水母雪莲表皮毛发育相关基因SmTTG1的遗传转化研究

发布时间：2018-05-24 13:42

  本文选题：水母雪莲 + 表皮毛　； 参考：《青海大学》2017年硕士论文

【摘要】：水母雪莲(Saussurea medusa Maxim),又称水母雪兔子,属菊科风毛菊属,是一种生长于青藏高原地区的珍稀、濒危药用植物。同时,它又是一种典型的高山植物。植株密被表皮毛是水母雪莲最具有代表性的形态特征,它是水母雪莲对极端环境的一种适应结构。研究水母雪莲表皮毛的形成和发育过程,对揭示高山植物适应环境的特殊机制将具有重要的理论意义。为此,本研究以水母雪莲作为实验材料,以表皮毛发育相关基因SmTTG1为研究对象,构建了SmTTG1基因的植物表达载体,通过根癌农杆菌为介导,转入普通烟草中。主要研究结果如下:(1)根据本实验室已克隆得到的水母雪莲SmTTG1基因序列(Genbank登陆号:KX447632),在其CDS两侧分别设计上下游引物,在上游引物5′端加入酶切位点SalⅠ,在下游引物5′端加入酶切位点Bam HⅠ,克隆了水母雪莲SmTTG1基因的CDS序列(含酶切位点)。(2)通过酶切法,将目的基因SmTTG1插入质粒pRI 101-AN DNA的多克隆位点,构建了植物表达载体pRI 101-AN-SmTTG1,并通过冻融法将其导入农杆菌C58C1中。(3)在根癌农杆菌介导下,通过叶盘法将目的基因SmTTG1转入普通烟草中。通过PCR和RT-PCR检测,获得8株转基因烟草植株。形态学比较表明,转基因烟草植株与对照植株无明显差异,说明SmTTG1基因过量表达不一定对烟草的形态产生影响。
[Abstract]:Saussurea medusa Maxima, also called jellyfish snow rabbit, is a rare and endangered medicinal plant in the Qinghai-Tibet Plateau. At the same time, it is a typical alpine plant. The dense coat of the plant is the most representative morphological feature of the jellyfish Saussurea, which is an adaptive structure of the jellyfish to the extreme environment. It is of great theoretical significance to study the formation and development of the epidermal hair of Saussurea jellyfish in order to reveal the special mechanism of adaptation of alpine plants to the environment. Therefore, the plant expression vector of SmTTG1 gene was constructed by using Saussurea jellyfish as experimental material and epidermal development related gene SmTTG1 as the research object. The vector was transferred into common tobacco by Agrobacterium tumefaciens mediated by Agrobacterium tumefaciens. The main results are as follows: (1) according to the SmTTG1 gene sequence of Saussurea jellyfish cloned in our laboratory, the upstream and downstream primers were designed on the two sides of CDS, and the restriction site Sal 鈪，

本文编号：1929261