This experiment was conducted at Suwon, Korea from 2013 to 2015. The objective of this study was to establish the optimum seeding rate, and to clarify the nitrogen fertilizer level for rye seed production in central and north area of Korea. We used Korean rye cultivar ‘Gogu’ for this test. We employed a split-plot design with three replications. The main plots were designed by three seeding levels (3, 5 and 7 kg 10a-1), but other sub-plots were randomly seeded. The plots were treated with three different nitrogen fertilizer levels (3, 6 and 9 kg 10a-1). The percentage of productive tiller, number of grain per spike, fertility rate, 1 liter weight, and 1000-grain weight decreased as seeding rate increased from 3 kg 10a-1 to 7 kg 10a-1, whereas the number of spike per ㎡ increased. Therefore the grain yields of rye had less of an effect by increasing seeding rate. There was an increase in number of spike per ㎡, number of grain per spike, and fertility rate as nitrogen fertilizer level increased from 3 kg 10a-1 to 9 kg 10a-1, but grain yields significantly not affected by the interaction of seeding rate × nitrogen fertilizer levels. However, the best seeding rate and nitrogen fertilizer level for rye seed production were 5 kg and 5∼6 kg 10a-1, respectively, considering seed and fertilizer reduction and the prevention of pollution by excess fertilization.

‘Saeyoung’, a winter triticale (X Triticosecale Wittmack) for forage, was developed at the Department of Rice and Winter Cereal Crop, NICS, RDA in 2012. The cultivar ‘Saeyoung’ has narrow and long leaves of light green color, middle size and thin culm, and a medium grain of brown color. The heading date and yellow ripe stage of ‘Saeyoung’ was May 3 and May 27, which were similar to check cultivar ‘Shinyoung’, respectively. ‘Saeyoung’ showed a little stronger in cold tolerance and a little weaker in resistance to lodging than the check, and wet injury, powdery mildew, and leaf rust were similar to those of the check cultivar. The forage fresh and dry matter yields of ‘Saeyoung’ at milk-ripe stages were 47.2 and 15.6 MT ha-1, respectively, which was 9% and 4% higher than those of the check. The crude protein content of ‘Saeyoung’ was 0.4% lower than 6.8% of the check, while was higher than the check cultivar ‘Shinyoung’ in neutral detergent fiber, acid detergent fiber. Total digestible nutrients of ‘Saeyoung’ was also 3% lower than 62.8% of the check cultivar. It showed grain yield of 4.1 MT ha-1, which was 11% higher than that of the check. ‘Saeyoung’ is recommended for fall sowing forage crops in areas in which average daily minimum mean temperatures in January are higher than -10 ℃.

A new naked oat cultivar, ‘Jungmo2005’(Avena nuda L.), was developed for food and forage use by National Institute of Crop Science, RDA in 2010. It was derived from a cross between F1[‘Early80’/‘Gwiri33’] and F1[‘Early80’/‘Gwiri23’]. ‘Early80’, a covered oats, has early heading and high yielding, while ‘Gwiri23’ of covered type and ‘Gwiri33’ of naked type has early heading with large-size grain. ‘Jungmo2005’ has the characteristics of narrow and long leaves of pale green color, middle diameter culm of yellow color and medium grain of whitish yellow color. Cultivar ‘Jungmo2005’ had 2 days earlier heading date (May 9) than the check cultivar ‘Seonyang’ (May 11) in field condition. The ‘Jungmo2005’ showed better winter hardiness than that of the check cultivar, and similar to the check cultivar in respect to lodging resistance. The ‘Jungmo2005’ had 105 cm of culm length, 21.5 cm of spike length, 658 spikes per m2, 82 grains per spike, 23.3 g of 1,000-grain weight, and 622 g of test weight. The grain yield of ‘Jungmo2005’ was averaged 3.38 MT ha-1, which was 6% higher than that of the check. Average forage fresh and dry matter yield of ‘Jungmo2005’ harvested at milk-ripe stage were 44.8 and 12.6 tone ha-1, respectively, compared with 47.5 and 12.5 tone ha-1 of the check. The protein content of the ‘Jungmo2005’ was similar to the check (7.6% and 7.5%, respectively), while ADF (28.6%) and NDF (51.5%) were lower than the check (31.4% and 57.0%, respectively). TDN content and RFV were higher than those of the check (66.3%, 120.3 and 64.1, 105.2, respectively). ‘Jungmo2005’ is recommended for fall sowing cropping only in the south area where daily minimum mean temperatures are averaged higher than -4 ℃ in January, and it should not be cultivated in mountain areas, where frost damages is likely to occur. The areas would do better only to sow in spring season.

