is health-beneficial compound with strong antioxidant and antitumor activities. Red wine is believed to the main source of resveratrol in the human diet. Recent studies have associated resveratrol with the cardio-protective effect observed among people with moderate resveratrol consumption. Moreover, resveratrol has been possess chemoprotective activity. In present study, we determined the substantial equivalence between GM rice and seedling sort. We investigated the nutrients and anti-nutrients of produced resveratrol GM rice and analyzed nutrients including moisture, crude fat, ash, crude protein, fatty acids, amino acids and minerals. The results of this analysis showed equivalence between GM rice and non-GM rice. We determined phenolic compounds including naringenin, vallin and investigated 4 tocopherols (α-, β-, γ-, δ-tocopherol) and 4 tocotrienols (α-, β-, γ-, δ-tocotrienol). Among the all rice cultivars, they showed substancial equivalence between resveratrol GM rices and non-GM rice.

Genetically modified (GM) plant claims to be the solution to global poverty, and potentially solving environmental change and food requirement by increased human population. In this study, we were evaluating agronomic characteristics and chemical properties of two GM drought-tolerant rice (CaMsrB2-8 and CaMsrB2-23) compared with donor cultivars (Ilmi). Statistical analysis agronomic characteristics GM and donor rice showed no significant difference between both of them. Yield and appearance of rice grain, GM rice was a similar to the donor rice. Chemical composition analysis showed that GM drought-tolerant rice has no different with donor rice. This result indicated that GM drought tolerant rice has no big significant difference agronomic character and chemical properties; it can be solve food shortages in spite of drought condition.

고추(Capsicum annuum L.)는 가지과(Solanaceae)로 분류되는 일년초로서 온대에서 열대까지 널리 재배되고 있다. 또한 세계 50억 인구가 식생활에 활용하고 있고 재배면적이 7위인 세계적인 채소로서 식생 외에도 먹거리의 식품첨가제, 색소, 의약품재료, 화장품 등 산업적으로 많이 사용되고 있으며, 그 산업규모가 계속 증가하고 있다. 고추 관련 산업 매출은 300억불로서 상업적 가치가 매우 높은 채소작물이다. 따라서 고추는 채소류 중 동남아 등 해외로 수출가능성이 가장 높은 작물로써 수출전용 품종 개발이 시급하고, 국내 재배용으로는 바이러스 내병성이 강하고 고품질의 고추 품종 개발이 요구된다. 현대 생물공학은 고추의 병해충에 대한 유전적 개량을 통해 이익을 줄 수 있으며, 첫 번째 유전자 형질전환은 Agrobacterium tumefaciens로 neomycin phosphotransferase와 β-glucuronidase 유전자를 삽입한 것으로 (Liu et al., 1990) 그 후로 C. annuum에 대한 기술의 발달이 꾸준히 보고되었다. 최근 고추에서 개량하고자 하는 형질에는 CMV, TEV, TMV에 대한 바이러스 저항성(Cai et al., 2003), 담배나방(Heliothis assulta)에 대한 해충저항성(Kim et al., 2003), 과실성숙도 조절, 저장기간 연장 등으로 요약되고 있다(OECD 고추표준기술서, 2006). 본 연구에서는 국내에 존재하는 고추 품종과 계통을 모두 이병시키는 새로운 바이러스인 CMVP1에 대한 내성이 있는 GM고추 H15 event의 농업적 특성 포장시험을 실시하였다. 농업환경위해성 평가를 위한 농업적 특성 포장시험을 2012년 5월부터 9월까지 H15 event와 모본 P2377에 대하여 수행하였다. 조사내용은 식물체, 과실 및 꽃의 특성을 조사하였다. 식물체의 특성은 6월부터 9월까지 6항목을 조사하였으며, 8월 최성기는 18항목을 조사하였다. 조사결과는 6월부터 8월까지 식물체 특성은 통계적으로 유의한 차이가 없었으며, 9월에는 P2377 고추의 초폭이 H15 고추에 비해 유의하게 컸지만 초장, 주경장, 경경, 엽장 및 엽폭은 통계적으로 유의한 차이가 없었다. 과실의 특성은 미숙과 20항목과 숙과 19항목을 조사하였으며, 착과상태, 과장, 과경 등 조사대상 항목의 특성은 모두 통계적으로 유의한 차이가 없었다. 꽃의 특성은 화색 등 10가지 항목을 조사하였으며, 화색, 웅예수 등 통계적으로 유의한 차이가 없었다. 종자의 특성은 종자색등 4가지 항목을 조사하였으며, 종자색, 종자크기 등 통계적으로 유의한 차이가 없었다. 이와 같은 연구결과는 포장시험을 통한 농업적 특성의 위해성평가 기초자료로 제시할 수 있을 것이며, GMO식품안정성평가에 대한 가이드라인으로 제시 할 수 있을 것으로 기대된다.

