PUBX1和PUBX2基因对拟南芥器官发育的调控
发布时间：2019-06-22 15:19
【摘要】：植物器官的大小的调控是一个基本的发育生物学过程,其生长受发育遗传程序和环境因素的影响,其研究是植物发育生物学领域的重要前沿和研究热点。植物不同物种之间器官大小差异巨大,而特定物种的器官最终大小却是一定的,表明植物器官生长到最终大小受到严密的遗传控制。器官生长受到的时空调节,主要分为细胞分裂和细胞体积扩增两个不同的过程。本项目以模式植物拟南芥为研究材料,通过分子生物学,细胞生物学,生物化学等综合研究手段,对调节拟南芥器官发育生长的重要基因PUBX1及其同源基因PUBX2的功能进行研究,解析高等植物器官大小决定的遗传和分子机制,为植物生长发育的遗传改良提供理论依据。本研究通过对野生型,pubx1,pubx2单突变体以及pubx1 pubx2双突变体的花瓣和叶片的长度、宽度、表面积以及茎的直径的表型进行拍照观察并测量统计,发现单突变体与WT相比无明显变化,而双突变体的表型非常明显,因此PUBX1和PUBX2是有功能冗余的影响器官大小的基因。我们利用Gateway技术构建了PUBX1和PUBX2基因的过表达植株,通过RT—qPCR对其表达量分析可知,PUB基因表达量与植株器官大小呈负相关,表达量越高的植株器官越小。我们同时构建了△PUBX1和△PUBX2基因的过表达植株作为对照,RT—qPCR结果可得表达量明显上调的删除U-box结构域的植株表型与WT相比无显著变化,进一步验证了U-box结构域对于PUBX1和PUBX2基因的酶活是必需的。为了进一步了解PUBX1和PUBX2基因的作用机制,我们构建了表达PUB基因启动子及全长蛋白与GUS报告基因的融合蛋白的转基因植株,筛选阳性植株进行GUS组织染色来研究PUB基因的表达模式,发现其在成熟的花瓣中表达量远远大于幼嫩的花瓣中。同时我们测量统计并绘制了花瓣的生长曲线,得出PUBX1和PUBX2基因是通过调控细胞分裂持续时间进而影响植株器官发育的大小。本研究通过对植物器官大小调控机制的探究,为作物的遗传改良提供理论依据,对植物器官大小调控的机制研究将有助于提高作物产量,解决粮食危机有深远的现实意义。
[Abstract]:The regulation of plant organ size is a basic developmental biological process, its growth is affected by developmental genetic procedures and environmental factors, and its research is an important frontier and research focus in the field of plant developmental biology. There are great differences in organ size among different species of plants, but the final size of organs in specific species is certain, indicating that the growth of plant organs to the final size is closely controlled by heredity. The temporal and spatial regulation of organ growth is mainly divided into two different processes: cell division and cell volume amplification. In this project, Arabidopsis thaliana, a model plant, was used as the research material to study the function of the important gene PUBX1 and its homologous gene PUBX2, which regulate the organ development and growth of Arabidopsis thaliana by means of molecular biology, cell biology and biochemistry. The genetic and molecular mechanism of organ size determination in higher plants was analyzed, which provided a theoretical basis for the genetic improvement of plant growth and development. In this study, the length, width, surface area and stem diameter of petals and leaves of wild type, pubx1,pubx2 single mutant and pubx1 pubx2 double mutant were observed and measured. It was found that there was no significant change in the phenotype of single mutant compared with WT, but the phenotype of double mutant was very obvious, so PUBX1 and PUBX2 were genes with functional redundancy affecting organ size. The overexpression of PUBX1 and PUBX2 genes was constructed by Gateway. RT-qPCR analysis showed that the expression of PUB gene was negatively correlated with plant organ size, and the higher the expression was, the smaller the plant organs were. At the same time, we constructed the overexpression plants of PUBX1 and PUBX2 genes as control. The results of RT-qPCR showed that the phenotypes of plants deleted U-box domain with up-regulated expression had no significant change compared with WT, which further verified that U-box domain was necessary for the enzyme activity of PUBX1 and PUBX2 genes. In order to further understand the mechanism of PUBX1 and PUBX2 gene, we constructed transgenic plants expressing PUB gene promoter and fusion protein of full-length protein and GUS reporter gene. Positive plants were screened for GUS tissue staining to study the expression pattern of PUB gene. It was found that the expression of PUB gene in mature petals was much higher than that in young petals. At the same time, we measured and plotted the growth curve of petals, and it was concluded that PUBX1 and PUBX2 genes affected the development of plant organs by regulating the duration of cell division. Through the study of the regulation mechanism of plant organ size, this study provides a theoretical basis for the genetic improvement of crops, and the study of the mechanism of plant organ size regulation will be helpful to improve crop yield and solve the food crisis.