‘Shinseong’, a winter forage triticale cultivar (X Triticosecale Wittmack), was developed at the Department of Rice and Winter Cereal Crop, NICS, RDA in 2015. The cultivar ‘Shinseong’ was selected from the cross RONDO/2*ERIZO_11//KISSA_4/3/ ASNO/ARDI_3//ERIZO_7 by CIMMYT (Mexico) in 1998. Subsequent generations were handled in pedigree selection programs at Mexico from 1999 to 2004, and a line ‘CTSS98Y00019S-0MXI-B-3-3-5’ was selected for earliness and good agronomic characteristics. After preliminary and advance yield testing in Korea for 3 years, the line was designated ‘Iksan47’. The line was subsequently evaluated for earliness and forage yield in seven locations, Jeju, Iksan, Cheongwon, Yesan, Gangjin, Daegu, and Jinju from 2013 to 2015 and was finally named ‘Shinseong’. Cultivar ‘Shinseong’ has the characteristics of light green leaves, yellow culm and spike, and a medium grain of brown color. The heading date of cultivar ‘Shinseong’ was April 24 which was 3 days earlier than that of check cultivar ‘Shinyoung’. The tolerance or resistance to lodging, wet injury, powdery mildew, and leaf rust of ‘Shinseong’ were similar to those of the check cultivar. The average forage dry matter yield of cultivar ‘Shinseong’ at milk-ripe stages was 15 MT ha－1, which was 3% lower than that (15.5 MT ha－1) of the check cultivar ‘Shinyoung’. The silage quality of ‘Shinseong’ (6.7%) was higher than that of the check cultivar ‘Shinyoung’ (5.9%) in crude protein content, while was similar to the check cultivar ‘Shinyoung’ in acid detergent fiber (34.6%), neutral detergent fiber (58.6%), and total digestible nutrients (61.6%). It showed grain yield of 7.2 MT ha－1 which was 25% higher than that of the check cultivar ‘Shinyoung’ (5.8 MT ha－1). This cultivar is recommended for fall sowing forage crops in areas in which average daily minimum mean temperatures in January are higher than －10℃.

