黄瓜赤霉素受体基因CsGID1a的功能验证与表达谱分析

发布时间：2018-01-29 04:51

  本文关键词： 黄瓜 赤霉素受体 果实心室 瓠果　出处：《中国农业大学》2017年博士论文　论文类型：学位论文

【摘要】：植物激素赤霉素(GA)是高等植物体内的一类四环二萜羧酸化合物,在植物各个生长发育阶段起着至关重要的作用。前人已在赤霉素促进种子发芽,调控植物营养生长和生殖生长等方面开展了较多研究,但有关其调控植物果实发育方面的分子机制尚不明确。本文以黄瓜为试材,通过时空表达、功能验证和表达谱分析,研究了 GA受体蛋白CsGID1a对黄瓜果实心室形成的调控机制,旨在丰富黄瓜果实发育的理论基础,取得的主要结果如下:(1)黄瓜CsGID1a和CsGIID1b属于高等植物赤霉素受体基因GID1家族的同源基因,均含有受体基因所特有的HGG和GXSXG两个保守域,在黄瓜的茎尖和根尖处高度表达,外源GA3处理可导致反馈调节,使CsGID1a和CsGID1b的表达量下调,表明CsGID1a和CsGID1b均具备赤霉素受体基因的典型特征。(2)进化树分析表明CsGID1a处于双子叶植物GID1主效基因ac族,时空表达分析显示CsGID1a在根尖、茎尖、雌花芽及果实中的表达显著高于CsGID1b,干扰CsGID1b表达并不会影响植株生长,表明CsGID1a可能是主效基因。(3)通过原位杂交和GUS染色在花和果实中对CsGID1a的表达进行更详细分析表明,CsGID1a在黄瓜雄蕊、雌蕊及果实的心室壁上特异表达。通过黄瓜遗传转化获得的CsGID1a干扰转基因植株,与对照相比,干扰植株明显矮化,果实心室发育不完全。表达谱分析表明在干扰转基因植株中,生长素合成与运输基因和细胞周期相关基因的表达量都显著低于对照组;同时激素测定与细胞切片观察的结果表明,CsGID1a干扰转基因植株果实中生长素的分布和果实细胞分化受到影响。(4)在拟南芥中的功能验证发现,异源表达CsGID1a可部分恢复双突变体gid1agid1c的植株高度以及植物育性表型,同时也对果实心室的发育造成了影响,部分拟南芥转基因株系的果实还形成了类似黄瓜果实的三心室。
[Abstract]:The plant hormone gibberellin (GA) is a kind of tetracyclic diterpenoid carboxylic acid compounds in higher plants, which plays an important role in the growth and development of plants. More studies have been carried out on the regulation of plant growth and reproductive growth, but the molecular mechanism of regulating plant fruit development is not clear. In this paper, cucumber was used as the material and expressed in time and space. The regulation mechanism of GA receptor protein CsGID1a on the ventricular formation of cucumber fruit was studied in order to enrich the theoretical basis of cucumber fruit development. The main results are as follows: (1) Cucumber CsGID1a and CsGIID1b belong to the GID1 family of gibberellin receptor genes in higher plants. There were two conserved domains, HGG and GXSXG, which were specific to the receptor gene. They were highly expressed at the stem tip and root tip of cucumber. Exogenous GA3 treatment could lead to feedback regulation. The expression of CsGID1a and CsGID1b was down-regulated. The results showed that both CsGID1a and CsGID1b had the typical character of gibberellin receptor gene. The phylogenetic tree analysis showed that CsGID1a was in the ac group of GID1 major genes in dicotyledonous plants. Spatio-temporal expression analysis showed that the expression of CsGID1a in root tip, stem tip, female flower bud and fruit was significantly higher than that in CsGID1b. Interfering with CsGID1b expression did not affect plant growth. The expression of CsGID1a in flower and fruit was analyzed in more detail by in situ hybridization and GUS staining. CsGID1a was specifically expressed in the ventricular wall of stamen, pistil and fruit of cucumber. CsGID1a interference transgenic plants obtained from cucumber genetic transformation were significantly dwarfed compared with the control. The expression profile analysis showed that the expression of auxin synthesis and transporter genes and cell cycle related genes were significantly lower in the transgenic plants than in the control group. At the same time, the results of hormone determination and cell section observation showed that CsGID1a interfered with the distribution of auxin and the differentiation of fruit cells in transgenic plants, and the function of CsGID1a was verified in Arabidopsis thaliana. The heterologous expression of CsGID1a could partially restore the plant height and fertility phenotype of the double mutant gid1agid1c, and also affect the development of fruit ventricle. The fruits of some Arabidopsis transgenic lines also formed three ventricles similar to cucumber fruits.
【学位授予单位】：中国农业大学
【学位级别】：博士
【学位授予年份】：2017
【分类号】：S642.2

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