브로꼴리 엽록체 ATP Synthase Subunit 1 유전자의 5' Exon 부위 cDNA 클론 및 이의 발현 양상

서미정(Mi Chung Suh),이한길(Han Kuil Yi),홍주봉(Choo Bong Hong) 한국식물학회 1995 Journal of Plant Biology Vol.38 No.2

We isolated a cDNA clone, BLSC1, encoding 5` exon of ATP synthase CF_0 subunit I from broccoli. BLSC1 is 285 nucleotides long which consists of a 5` noncoding region of 34 nucleotides, a 5` exon of 145 nucleotides and an intron of 106 nucleotides. The 5` exon codes for 48 amino acids which reveals mostly hydrophobic. The amino acid sequence deduced from BLSC1 shares 83%, 83% and 91% identities with the genes coding for atpF from wheat, rice and spinach, respectively. Genomic Southern blot analysis for BLSC1 showed a typically strong signal for a gene located in the chloroplast genome. Northern blot analysis identified three major classes of transcripts showing strong positive signals in the leaves, but only trace amounts of the transcripts were identified in the other organs like stems, flower buds and roots.

유채의 $F_1$ 종자 생산시 종자친과 화분친의 재식비 및 등숙시기별 종자의 지방산 분석

구양규,양선영,정용수,김현성,서미정,안성주,Ku, Yang-Gyu,Yang, Sun-Young,Jung, Yong-Su,Kim, Hyun-Sung,Suh, Mi-Chung,Ahn, Sung-Ju 한국작물학회 2010 한국작물학회지 Vol.55 No.3

이 실험의 목적은 유채 $F_1$ 종자 생산시 종자친과 화분친의 재식비 및 수확시기별 종자의 지방산 함량에 영향을 미치는지 알아보고자 실험을 수행하였다. 안정적인 종자 생산을 위해, 두 가지의 실험을 노지에서 수행하였다. 실험 1은 $F_1$ 종자 생산을 위한 종자친과 화분친의 재식 비별(4:2, 10:2, 10:1)로 정식하였고 유채 개화 후 40, 45, 50, 55, 60에 지방산 함량을 조사하였다. 실험 2는 $F_2$ 종자의 개화 후 등숙시기별로 종자를 수확하여 지방산을 분석하였다. 실험의 결과, 종자친과 화분친 재식비에 따른 등숙시기별 유채 종자의 지방산 조성 및 함량은 차이를 보여 주었으나 지방산의 오염 정도 척도인 erucic acid는 재식비에 따라 영향을 미치지 않았다. 종자친과 화분친의 비에서 얻어진 $F_1$과 $F_2$ 종자 비에서 palmitic acid, stearic acid과 linoleic acid은 등숙 시기에 따라 감소하였다. 그러나 oleic acid는 두 실험 모두에서 개화 후 55일까지 상대적으로 증가하였지만, 개화후 60일째 영향을 미치지 않았다. 지방산 생합성 관련 유전자인 SAD, FAD1 및 FAD2의 발현양상도 등숙 시기별 지방산 조성 및 함량과 일치하였다. 이런 결과는 종자친과 화분친의 비에 따른 재식비과 $F_2$종자의 비에 따른 지방산 조성이 큰 문제가 되지 않음을 보여주므로 $F_1$ 종자의 안정적인 생산을 위해 종자친의 파종비는 늘리고 화분친비를 줄이면 유채 종자 생산량이 높아질 것으로 사료가 된다. The objective of this experiment was determined to investigate the effect of the planting ratio of Male Sterility (MS) to Restorer Line (RL) and harvesting time on fatty acid compositions under $F_1$ seed production of Brassica napus L. For rapeseed seed production, two experiments were conducted in the open fields. One experiment studied planting ratios of MS to RL (4:2, 10:2, or 10:1) were planted and investigated fatty acid composition at 40, 45, 50, 55, and 60 days after flowering, the other $F_2$ seeds were analyzed on fatty acid compositions of harvested seeds at five sequential stages. The results showed that fatty acid compositions of developing seeds were influenced by MS:RL planting ratios and $F_2$ hybrid treatments and contaminated level of fatty acid compositions, erucic acid, were unaffected by planting ratio of MS to RL. Fatty acid compositions such as palmitic acid (C16:0), stearic acid (C18:0) and linoleic acid (C18:2) contents decreased during seed maturation period in $1^{st}$ and $2^{nd}$ experiments. In contrast, oleic acid (C18:1) content relatively increased up to 55days after flowering. At day 60 after flowering, oleic acid content was unaffected by MS:RL planting ratios and $F_2$ seeds treatments. Aspects of related gene expression of fatty acid synthesis such as SAD, FAD1 and FAD2 were followed exactly to changes of fatty acid composition during seed maturation. These results suggest that MS ratio may be enlarged and RL may be reduced, indicating this ratio will be useful for rape seed production.