국립식량과학원에서 국내 환경에 적합한 조사료용 호밀 품종을 개발하기 위하여 1995년에 ‘조춘호밀’ 등 10개 품종을 방임수분으로 혼합 교배하여 그 중에서 형질이 우수한 ‘SR95POP-S1-140-9-1-3-7-5-3’ 계통을 선발하였다. 선발 계통에 대해 생산력을 검정한 결과 특성이 우수하여 ‘호밀50호’로 계통명을 부여하고 5개 지역에서 3개년간 지역적응성을 검정하였으며, 조사료 수량뿐만 아니라 종실 수량이 높은 것으로 나타나 2013년 직무육성신품종 선정위원회에서 신품종으로 선정됨과 함께 ‘씨드그린’으로 명명되었다. ‘씨드그린’은 초형이 직립이고, 잎색은 농록색이며, 길이는 길고, 폭은 넓은 편이다. 줄기는 황백색으로 중간 정도의 굵기이며, 종실은 갈색이고, 중간 크기이다. ‘씨드그린’의 출수기는 4월 22일로 표준품종인 ‘곡우’ 보다 3일, 성숙기는 6월 16일로 2일 빠르다. 초장은 141 cm로 ‘곡우’보다 3 cm 길다. 한해는 평균 2로 표준품종인 ‘곡우’의 1과 대등한 수준이며, 도복은 ‘곡우’ 보다 약간 강하였으며, 습해와 병해는 발생하지 않았다. ‘씨드그린’의 생초수량은 ha당 39.8톤(곡우 40.8톤), 건물수량은 평균 8.3톤(곡우 8.4톤)으로 ‘곡우’ 보다 약간 적었으나, 종실수량은 4톤으로 ‘곡우’보다 16% 증수하였다. ‘씨드그린’의 조단백질 함량은 10.5%로서 ‘곡우’ 보다 0.8% 높았으며, ADF와 NDF 함량(%)은 각각 38.8%와 66.4%로 ‘곡우’ 보다 낮았으나, TDN 함량(%)은 58.3%로 ‘곡우’ 보다 1.3% 높았으며, TDN 수량 또한 4.81 톤/ha로 ‘곡우’ 보다 약간 높았다.

해충저항성 Bacillus thuringiensis (Bt) 벼의 비표적곤충인 벼물바구미(Lissorhoptrus oryzophilus)에 대한 성충 전용살충제 Clothianidin 액상수화제의 살충제 감수성 시험을 실시한 결과, 72시간-LC50은 0.245 ml/L(95% 신뢰한계는 0.195~0.307 ml/L)이었으며, Bt벼의 모본으로 대조로 사용한 낙동벼의 72시간-LC50은 0.257 ml/L(95% 신뢰한계는 0.199~0.331 ml/L)이었다. 72시간-LC50은 낙동벼에서 다소 높았지만, 해충저항성 Bt벼 72시간-LC50이 낙동벼의 95% 신뢰한계 내에 포함되어, 두 품종의 LC50값에 유의성이 없는 것으로 판단된다.

해충저항성 Bt벼와 낙동벼의 미꾸리(Misgurnus anguillicaudatus) 와 잉어(Cyprinus carpio)에 대한 급성독성시험을 실시한 결과 48시간 및 96시간-LC50은 1,000mg/L 이상으로 나타났다. 48시간 및 96시간 무영향농도(NOEC)는 1,000mg/L이었다. 급성독성 시험기간 중 해충저항성 Bt벼와 낙동벼간의 pH, DO, 수온, 체중 및 전장에 대한 유의적인 결과는 나타나지 않았다.

‘Jungmo2501’ (Avena sativa L.), a winter oat for forage use, was developed by the breeding team at the National Institute of Crop Science, RDA in 2010. The following is the characteristics of ‘Jungmo2501’ that is characterized as light green leaf, yellow brown culm and whitish yellow grain. The heading date of ‘Jungmo2501’ was about 3 days earlier than that of check cultivar ‘Samhan’(May 7 and May 10, respectively). Its plant height was 11 cm longer than 103 cm of the check, and the leaf blade ratio of aerial parts was 26 % higher than the check (11.8% and 9.4%, respectively). The cold tolerance, resistance to lodging and wet injury of ‘Jungmo2501’ were similar to those of the check. The average forage dry matter yield of ‘Jungmo2501’ harvested at milk-ripe stage was 5% higher than the check (15.5 ton ha -1 and 14.7 ton ha -1 , respectively). ‘Jungmo2501’ was higher than the check in terms of protein content (6.6% and 5.9%, respectively), neutral detergent fiber (58.5% and 57.6%, respectively), and acid detergent fiber (34.5% and 32.1%, respectively), while total digestible nutrients was lower than the check (61.6% and 63.6%, respectively), and TDN yield was 0.37 ton ha -1 more than that of the check (9.71 ton ha -1 and 9.34 ton ha -1 , respectively). The silage grade of ‘Jungmo2501’ estimated by Flig score showed level Ⅱ, meaning good quality. Fall sowing cropping of ‘Jun

