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형질전환된 감자의 후대 식물체에서 CaMV 35S Promoter - GUS 유전자 발현
성순기(Soon Kee Sung),최상봉(Sang Bong Choi),전종성(Jong Seong Jeon),박민철(Min Chul Park),이광웅(Kwang Woong Lee) 한국식물학회 1994 Journal of Plant Biology Vol.37 No.1
Two potato (Solanum tuberosum L.) cultivars were transformed by Agrobacterium tumefaciens harboring cauliflower mosaic virus (CaMV) 35S promoter and β-glucuronidase (GUS) gene. Expression patterns of the CaMV 35S promoter according to tissue types and deveplopmental stages, and genetic stability of GUS gene were investigated in the clonal progenies of transgenic potatoes. Kanamycin-resistant shoot emerged from tuber disc after 4 weeks of culture, and root was induced 6 weeks after culture on the selection medium. Shooting frequency of cvs. Superior and Dejima were 43% and 27%, respectively. Mature transformants and their clonal progenies showed no phenotypical abnormality. GUS activity was expressed primarily at parenchymatous cells of phloem tissue around the vascular cambium in the stem and root, and higher activity was found at the apical meristem of shoot, root and adventious shoot bud. GUS activity was higher at tubers of young explants than at stored tubers. These facts indicate that expression level of the CaMV 35S promoter differred according to tissue types and developmental stages of the organs. The GUS gene was stably inherited to each clonal progeny and normally expressed.
반수체 담배의 엽육 원형질체로부터 MNNG 처리에 의한 Glyphosate 저항성 클론의 선별
성순기(Soon Kee Sung),최상봉(Sang Bong Choi),이광웅(Kwang Woong Lee) 한국식물학회 1993 Journal of Plant Biology Vol.36 No.1
Selection of glyphosate-resistant clones from MNNG-treated mesophyll protoplasts of haploid tobacco and their differentiation were studied. The protoplasts were treated with 0.1 to 100 ㎛/mL N-methyl-N`-nitro-N-nitrosoguanidine (MNNG) for 30 min when they expanded to oval shapes. After the treatment, the protoplasts in 4-16 cell stages were transferred to the selective medium containing 1 mM glyphosate for the selection of the glyphosate-resistant colonies. The efficiency of the cell division of the protoplasts in the selective medium decreased as the MNNG concentrations increased. Optimal MNNG concentration for induction of the glyphosate-resistant clones was 10 ㎛/mL and mutation frequency was 2.66 × 10 exp(-6). The stability of the glyphosate-resistance of the clones was examined by prolonged subculture in the medium with 1 mM glyphosate, and the resistant clones were survived more than 10 months. Among them one clone has been proliferating and greening and the others were proliferating without greening or greening with slower proliferating.
입자사출기법에 (粒子射出技法) 의한 벼 세포로의 외래 유전자 도입과 그 발현
전종성(Jong Seong Jeon),정호성(Hou Sung Jung),성순기(Soon Kee Sung),이종섭(Jong Seob Lee),최양도(Yang Do Choi),김한집(Han Jip Kim),이광웅(Kwang Woong Lee) 한국식물학회 1994 Journal of Plant Biology Vol.37 No.1
For establishing a transformation system of rice, an efficient introduction of foreign genes into embryogenic cell suspension by particle bombardment was conducted. The particle inflow gun based on the acceleration of DNA-coated tungsten particles using pressurized helium was constructed for delivery of DNA into rice cells. Several bombardment parameters were optimized using the transient expression of GUS gene. The conditions that gave the highests GUS gene expression of about 1000 blue spots per a g fresh weight of bombarded cells include treatment of the cells with 0.5 M osmotic pressure, and use of the 410 kPa helium, 100 mm target distance, 13 mm syringe filter holder and 5 μL DNA/tungsten mixtures. It was also confirmed that rice actin promoter-intron constract gave the highest expression of all promoter-sequences studied. Eight weeks after the bombardment, stably transformed calluses were obtained on the selection medium containing 100 mg/L G418 and showed the strong activity in in situ GUS assay.