한국 고유의 품종을 이용한 제초제 저항성 유채 개발

김효진,이혜진,고영삼,노경희,이영화,장영석,서미정,Kim, Hyo-Jin,Lee, Hye-Jin,Go, Young-Sam,Roh, Kyung-Hee,Lee, Young-Hwa,Jang, Young-Seok,Suh, Mi-Chung 한국식물생명공학회 2010 식물생명공학회지 Vol.37 No.3

화석에너지의 고갈과 지구 온난화 현상으로 인해 재생 가능한 식물자원으로부터 바이오에너지를 얻고자 하는 관심이 높아지고 있다. 이에 바이오디젤의 원료로 사용 되기 적합한 형질전환이 된 유채를 개발하기 위한 첫 단계로 한국 고유 유채 품종을 이용한 형질전환 체계를 구축하였다. 내한, 영산, 탐미, 한라 유채의 종자를 분양 받아 지방산 분석을 실시한 결과, 종자의 약 32-40% 식물성 오일이 포함되어 있었고, 그 중 올레인산의 함량은 60mole% 이상 존재하는 것으로 확인되었다. 그 중 오일 함량 및 올레인산 함량이 높고, 형질전환 효율이 비교적 높은 한라 유채품종이 그리고 $\beta$-glucuronidase (GUS)와 phosphinothricin acetyltransferase (PAT) 유전자가 포함된 pCAM-BIA3301 벡터가 도입된 Agrobacterium tumefaciens GV3101균주가 형질전환에 사용되었다. 형질전환이 된 유채 식물체는 제초제에 대한 내성, PCR을 이용한 PAT 유전자의 도입 여부 및 GUS 활성 분석을 통하여 선별하였다. 그 결과 한라 유채의 경우, 10. 4% 형질전환 효율을 보였고, 제초제 저항성이 다음 세대 ($T_1$ 식물체)로 안정되게 유전됨을 확인하였다. 이러한 연구는 바이오디젤 원료로 사용될 다양한 유채 품종에 교배를 통해 제초제 저항성 유전자를 쉽게 도입할 수 있는 가능성을 제시하였다. An interest in the production of seed-oil based fuel and raw materials, which comes from renewable plant sources, has been intrigued by the phenomenon of global warming and shortage of fossil fuels. Rapeseed (Brassica napus) is the most important oilseed crop, which produces seeds with 40% oil. It is desirable to develop genetically modified rapeseed producing oils, which can be easily converted to biodiesel. As an initial step for development of genetically modified rapeseed for the production of biofuels or bio-based materials, Korean rapeseed cultivars, Naehan, Youngsan, Tammi and Halla, were analyzed. Four Korean rapeseed cultivars produce 32 to 40% oil of seed dry weight, which is rich in oleic acid (more than 60 mole%). The cotyledonary petioles of rapeseed cultivar, Halla, were transformed using Agrobacterium tumefaciens strain GV3101, carrying the uidA gene encoding $\beta$-glucuronidase (GUS) as a reporter gene and the phosphinothricin acetyltransferase (PAT) gene as a selectable marker. The stable integration of PAT gene in the genome of transgenic rapeseeds was confirmed by PCR analysis. Expression of uidA gene in various rapeseed organs was determined by fluorometric assay and histochemical staining. Transformation efficiency of a Korean rapeseed Halla cultivar was 10.4%. Genetic inheritance of transgenes was confirmed in $T_2$ generation.

카멜리나 (Camelina sativa Crtz.) 발아 적온 및 발아초기 뿌리생육 특성

박준성(Joon Sung Park),최영인(Young In Choi),김용휘(Augustine Yonghwi Kim),이상협(Sang Hyub Lee),김경남(Kyung-Nam Kim),서미정(Mi Chung Suh),김기준(Gi-Jun Kim),이긍주(Geung-Joo Lee) 충남대학교 농업과학연구소 2013 농업과학연구 Vol.40 No.3

A genus Camelina has been attracted as a promising oil crop, especially available in drought and marginal conditions. Due to more demands on arable land for bioenergy crops, price of agricultural products has been a challengeable issue. In that respect, development of Camelina crop with higher germination rate and germination energy can be a strategy to secure seedling establishment, nutrient uptake and long vegetative period. In order to be easily available in the field and laboratory conditions, Camelina seed needs to be optimized for its germination temperature. Germination temperature regime was in a range of 8 to 32℃ initially, and consecutively narrowed down to 8 to 20℃. Based on the temperature range, Camelina germinated greater than 96% at 8-16℃ in two weeks after sowing, but germination rate started to decrease at the higher than 24℃ and was significantly low at higher than 32℃. In terms of rapid time to reach the maximum germination rate and greater germination energy, temperature ranged from 12 to 16℃ was found to be desirable for Camelina germination. Although germinationa rate was greater at 16℃, lower temperature close to 12℃ would be favored for the field conditions where greater root growth leading to healthier seedlings and better nutrient or water availability is considerably demanded.

