Agrobacterium을 이용한 탐미 유채의 형질전환 체계 개발
Development of Agrobacterium-mediated transformation system in Brassica napus 'Tammi'
- 주제(키워드) 도움말 Rapeseed , Plant regeneration , Plant growth regulator , Acetosyringone , AgNO3
- 발행기관 강릉원주대학교 일반대학원
- 지도교수 도움말 박남일
- 발행년도 2020
- 학위수여년월 2020. 8
- 학위명 석사
- 학과 및 전공 도움말 일반대학원 원예학과
- 실제URI http://www.dcollection.net/handler/kangnung/000000010715
- UCI I804:42001-000000010715
- 본문언어 한국어
초록/요약 도움말
Rapeseed, a cruciferous plant such as Chinese cabbage and mustard, is an oil crop containing about 38 to 45% oil in the seeds. About 15-20% of rapeseed oil is soluble nitrogen and about 20% is edible oil containing protein, which is consumed the most after soybean oil. Rapeseed is used as a raw material for biodiesel with high commercial value because it contains about 60-70% of oleic acid in addition to edible oil. In plant tissue culture technology, the efficiency of re-differentiation differs according to the species, and even in the same species, various results can be obtained depending on variety, the type of culture tissue, and the type and concentration of plant growth regulators added to the culture medium. In this experiment, I aim to establish an efficient plant re-differentiation method and a transformed tissue culture system for Tammi rapeseed. In addition, the characteristics of transformants by the insertion of the Adote1 gene were observed. In preliminary experiments, the most efficient concentration of plant growth regulators to induce re-differentiation from explants was the combination of 0.5 mg/L naphthalene acetic acid (NAA) and 8 mg/L kinetin. The frequency of re-differentiation was about 56.5%, and 12 subjects were selected by the medium with 50 mg/L kanamycin. The target gene was confirmed to be inserted in the selected individuals through PCR, and the final Tammi rapeseed transforments were selected. Seed size of the transformants was smaller compared to wild type, and the role of Adote1 gene in rapeseed was confirmed through transformation.
more목차 도움말
목 차
초록 ⅴ
I. 서언 1
II. 연구사 3
1. 유채 3
2. 세계 유채 재배 및 생산 추이 5
3. 국내 유채 재배 및 생산 추이 8
4. 국내 유채 육종 품종 12
5. 식물 형질전환 연구 현황 15
6. 식물 형질전환 기술 16
7. Agrobacterium을 이용한 유채의 형질전환 연구 17
8. 본 연구에 사용된 유전자 19
III. 재료 및 방법 20
1. 식물재료 준비 20
2. 형질전환용 Adote1 유전자 및 Agrobacterium 배양 21
3. 외식편 접종 및 공배양 22
4. 세척 및 선발 22
5. 뿌리유도 23
6. 식물체 순화 23
7. PCR을 통한 형질전환체 확인 24
IV. 결과 25
1. 형질전환 배지에서 AgNO3의 역할 25
2. 재분화를 위한 적정 식물생장조절물질 26
3. 형질전환체 선발을 위한 항생제의 적정 농도 29
4. 형질전환체 생산 32
5. PCR을 통한 유전자 도입 확인 34
6. 형질전환 식물체 종자의 형태적 특성 35
V. 결론 37
VI. 참고문헌 38