A variety of genetically modified (GM) crops have been developed in Korea. In these crops, the resveratrol-enriched transgenic rice plant has moved ahead to generate the dossier for regulatory review process required for commercialization of GM crop. The resveratrol-enriched transgenic rice plant could be released to farmers for cultivation after national regulators have determined that it is safe for the environment and human health. Here we developed a PCR-based DNA marker based on flanking sequences of transgene for the discrimination of zygosity in resveratrol-enriched transgenic rice plant. This DNA marker will be useful for identifying of resveratrol-enriched transgenic rice plant, and can also be use to estimate transgene movement occurred by pollen transfer or seed distribution.

The β-carotene biofortified transgenic soybean was developed recently through Agrobacterium -mediated transformation using the recombinant PAC (Phytoene synthase-2A-Carotene desaturase) gene in Korean soybean (Glycine max L. cv. Kwangan). GM crops prior to use as food or release into the environment required risk assessments to environment and human health in Korea. Generally, transgenic plants containing a copy of T-DNA were used for stable expression of desirable trait gene in risk assessments. Also, information about integration site of T-DNA can be used to test the hypothesis that the inserted DNA does not trigger production of unintended transgenic proteins, or disrupt plant genes, which may cause the transgenic crop to be harmful. As these reasons, we selected four transgenic soybean lines expressing carotenoid biosynthesis genes with a copy of T-DNA by using Southern blot analysis, and analyzed the integration sites of their T-DNA by using flanking sequence analysis. The results showed that, T-DNA of three transgenic soybean lines (7-1-1-1, 9-1-2, 10-10-1) was inserted within intergenic region of the soybean chromosome, while T-DNA of a transgenic soybean line (10-19-1) located exon region of chromosome 13. This data of integration site and flanking sequences is useful for the biosafety assessment and for the identification of the β-carotene biofortified transgenic soybean.

The β-carotene biofortified transgenic soybean was developed recently through Agrobacterium-mediated transformation using the recombinant PAC (Phytoene synthase-2A-Carotene desaturase) gene in Korean soybean (Glycine max L. cv. Kwangan). GM crops prior to use as food or release into the environment required risk assessments to environment and human health in Korea. Generally, transgenic plants containing a copy of T-DNA were used for stable expression of desirable trait gene in risk assessments. Also, information about integration site of T-DNA can be used to test the hypothesis that the inserted DNA does not trigger production of unintended transgenic proteins, or disrupt plant genes, which may cause the transgenic crop to be harmful. As these reasons, we selected four transgenic soybean lines expressing carotenoid biosynthesis genes with a copy of T-DNA by using Southern blot analysis, and analyzed the integration sites of their T-DNA by using flanking sequence analysis. The results showed that, T-DNA of three transgenic soybean lines (7-1-1-1, 9-1-2, 10-10-1) was inserted within intergenic region of the soybean chromosome, while T-DNA of a transgenic soybean line (10-19-1) located exon region of chromosome 13. This data of integration site and flanking sequences is useful for the biosafety assessment and for the identification of the β-carotene biofortified transgenic soybean.