옥잠화(Hosta plantaginea Aschers.)의 개화습성, 수분양식 및 결실특성
조규원(Cho Kyoo Won),태경환(Kyoung-Hwan Tae),성순기(Soon-Kee Sung) 한국자원식물학회 2005 한국자원식물학회지 Vol.18 No.2
본 연구는 원예 및 조경용 식물로 최근 들어 관심을 받고 있는 옥잠화에 대한 개화습성, 수분 양식 및 결실특성에 대한 기초정보를 얻고자 수행되었다. 옥잠화는 주로 야간에 백색꽃이 개화하는 식물로 개화초기와 개화성기에는 대부분 풍매에 의해 수정되었고, 개화후기로 갈수록 착화 간격이 줄어들고 옥잠화 특유의 향기가 감소하여 풍매에 의한 착협율과 충매에 의한 수정율이 증가하는 것으로 조사되었다. 착협율과 결실율은 방임구 각각에서 33.3%와 24.3%로 나타났으며, 이에 비해 인공교배는 각각 65.0%와 40.9%로 나타나 인공교배가 착협율과 결실율에서 높게 나타났다. 개화기간에 있어서 하루 중 수정율은 개화성기인 19:00~20:00시에서 높았으며, 이때의 최적온도는 25~28℃ 이었다. This study was carried out to obtain the basic informations on the flowering habit, pollination patterns and seed setting in Hosta plantaginea Aschers. Flowers came out in the night with white color. Almost fertilization seemed to be out by wind in early and full flowering. But bearing pods by insects and fertilization rate by wind were increased by interval of flowering and particular scent slowly decreased in after flowering. Bearing pods and seeds setting rate were 65.0%, 40.9% per peduncle after artificial pollination, and also 33.3%, 24.3% in natural pollination, respectively. The flowering times was all around seven to eight o clock at night and optimum temperature for pollination was 25~28℃.
서미숙(Mi-Suk Seo),조철오(Chuloh Cho),정남희(Namhee Jeong),성순기(Soon-Kee Sung),최만수(Man-Soo Choi),진민아(Mina Jin),김둘이(Dool-Yi Kim) 한국자원식물학회 2021 한국자원식물학회지 Vol.34 No.4
유전자 가위 기술 등 생명공학 기술을 콩에 적용하여 새로운품종을 개발하기 위해서는 효율적인 조직배양 기술이 필수적이다. 식물의 유전형은 조직배양 효율에 의존하는 형질전환 기술의 성공 여부를 결정짓는 중요한 요소로 알려져 있다. 본 연구에서는 우리나라 콩 핵심 집단 내 21개 자원들을 선발하여, 외래품종 2종과 함께 조직배양 효율을 조사하였다. 그 결과, 근연 관계가 높은 Kwangan, Anpyeong, Seonam은 발아율과 재분화효율이 높았으며, Daepung, Daewon 품종은 발아율과 재분화율 모두 낮게 관찰되었다. 또한 3종의 외래 품종에서는 표준 유전체 해독에 사용된 Williams82와 Jack, Maverick 모두 높은조직배양 효율을 보였다. 조직배양 효율이 높은 자원들을 대상으로 Agrobacterium법에 의한 형질전환을 수행하여 PCR 및bar-strip 분석한 결과 Kwangan, Pungwon, Seonam, 그리고Maverick 품종에서 제초제 저항성 유전자의 삽입을 확인할 수있었다. 이들 결과를 바탕으로 농업적 가치가 높은 다양한 콩 품종들의 형질전환을 통한 새로운 품종 개발이 가능할 것이다. Efficient in vitro regeneration system is essential for the successful crop breeding of soybean (Glycine max (L.) Merr.) using the new biotechnology. The genotype of donor plants strongly influences the establishment of tissue culture system. Therefore, the screening of genotypes with excellent tissue culture ability is very important for soybean genetic improvement. In this study, we report the tissue culture efficiency of 21 soybean cultivars belong to Korean soybean core-collection and two foreign cultivars (Jack and Maverick). The Kwangan, Anpyeong and Seonam are share close genetic relationship in 21 cultivars and these three cultivars were observed the high frequency of germination and regeneration. Furthermore, the high tissue culture abilities were also observed in the Williams 82 used in reference genome sequencing and the two foreign cultivars. The transformation of pBAtc:tRNA with bar gene was performed by Agrobacterium tumefaciens in the cultivars with high tissue culture ability. Transformation of the bar gene was identified by PCR analysis in Kwangan, Pungwon, Seonam, and Maverick. Our results provide useful information for the breeding of various soybean cultivars by plant biotechnology such as, genome editing.