In Korea, CMV (cucumber mosaic virus) is the most frequently occurring virus with a single infection rate of 45%. However, a total occurrence of CMV by co-infection, either couple or multiple, with BBWV (broad bean wilt virus), PepMoV (pepper mottle virus) and PMMoV (pepper mild mottle virus) covers over 90% in the field cultivation of pepper. The PepMoV is transmitted by several aphid species, and it has been considered the most frequently detected potyvirus when it co-infects with CMV or PMMoV. Since F1 hybrid that resistant to PepMoV has not been developed, we have developed transgenic peppers using Agrobacterium-mediated transformation with a Hc-Pro gene of the PepMoV. A large number of T1 peppers were tested for resistance to the PepMoV, and T1 peppers tolerant of PepMoV were selected. After consequent self-crossing up to T4 generation, highly tolerant peppers to PepMoV were selected. So far, BC3F1 lines have been selected by back-crossing with 4 elite lines through a breeding program. The horticultural differences of the GM line comparing to inbred lines were investigated and no statistical significance between GM and non-GM lines was found. Based on molecular analysis, One of GM lines, 10-2, contained the transgene in the non-coding region indicating that this line would be a GM event.

잔디는 비식용, 비사료용 작물이지만 스포츠공간용, 조경용, 지피용 등으로 전 세계적으로 가장 보편적으로 활용되는 작물이며, 경제성이 매우 높은 작물이다. 본 연구의 목적은 상업적 재배를 위해 개발된 제초제저항성 웅성불임 GM들잔디의 분자생물학적 특성을 평가하는 것이다. JG21-MS 웅성불임계통은 최초 선발된 해로부터 현재까지 6여 년 동안 제초제저항성 뿐만 아니라 웅성불임성을 잘 유지 하고 있다. BAR 유전자카세트와 T-DNA 삽입점에서 좌,우 1kb 정도의 주변 염기서열의 동등성 및 BAR유전자 발현의 동등성에서 모본과 차이를 보이지 않았다. T-DNA 삽입특성을 조사한 실험에서, T-DNA 오른쪽 경계(RB)의 삽입은 특이한 DNA단편의 첨가 또는 소실 없이 들잔디 DNA와 연결되어 있지만, hygromycin 저항성 유전자(HPH)가 위치한 왼쪽 경계(LB)의 삽입은 삽입과정에서 HPH 유전자의 4/5 정도가 소실되어 유전자의 기능을 상실하였다. 또한 왼쪽 끝 부분과 연결된 들잔디 DNA 쪽도 620bp가 소실되었다. T-DNA는 Ty1-copia retrotransposon-like 유전자 영역에 삽입 된 것으로 확인되었다. Ty1-copia retrotransposon은 식물 종에 조금 차이가 있지만 전체게놈의 10~50% 정도를 차지하는 것으로 알려져 있다. 또한 retrotransposon-like 유전자는 식물고유기능의 유전자가 아니므로 본 이벤트에서 T-DNA는 비유전자영역에 삽입된 것으로 간주할 수 있을 것으로 사료된다. 결국, JG21과 JG21-MS는 HPH 유전자가 소실되어 기능을 상실하였으므로 목표 유전자인 BAR 만 single copy로 존재하는 것으로 확인되었다. 차세대바이오그린21 GM작물실용화사업단에서 지원하는 GM작물 이벤트의 분자생물학적 특성의 최소기준은 단일복제수(single copy), 항생제마커 없이 목표유전자 도입(target gene only), 비유전자 내 삽입을 장려하고 있고, JG21와 JG21-MS 계통의 제초제저항성 들잔디는 이러한 기본조건을 만족시키고 있는 것으로 평가되었다.

Many viruses infect cucurbits. One of the well-known symptoms is mosaic disease. Those that cause mosaic are cucumber mosaic virus (CMV), squash mosaic virus (SqMV), watermelon mosaic virus (WMV), zucchini yellow mosaic virus (ZYMV) and cucumber green mottle mosaic virus (CGMMV). WMV resistant GM squash was developed many years ago in the United States and it was on the market, but no further information was available by now pertinent to commercial aspect. Usually these viruses are not easily controlled by frequent applications of chemicals that target the insect as carriers of viruses. Therefore, it is necessary to develop commercial varieties possessing resistance against viral diseases. We have developed GM watermelon rootstocks called gongdae, using a coat protein gene of CGMMV as transgene. Those GM watermelon rootstocks showed highly resistant to CGMMV, and have been crossed to get the several BC and T generation. In order to obtain the virus resistant watermelon, watermelon lines were crossed to the selected GM watermelon rootstock. Here, we present the successful watermelon cultivars that show resistance to CGMMV. The resistance must have obtained by transferring the transgene from the GM watermelon rootstock to watermelon line.

