拟南芥pny突变体分析及沙柳PNY基因克隆与组织特异性表达分析
发布时间：2019-01-05 16:55
【摘要】：沙柳作为西北地区抗旱固沙树种,其特有的机制使它能够在干旱沙漠环境中生存,研究其分枝及材性发育相关遗传机制,对促进沙柳分子定向育种,培育枝条丛生型品系,增加土壤覆盖面积,防止沙土流失及利用沙柳生物质能源具有重要意义。本研究以拟南芥突变体和沙柳为植物材料,分析了拟南芥pny突变体表型变化,克隆获得了沙柳PNY基因,进行了生物信息学分析与组织特异性表达分析。主要实验结果如下：(1)研究发现拟南芥pny突变系发育迟缓,表型变化明显,分枝数量与茎生叶显著增加。拟南芥pny突变系茎解剖结构分析发现,茎基部次生木质部较野生型发达,细胞排列紧密,韧皮部发育及向外扩增较快。(2)利用同源克隆技术,在沙柳中克隆获得PNY基因,对PNY蛋白进行保守序列区域及功能结构域分析,发现该蛋白含有5个保守序列区域,2个功能结构域。进化分析表明,沙柳与杞柳和毛果杨亲缘关系较近。(3)通过qRT-PCR对PNY基因在沙柳不同组织部位进行特异性表达分析,发现PNY基因在沙柳的叶腋部位表达量最高,在茎、叶、顶芽,根中表达量依次降低。研究结果表明,PNY基因不仅影响分枝发育,也影响维管系统发育；克隆获得了沙柳PNY基因,该基因可能为转录因子；沙柳SpsPNY基因及相关植物的进化树分析表明木本植物间的PNY基因亲缘关系更近：该基因在沙柳不同组织部位的特异性表达分析表明该基因在木本植物中可能也影响维管及分枝发育。本研究成果为进一步分析PNY基因功能奠定了研究基础,对树木分子育种工作具有重要指导意义。
[Abstract]:As a drought resistant and sand fixation tree species in Northwest China, Salix willow is able to survive in arid desert environment by its unique mechanism. The genetic mechanism related to branch and wood sex development is studied to promote molecular orientation breeding of Salix, and to cultivate cluster species of branches. It is of great significance to increase soil cover area, prevent sand loss and utilize biomass energy of Salix. In this study, Arabidopsis thaliana mutants and Salix were used as plant materials to analyze the surface changes of Arabidopsis thaliana pny mutants. The PNY gene was cloned and analyzed by bioinformatics and tissue specific expression. The main results were as follows: (1) it was found that the pny mutant lines of Arabidopsis thaliana were stunted, phenotypic changes were obvious, and the number of branches and stem leaves were significantly increased. The anatomic structure analysis of pny mutant of Arabidopsis thaliana showed that the secondary xylem at the base of the stem was more developed than that of the wild type, the cells were arranged closely, and the phloem developed and expanded rapidly. (2) the PNY gene was cloned from Salix willow by homologous cloning technique. The conserved region and functional domain of PNY protein were analyzed. It was found that the protein contained 5 conserved regions and 2 functional domains. The phylogenetic analysis showed that Salix was closely related to willow and poplar. (3) the specific expression of PNY gene in different tissues of Salix willow was analyzed by qRT-PCR. It was found that the expression of PNY gene was the highest in the leaf axils of Salix willow and in the stem. The expression level of leaf, terminal bud and root decreased in turn. The results showed that the PNY gene not only affected the branching development but also the vascular system development, and cloned the PNY gene of Salix wilfordii, which may be a transcription factor. Phylogenetic tree analysis of Salix SpsPNY gene and related plants showed that the PNY gene was more closely related to woody plants. The specific expression of PNY gene in different tissues of Salix wilfordii showed that the gene might also have an effect on woody plants. Vascular and branching development. The results of this study lay a foundation for further analysis of PNY gene function and have important guiding significance for tree molecular breeding.
【学位授予单位】：内蒙古农业大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S793.9
本文编号：2402048
[Abstract]:As a drought resistant and sand fixation tree species in Northwest China, Salix willow is able to survive in arid desert environment by its unique mechanism. The genetic mechanism related to branch and wood sex development is studied to promote molecular orientation breeding of Salix, and to cultivate cluster species of branches. It is of great significance to increase soil cover area, prevent sand loss and utilize biomass energy of Salix. In this study, Arabidopsis thaliana mutants and Salix were used as plant materials to analyze the surface changes of Arabidopsis thaliana pny mutants. The PNY gene was cloned and analyzed by bioinformatics and tissue specific expression. The main results were as follows: (1) it was found that the pny mutant lines of Arabidopsis thaliana were stunted, phenotypic changes were obvious, and the number of branches and stem leaves were significantly increased. The anatomic structure analysis of pny mutant of Arabidopsis thaliana showed that the secondary xylem at the base of the stem was more developed than that of the wild type, the cells were arranged closely, and the phloem developed and expanded rapidly. (2) the PNY gene was cloned from Salix willow by homologous cloning technique. The conserved region and functional domain of PNY protein were analyzed. It was found that the protein contained 5 conserved regions and 2 functional domains. The phylogenetic analysis showed that Salix was closely related to willow and poplar. (3) the specific expression of PNY gene in different tissues of Salix willow was analyzed by qRT-PCR. It was found that the expression of PNY gene was the highest in the leaf axils of Salix willow and in the stem. The expression level of leaf, terminal bud and root decreased in turn. The results showed that the PNY gene not only affected the branching development but also the vascular system development, and cloned the PNY gene of Salix wilfordii, which may be a transcription factor. Phylogenetic tree analysis of Salix SpsPNY gene and related plants showed that the PNY gene was more closely related to woody plants. The specific expression of PNY gene in different tissues of Salix wilfordii showed that the gene might also have an effect on woody plants. Vascular and branching development. The results of this study lay a foundation for further analysis of PNY gene function and have important guiding significance for tree molecular breeding.
【学位授予单位】：内蒙古农业大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S793.9
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1 王文娟;拟南芥pny突变体分析及沙柳PNY基因克隆与组织特异性表达分析[D];内蒙古农业大学;2016年
，本文编号：2402048
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