Physiological Response, Fatty Acid Composition and Yield Component of Brassica napus L. under Short-term Waterlogging

Yang-Gyu Ku(구양규),Won Park(박원),Jin-Ki Bang(방진기),Young-Seok Jang(장영석),Yong-Beom Kim(김용범),Hyun-Jong Bae(배현종),Mi-Chung Suh(서미정),Sung-Ju Ahn(안성주) (사)한국생물환경조절학회 2009 시설원예‧식물공장 Vol.18 No.2

단기간 침수처리에 의한 유채의 영양쟁장과 생식생장기의 생리적 반응, 지방산 조성과 수확량에 관하여 조사하였다. 영양생장 단계에서 10일과 생식생장 단계에서 4일 침수한 결과를 영양생장과 생식생장기 모두 광합성, 기공전도도와 증산량은 침수처리 기간과 회복시기에 현저하게 감소하였다. 침수처리가 끝난 처리구에서 광합성과 기공전도도는 2~3일 내에 대조구와 비슷한 경향을 보였다. 침수처리된 작물의 지방산 조성은 대조구에 비해 통계적 유의성의 차이를 보이지 않았지만 수확량과 관련된 요인들에서 현저하게 감소함으로 생식생장 단계에서의 단기간 침수처리가 수확량 요인에 큰 영향을 줌으로서 종지 수확량에 영향을 미칠 것으로 사료된다. The effects of short-term waterlogging on physiological responses, fatty acid compositions and yield components of rapeseed at both the vegetative growth and the reproductive growth stages were assessed in this study. Waterlogged plants were treated for a period of 10 days at the vegetative growth stage and 4 days at the reproductive growth stage. The results show that photosynthesis and stomatal conductance at both the vegetative growth and the reproductive growth stage were significantly reduced during the waterlogging period and after the recovery period. When the plants were removed from water, photosynthesis and stomatal conductance progressively restored similar values to those of control plants within 2~3 days. Fatty acid compositions were unaffected by waterlogging treatment. However, yield components (pod number and pod length) of the waterlogged treated plants at the reproductive growth stage were significantly reduced. These results suggest that short-term waterlogging may thus influence oilseed yield component.

영산 유채를 이용한 형질전환체 생산

노경희 ( Kyung Hee Roh ),곽보경 ( Bo Kyoung Kwak ),김현욱 ( Hyun Uk Kim ),이경렬 ( Kyeong Ryeol Lee ),김순희 ( Sun Hee Kim ),서미정 ( Mi Chung Suh ),김효진 ( Hyo Jin Kim ),김종범 ( Jong Bum Kim ) 한국응용생명화학회 2011 Journal of Applied Biological Chemistry (J. Appl. Vol.54 No.1

To improve genetic transformation of Brassica napus winter cultivar ``Youngsan``, factors influencing shoot regeneration and transformation from cotyledonary petioles were investigated. Shoot induction was enhanced in the combination of 0.5 mg/L NAA and 2~4 mg/L kinetin. Silver nitrate was essential for successful shoot regeneration, ranging from 5 to 9 mg/L. The addition of GA3 promoted plant regeneration. Among the tested Agrobacterium strains, co-cultivation times, and antibiotic selection regimes, choice of appropriate Agrobacterium strain was the most critical factor for efficient transformation of B. napus cv. ``Youngsan``. The EHA105 succinamopine strain was the most efficient and the maximum transformation efficiency was 26.8%. Transgenic shoots were selected on 10 mg/L phosphinothricin (PPT) containing media. The transgenic plants expressing bar and gus genes were resistant for commercial herbicide Basta and stained with X-Gluc. Southern blot hybridization indicated that the presence of one to three gus gene copies per genome and inheritance of the gus gene into the T1 generation.

Transformation of Tobacco(Nicotiana tabacum) for Hygromycin-Resistance

KIM, Young Soon,SOH, Woong Young,SUH, Mi Chung,HONG, Choo Bong 全北大學校 基礎科學硏究所 1994 基礎科學 Vol.16 No.-

Agrobacterium을 사용하여 hygromycin phosphotransferase 유전자를 식물체에 도입하기 위하여 vector를 제조합하였다. 식물발현 vector인 pBKS-1에서 neomycin phosphotransferase유전자를 제거한 다음, CaMV의 35S프로모터와 nopaline synthase의 3' poly A signal 사이에 HPH유전자를 삽입하였다. 이 hygromycin저항성 유전자는 담배 세포에서 선발 marker로서 기능하였다. 비형질 전환체는 저농도의 hygromycin 5 mg/L에서 생장이 저해된 반면, 형질전환체는 hygromycin 50 mg/L를 함유하는 배지상에서 선발되어 완전한 식물체로 분화되었다. Northern hybridization 방법으로 형질전환체에 도입된 HPH유전자의 발현을 확인하였다. we constructed a vector for the integration of a hygromycin phosphotransferase gene into plant genome by Agrobacterium mediated transformation. A bacterial coding sequence for the hygromycin phosphotransferase was inserted into a plant expression vector, pBKS-1, between CaMV 35S promoter and the nopaline synthase 3' polyadenylation signal after deleting the neomycin phosphotransferase gene from pBKS-1. The bacterial resistance gene to hygromycin B was expressed as a dominant selection marker in tobacco cells. Transformants could be selected in 50 mg/L hygromycin while the untransformed cells were nomally sensitive to 5mg/L hygromycin. Northern hybridization confirmed the presence and expression of the hygromycin phosphotransferase gene in the transformed plants.