Bt gene derived from the B. thuringiensis has been used for developing GM crops, and corn, cotton and soybean producing B. thuringiensis toxins have been on the market for last 16 years or so creating a huge GMO industry. One of the notorious pests in brassica crops is diamond backmoth (DBM). In order to protect the insect plague of crops from DBM, 4-5 billion dollars have been wasted annually for applying integrated measures in worldwide. Major prevention is use of pesticides that may build the contamination level of chemicals in the ground and this practice threats the environment and ecosystem. An alternative is to develop GM brassica crops and therefore we have developed GM cabbages resistant DBM using bt gene. Lots of T0 cabbages were tested for resistance and independent GM cabbages resistant to DBM were selected. Molecular analysis was conducted to find GM cabbage to hold one copy transgene and intergenic insertion. We found two independent GM cabbages as an event and those have been self-crossed for two generation. Also we are working the development of GM cabbage with different vector that contains bar gene as a selection marker.

In 2012, the world population exceeded 7 billion and the need to address food security has never been greater. Achieving food security won’t be easy considering the megatrends of growing population, greater affluence, and increasing urbanization. Not only are more people demanding more food, but they want greater variety, including meat, dairy, fruits and vegetables. While demand for food is growing, farmers’ ability to increase productivity is facing unprecedented challenges. Scarcity of water, energy, and land is expected to define food production in the coming decades. Agricultural practices will also need to protect biodiversity through increasing productivity without expanding into natural ecosystems. Further exacerbating the situation is a changing climate that has led to higher temperatures and erratic weather patterns in some areas. Each day farmers face the challenge of growing more from less - increasing yields while protecting the environment by using less water, land, and energy. Global Agricultural Biotechnology companies like Syngenta have addressed these challenges through innovation in research and development by looking at the grower’s challenges holistically, including land, technology, and the community. The presentation will cover general R&D activities in an agricultural biotechnology company, which may differ from those in academic research institutes. Product safety and environmental considerations are integral to industry’s R&D work. To make earlier and better-informed decisions on which active ingredients or traits to move forward, normally companies begin safety trials early in the development process, facilitating timely engagement with regulators and other key stakeholders. Also to complement in-house expertise and bring in novel technologies which may or may not be used in agribusiness, companies are actively seeking value-adding partnerships and collaborations to bring exciting new offers to the grower. Development of a GM crop through all those activities mentioned above is quite a costly and lengthy process. My presentation will describe a typical process required for developing a GM crop in an agricultural biotechnology company from early discovery to commercialization to the market, which may give you a different perspective from academic point of view.

Daily shedding pattern and longevity of pollen are important consideration for the evaluation of gene flow of transgenic plants. During the day, the pollen shedding pattern of zoysiagrass was determined in the lawn by using a device to collect airborne pollen on a glass slide, resulting that the pollen grains were released predominantly between 7:00 and 9:00. The result was also supported by in vitro pollen germination test, which was performed with pollens collected from 1:00 through 24:00 at 1h interval. Influence of temperature and humidity on pollen longevity was determined by germinating pollen at 25°C after incubating them for 10, 30, 60, and 180 min under different temperatures and humidity with pollen of zoysiagrass that opened freshly at about 9:00. The result showed that pollen longevity of zoysiagrass was sensitive to change of temperature and humidity and longest under the temperture and humidity of 15-20°C and 80-99%, respectively. Under natural conditions with the same method as upper controlled conditions, was determined pollen longevity. Under sunny atmospheric conditions, pollen longevity decreased to 20% in 60 min, with a complete extinction in 120 min. Under cloudy atmospheric conditions, pollen remained viable up to 450 min, with about 20% longevity after 360 min. No significant difference was found between GM and non-GM plants in their pollen longevity.

Bt gene derived from the B. thuringiensis has been used for developing GM crops and those crops are already on the market. The aim of this study is to construct a genetic transformation with peppers using CryIAc1 gene and consequently to develop GM peppers resistant to the oriental tobacco budworm. We have developed transgenic peppers using Agrobacterium-mediated transformation, and obtained 5 T0 peppers. T0 peppers were self-crossed and T1 pepper fruits were exposed to larvae to test the survival rate. The survival rate of larvae that were fed with GM fruits decreased dramatically while with non-GM fruits the rate was not changed much. In order to establish the selection and the culturing of bug through a year, an incubating system for tobacco budworm in chamber was manufactured and a selection system under the media that are mixed with GM green pepper was obtained. Using those system, T3 peppers tolerant to tobacco budworm were selected.