기존의 유전자변형식물은 외래의 도입유전자를 갖고 있으며 이들로부터 기인한 단백질 또는 합성물질에 의한 의도적/비의도적 영향에 대한 안전성 논란이 사회적 이슈가 되어 왔다. 최근의 기술적 진보에 의하여 이른바 식물육종의 신기술이 발달하게 되었고 이들 기술로 만든 신규식물에 대한 안전성평가에 GMO 관련 규제의 적용 여부 문제가 대두되게 되었다. 이들 NPBTs 기술로 만든 신규식물의 특징은 SDN이나 ODM과 같이 염색체상의 정확한 위치에 짧은 염기서열의 indel(s)이나 단일염기 돌연변이를 도입하여 자연적인 돌연변이와의 구별이 거의 불가능하거나, cisgenesis와 같이 성적교잡이 가능한 종 유래의 유전자를 구조변형 없이 도입하여 근연종과 동질적인 식물을 만들거나, heterozygous 형질전환체 후대세대의 null-segregant 선발이나 epigenetic를 이용하여 도입유전자가 존재하지 않지만 목적 형질을 갖는 식물체를 만드는 장점이 있다. 또한 grafting이나 Agro-infiltration 등의 방법으로 안전성평가를 회피하거나 경감할 수 있는 가능성을 높이게 되었다. OECD를 비롯한 주요 GMO 개발국의 관련 학회에서는 SDN, ODM 및 cisgenesis 또는 intragenesis 기술로 만든 식물에 대하여 non-GM 식물과 동일한 위해성평가 규정을 적용하거나 상황에 따라 완화된 규정을 적용할 수 있다고 판단하고 있다. 현 시점에서 이들 NPBTs 기술을 이용하여 개발된 식물이 상업화된 예는 없으나 많은 국가에서 상업화를 목적으로 개발 중이며 일부에서는 안전성평가를 완료한 단계이다. 이러한 현실에서 NPBTs 기술의 개념 정립, 신규식물의 안전성평가의 방향 설정 및 현실성 있는 작물개발 방안을 마련하여야 한다. 이를 통하여 GMO에 대한 안전성 논란과 사회적 거부감을 우회하는 동시에 답보 상태에 있 는 분자육종 분야의 발전을 도모할 계기를 마련하여야 한다.

GM벼 OsCK는 벼 유래의 OsCK1 유전자를 벼에 형질전환 하여 벼흰잎마름병 및 벼도열병에 대한 저항성을 높게 한벼로 농촌진흥청에서 개발하였다. 형질전환 벡터의 구성은 양쪽 border (LB, RB) 상간에 2개의 MAR 염기서열이 서로 마주보는 형태로 위치하고 있으며, 제초제 저항성 유전자 PAT는 CaMV 35S promoter에 의하여 발현이 유도되고, 목표 유전자인 choline kinase (OsCK)는 actin promoter에 의하여 발현이 조절되며 left border 기준으로 역방향으로 배치되었다. 도입유전자 확인을 위하여 adaptor ligation PCR을 수행하였는데, MAR 영역에 위치하는 제한효소로 GM벼 genomic DNA를 절단한 후 adaptor를 붙였다. 염기서열 분석을 위하여 T-DNA의 양 말단에서 primer를 제작한 후 sequence 분석을 하였다. 분석한 결과, T-DNA의 right border 인근의 MAR sequence가 벼 genome의 10번 염색체 129971번 염기와 연결되어 있음을 확인하였다. Left 영역의 삽입위치는 이후 실시한 Illumina NGS 시스템을 이용하여 확인할 수 있었으며, GM 벼에는 2개의 T-DNA가 도입되었음을 알 수 있고, 첫 번째 T-DNA는 벼 10번 염색체 BAC클론 OSJNBa0014J14의 128947번째 염기와 129970째 염기에 위치하고 벼 genome 염기 1024 bp가 결실됨을 확인하였다. 이 과정에서 첫 번째 T-DNA left border와 첫 번째 MAR sequence 일부(370 bp)가 결실되었고 right border와 두 번째 MAR 영역 199 bp가 결실되었음도 확인하였다. 두 번째 T-DNA는 right border가 결실된 형태로 첫번째 T-DNA의 35S promoter 중간에 삽입되었음을 확인하였다.

