환경 스트레스 내성 유전자 ABF3 도입 국화 형질전환체 개발 및 특성 분석
Characterization of transgenic chrysanthemum plants horboring stress tolerance gene ABF3
- 발행기관 상명대학교 대학원
- 지도교수 형남인
- 발행년도 2017
- 학위수여년월 2017. 2
- 학위명 석사
- 학과 및 전공 일반대학원 식물식품공학과식물식품공학전공
- 실제URI http://www.dcollection.net/handler/smuclib/000002333029
- 본문언어 영어
초록/요약
Through Agrobacterium-mediated transformation of leaf explants of chrysanthemum (Chrysanthemum morifolium cv. Jinba), transgenic plants expressing the ABF3 gene, a drought tolerance ABF3 gene isolated from Arabidopsis, under the control of the constitutive CaMV 35S promoter (CA plants) or the stress-inducible IbSWPA2 promoter (SA plants) were generated and their characteristics were analysed. Under the selection with phosphinothricin (PPT), 43 and 19 independent transgenic lines of CA and SA plants were selected and the insertion of transgenes into the transgenic plants was identified using PCR analysis Based on PCR analysis, the transformation efficiencies were calculated as 3.6% for CA plants, and 1.9% for SA plants. The copy number of the inserted T-DNA was determined for the 12 independent transgenic lines of CA and SA plants by qPCR analysis of the bar gene. This result indicates the presence of single or two copies of T-DNA insertion in the genomes of the ABF3 transgenic lines. Immuno-strip assay verified stable integration and expression of the bar gene on transgenic plants at the protein level. Among the CN and SN transgenic lines generated, the transgenic lines 1, 3 of CA plants and 3, 8 of the SA plants were selected for the evaluation of tolerance to drought stress. When the plants were grown in greenhouse under normal conditions, compared with the WT plants (25.5 cm), CA plants (20.8 cm) exhibited reduced plant height but SA plants (23.8 cm) displayed a little differences. The WT and ABF3 transgenic lines of CA and SA plants were subjected to 13 days of water-limiting treatment, followed by 5 days of re-watering. After drought treatments for 13 days, the group of CA lines height increased to 13.1% and the group of SA lines increased to 13.4% in plant, and the plant height of WT was increased to 2.1% in comparison to before drought treatment. The photosynthetic performance of WT plants are lower than those of the CA and SA plants during drought treatment period. Because of the dehydration of leaves, WT plants finally died. After re-watering, the photosynthetic performance were recovered a little in the CA and SA plants. In comparing the photosynthetic performance of the WT and ABF3 plants, the drought treatment caused a larger harmful affection in the WT plants than in ABF3 plants. Before and after drought stress treatment, ABF3 gene expression had increased in SA lines, while that of CA lines showed little increase. When the water loss rates from detached leaves of the WT and ABF3 transgenic plants were investigated, the transgenic plants showed slower water loss rates than WT. The proportions of stomatal closure in the ABF3 transgenic plants were much higher than WT plant in the absence or presence of ABA treated conditions. These results showed that ABF3 gene was functional in chrysanthemum and its overexpression appeared to enhance drought in transgenic plants. Furhtermore, ABF3 can be used to improve stress tolerance to drought in many other agriculturally important crops.
more목차
Abstract i
Contents iv
List of Tables v
List of Figures vi
I. Introduction 1
II. Literature Review 3
III. Materials and Methods 14
IV. Results and Discussion 27
V. Abstract in Korean 49
VI. Literature Cited 51