A Transgenic Kimch cabbage has been developed harboring T-DNAs expressing delta-endotoxin insecticidal protein, herbicide (basta) resistant protein, and antisense transcript of AsMADS2 gene. Three transgenic lines, #24, #45, and #51, originated from the same T0 plant were analyzed in terms of molecular characterization, phenotype, and agronomic traits. Flanking sequence analysis confirmed that T-DNA, with 7132 bp intact structure, was inserted onto the pseudochromosome A10 of B. rapa and all the genes in T-DNA were functionally active. Three of GM cabbage showed 69.2～75.3% of plant height and 81.8～89.7% of diameter to those of the isogenic variety ‘Nowon’, respectively. Curving upward leaf lamina attitude was observed on GM cabbage, while straight or slight concave on non-GM cabbage. In addition, an average range of 86～91.5% of head height and 87.4～94.8% of head diameter were observed on GM cabbage to those of the isogenic variety ‘Nowon’, respectively Moreover, curled inwards or slight overlap of head-forming leaf overlap at terminal region was observed on GM cabbage, but curled outwards or erect on non-GM cabbage. AsMADS2, a transcription factor reported to be involved in early flowering, was stably expressed to RNA in the GM cabbage, but it was not shown the significant influences to flowering time.

The significance of genetic stability and bio-safety environment has been recently recognized by many GM plants. This study was to evaluate the GM stability of transgenic rice and to identify the environment variance. The GM rice of vitamin A　-enriched rice and four check cultivars were analyzed the data on agronomic characters and principal component for 2009-2011 in large-GM crop field. Cultivation environment was conducted in the large-GM field and greenhouse to determine grain characters. In this experiment, there was no significant difference in agronomic characters between GM rice of vitamin A-enriched rice and a donor plant, Nagdong. Related to grain characters, grain appearance and physicochemical characteristics were similar to GM rice of vitamin A-enriched rice and a donor plant, Nagdong. However, grain appearance in GM rice of vitamin A-enriched rice showed to white core and white belly when GM rice of vitamin A-enriched rice was planted in greenhouse. The type and distribution of dominant weed species also were not different from GM rice of vitamin A-enriched rice and a donor plant, Nagdong. Additionally that of gene flow was not detected in dominant weed species by PCR analysis.

Plant breeding has at this moment two gene pools available: 1. plant breeders’ gene pool consisting of all crossable germplasm. All the genes from this source are available in classical plant breeding. Functional genes isolated from this natural source are called “cisgenes”; 2. a new gene pool consists of transgenes, which are chimeric and mostly partly or totally consisting of genes from non-crossable species, including bacteria, viruses, etc. When GM breeding started in the eighties of last century’ only transgenes belonging to the new gene pool were available. The complicated biosafety regulations, needed for this new gene pool, have been based on these so called transgenic traits but because of the transformation process accidentally they are also including cisgenes. In this way cisgenesis belongs to the expensive class of GM breeding which is only practiced by multinationals in the so called large crops such as maize, soybean and cotton. Reconstructed logic delivers several arguments against the classification of cisgenesis into the GMO class: 1. Cisgenes are already existing in nature and belong to the breeders gene pool, 2. It does not fit the definition of a GMO, 3. It is in practice classical breeding replacing existing introgression and translocation breeding with the advantage of absence of linkage drag, 4. The EFSA recently showed that cisgenesis is as safe as classical plant breeding, 5. Another EU committee on new techniques concluded in majority that a sequence of at least 20 bp is needed to come to a new combination which has to be classified as GMO. So, cisgenic plants, mostly without insertion of borders, are not considered to be a GMO. A simple rule should be developed, including criterions, for defining true cisgenic plants. In this presentation, the creation of cisgenic, more durable, resistance of potato against potato late blight will be discussed. This is based on working simultaneously on the genetics of both potato and Phytophthora infestance and on stacking of broad spectrum R-genes. Isolation and use of over 20 R-genes and more than seven Avr-genes will be described and the use of them to come to functional stacking of at least three R-genes. Another important issue is, because of absence of cisgenic selection markers, the setup of a marker free transformation system. Cisgenesis is the most effective way to improve in one step worldwide frequently used free potato varieties, which are highly susceptible to late blight. If needed additional broad spectrum R-genes can be added later by re-transformation.