【学位授予单位】：深圳大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：Q943.2
本文编号：2504703
[Abstract]:The regulation of plant organ size is a basic developmental biological process, its growth is affected by developmental genetic procedures and environmental factors, and its research is an important frontier and research focus in the field of plant developmental biology. There are great differences in organ size among different species of plants, but the final size of organs in specific species is certain, indicating that the growth of plant organs to the final size is closely controlled by heredity. The temporal and spatial regulation of organ growth is mainly divided into two different processes: cell division and cell volume amplification. In this project, Arabidopsis thaliana, a model plant, was used as the research material to study the function of the important gene PUBX1 and its homologous gene PUBX2, which regulate the organ development and growth of Arabidopsis thaliana by means of molecular biology, cell biology and biochemistry. The genetic and molecular mechanism of organ size determination in higher plants was analyzed, which provided a theoretical basis for the genetic improvement of plant growth and development. In this study, the length, width, surface area and stem diameter of petals and leaves of wild type, pubx1,pubx2 single mutant and pubx1 pubx2 double mutant were observed and measured. It was found that there was no significant change in the phenotype of single mutant compared with WT, but the phenotype of double mutant was very obvious, so PUBX1 and PUBX2 were genes with functional redundancy affecting organ size. The overexpression of PUBX1 and PUBX2 genes was constructed by Gateway. RT-qPCR analysis showed that the expression of PUB gene was negatively correlated with plant organ size, and the higher the expression was, the smaller the plant organs were. At the same time, we constructed the overexpression plants of PUBX1 and PUBX2 genes as control. The results of RT-qPCR showed that the phenotypes of plants deleted U-box domain with up-regulated expression had no significant change compared with WT, which further verified that U-box domain was necessary for the enzyme activity of PUBX1 and PUBX2 genes. In order to further understand the mechanism of PUBX1 and PUBX2 gene, we constructed transgenic plants expressing PUB gene promoter and fusion protein of full-length protein and GUS reporter gene. Positive plants were screened for GUS tissue staining to study the expression pattern of PUB gene. It was found that the expression of PUB gene in mature petals was much higher than that in young petals. At the same time, we measured and plotted the growth curve of petals, and it was concluded that PUBX1 and PUBX2 genes affected the development of plant organs by regulating the duration of cell division. Through the study of the regulation mechanism of plant organ size, this study provides a theoretical basis for the genetic improvement of crops, and the study of the mechanism of plant organ size regulation will be helpful to improve crop yield and solve the food crisis.
【学位授予单位】：深圳大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：Q943.2
【参考文献】
相关期刊论文 前3条
1 徐化学;熊建华;傅彬英;;应用Gateway技术构建水稻OsDAD1基因的RNA干涉载体[J];分子植物育种;2007年01期
2 陈其军,安瑞,周海梦,陈珈,王学臣;使用与Gateway技术兼容的T载体获得入门克隆[J];生物化学与生物物理进展;2004年10期
3 ;GATEWAY~(TM)── 一种新型克隆技术[J];微生物学通报;2000年05期
，本文编号：2504703
本文链接：https://www.wllwen.com/kejilunwen/jiyingongcheng/2504703.html
最近更新
教材专著
