发根转化体系在苦瓜三萜皂苷合成基因研究中的应用

	发根转化体系在苦瓜三萜皂苷合成基因研究中的应用; Application of Hairy Root Transformation System in the Study of Triterpenoid Saponin Biosynthesis Gene of Bitter gourd (Momordica charantia)
	李艳
导师	杨雪飞
摘要	The hairy root system has been widely applied in the researches of gene and secondary metabolites exploration. Characterized by rapid growth, no requirement of exogenous plant hormones, stable genetic traits, and strong ability to synthesize secondary metabolites, it is considered as an ideal culture system for producing secondary metabolites. Triterpenoid saponin is an important category of secondary metabolites found in plants, with a variaty of significant biological activities and a wide range of pharmacological effects. Nevertheless, low provision of triterpenoid saponin limited its wider application, due to its low abundance in plants for direct extraction, and its complex chemical structure for chemical synthesis. New synthetic apprpaches are therefore in needed. Bitter gourd (Momordica charantia) containing cucurbitane triterpenoids, is a global well-known plant with dual uses as food as well as medicine. Recently, the completion of the draft sequencing of the bitter gourd genome has provided the opportunitiy for biosynthesis and gene-function analysis of its triterpenoids. In this thesis, we aimed at establishing and optimizing a bitter gourd hairy root transformation system, with further attempts to explore the relationship between the OSCs gene and the synthesis of triterpenoid saponin. The major contents and results are as following: （1）Establishment of hairy root transformation system of bitter gourd In order to establish an effective hariry root transformation system, we explored and optimized the conditions on the processes of seed sterilization, selection of infected explants, and callus cultures. The results showed that: 1) it was best to sterilize the seeds with a 4% sodium hypochlorite solution for 3-5 minutes, under which the seed germination rate was the highest and the contamination rate was the lowest; 2) in the selection of explants, cotyledons cotyledon with petiole was better than cotyledons and cotyledons petiole, with shorter rooting time and more hariy roots produced; 3) the explants of bitter gourd can be infected directly, with no requirement of callus culture. The optimized procedure of establishing transformation system of bitter gourd hairy roots includes: seeds to sterilize with 4% sodium hypochlorite for 3-5min, wash with sterile water and inoculate the MS medium to obtain sterile seedlings, cut out the cotyledon with petiole as explants for direct infection, inoculate on MS medium and grow to hairy root growth. This process takes 15-17 days. （2）Identification and analysis of bitter gourd OSCs genes In order to test whether the constructed bitter gourd hairy root system can be applied to the study of the relationship between genes and metabolites, we investigated preliminary the role of bitter gourd OSC gene on the regulation of bitter gourd triterpenoid saponins. Based on the published OSCs genes and homology analysis, the OSCs of bitter gourd were screened and re-identifed according to the conserved domain
	2020-05
文献类型	学位论文
条目标识符	http://ir.kib.ac.cn/handle/151853/74149
专题	昆明植物所硕博研究生毕业学位论文
推荐引用方式 GB/T 7714	李艳. 发根转化体系在苦瓜三萜皂苷合成基因研究中的应用, Application of Hairy Root Transformation System in the Study of Triterpenoid Saponin Biosynthesis Gene of Bitter gourd (Momordica charantia)[D],2020.

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