Two carotenoid biosynthetic genes, phytoene synthase (Psy) and carotene desaturase (CrtI) linked via synthetic 2A sequence under control of CaMV 35S promoter (two T0 plants 5 and 6) or β- conglycinin promoter (three T0 plants 7, 13 and 16) were transformed into soybean variety Kwangan. After agronomic and phenotypic selection at early generations, T5 progeny of PAC soybean were analyzed by Southern blot to confirm T-DNA copy numbers. A total of 27 homologous lines derived from one of three T0 plants (line 7 under the control of β- conglycinin promoter) with one copy T-DNA insertion, were separated and planted into greenhouse. Flanking sequence analysis was carried out on one of homologous line 6-2-3 and results indicated the T-DNA was intergenic inserted into chromosome 14 from 10,873,131 to 10,872,998 base of soybean chromosome. T-DNA insertion structure, flanking sequence and inserted gene expressions need to be analyzed in the further study.

Resveratrol rice Iksan526 was developed by overexpession of T-DNA (RB::P-Ubi::RS::T-NOS::P-35S::PAT::T-35S::LB) in rice variety Dongjin. To confirm one locus insertion of T-DNAs, Mendelian genetic analysis was carried out on selection marker bar gene and objective RS gene separately by using a F2 population derived from a cross of Dongjin/Iksan526 (T6). A total of 450 four-leaf-old plants from F2 population were treated by 0.3% basta, and a phenotypic separation ratio of 3:1 (321 survival: 129 dead, p>0.90) complied with Mendelian inheritance indicating one locus insertion of bar gene. Genotypic separation was analyzed by using PCR with specific primers for 300 plants, which were selected from 321 survival plants after phenotypic separation. Results revealed a ratio 1:2 of homologous to heterozygous (92:208, p>0.90), which further confirmed one locus insertion of RS gene. In addition, comparison on agronomic traits and resveratrol contents between transgenic rice and the donor variety were launched to evaluate the phenotypic performance over multi-generations (years).

Tocopherols (α-, β-, γ- and δ-tocopherols) represent a group of lipophilic antioxidants which are synthesized only by photosynthetic organisms. It is widely believed that protection of pigments and proteins of photosynthetic system and polyunsaturated fatty acids from oxidative damage caused by reactive oxygen species (ROS) is the main function of tocopherols. In the present study, NtTC, which encodes a tobacco tocopherol cyclase ortholog, was cloned and characterized. Compared with control plants, NtTC transgenic rice showed higher tolerance to drought stress, and total tocopherol content increased by 52 % in leaf. Additionally, total antioxitant activity of NtTC transgenic lines was increased significantly by 19%. These results demonstrate that over-expressing NtTC could improve the tolerance to abiotic stress in rice, and tocopherols play a crucial role in the protection of oxidative stress.

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 rice water weevil (RWW), Lissorhoptrus oryzophilus are major pests of aquatic rice plant in Korea as well as throughout the country. Larvae of RWW sucking the nourishment on roots, causes a stunted root system and reduces grain yields. To prevent these damages, we constructed various plant expression vectors, which were harbored by insecticidal genes, cryBP1 and cryIIIa, and fused with the actin promoter and/or the modified RCg2 root-preferential promoters for expressing the insect-toxic genes in leaves and roots. A cryBP1 was cloned from Bacillus popilliae, producing crystal toxin against Japanese beetle, and CryIIIa was modified from the δ-endotoxin gene of Bacillus thuringiensis ssp. tenebrionis, encoding the coleoptera-specific toxin. The vectors containing the insecticidal genes were transferred into Oryza sativa japonica cultivar, Nakdong, by Agrobacterium -mediated transformation method. Several independent transgenic lines were selected by Southern blotting and Western blotting, confirming that cryBP1 and cryIIIa genes were stably integrated into the plant genomes and were expressed in transgenic plants. Upon insect bioassay using RWW, the mortality of insect larvae on cryBP1 and cryIIIa transgenic rice lines recorded up to 41% and 34%, respectively. These results suggested that the transgenic lines can be used to develop Coleoptera-resistant cultivars and could be valuable for later application in crop breeding for insect resistance.