杨树环化酶基因Potri.006G237100功能分析

发布时间：2018-05-01 11:15

本文选题：毛果杨 + Portri.006G237100　；参考：《东北林业大学》2016年硕士论文

【摘要】：环化酶是一种催化核苷三磷酸生成环核苷酸的酶,广泛存在于动物、植物、微生物。环化酶编码基因呈现基因家族特征,在植物中存在多种类型,参与植物的多种生物学功能。研究报道,水稻的一类环化酶通过控制活性氧水平参与逆境胁迫。番茄红素ε-环化酶基因过量表达与光合作用有关,番茄红素β-环化酶基因参与小麦β-胡萝卜素生物合成。丙二烯环化酶基因参与作物的耐盐性,过量表达该基因增加水稻在高盐度环境下生物量,这种功能在小麦中也有类似报道。然而,环化酶基因在树木中的研究信息或功能,却报道甚少。在前期研究中我们分离一个杨树环化酶基因Potri.006G237100,它可能参与木材形成。为了鉴定该基因的功能,取得的主要研究结果如下：毛果杨环化酶基因Potri.006G237100编码区全长CDS序列含507个核苷酸,编码169个氨基酸。同源性检索显示,拟南芥、大豆及葡萄等不同植物物种均存在其同源基因。序列对比显示,该基因编码蛋白的氨基酸在不同的植物物种中存在较高的保守性。半定量RT-PCR分析结果显示,毛果杨环化酶基因Potri.006G237100在木质部、叶柄、根组织中高丰度表达,且其转录水平与茎木质化程度同步。构建Potri.006G237100-GFP融合的植物载体,在拟南芥中过量表达Potri.006G237100-GFP,用激光共聚焦显微镜检测转基因拟南芥根中GFP荧光信号,结果显示信号集中细胞质内,在周质区域也有少量信号分布,表明Potri.006G237100蛋白可能定位在细胞质。通过转化拟南芥获得Potri.006G237100过量表达遗传材料,组织解剖学结合化学染色分析显示,与野生型相比过表达Potri.006G237100基因拟南芥材料出现木质部细胞不规整、束间细胞壁增厚；过表达Potri.006G237100基因拟南芥材料的根长与野生型拟南芥根长相比变长。这些结果表明环化酶基因Potri.006G237100可能参与杨树木材(次生细胞壁)形成。
[Abstract]:Cyclase is an enzyme that catalyzes the formation of cyclic nucleotides by nucleoside triphosphate, which is widely found in animals, plants and microorganisms. Cyclase coding genes show the characteristics of gene family, and there are many types in plants, which participate in many biological functions of plants. It is reported that a class of cyclase in rice participates in stress stress by controlling the level of reactive oxygen species (Ros). The overexpression of lycopene 蔚 -cyclase gene is related to photosynthesis, and lycopene 尾 -cyclase gene is involved in 尾 -carotene biosynthesis in wheat. Allylene cyclase gene is involved in the salt tolerance of crops. Overexpression of the gene increases the biomass of rice under high salinity. This function is also reported in wheat. However, the research information or function of cyclase gene in trees is rarely reported. In previous studies, we isolated a poplar cyclase gene Potri.006G237100, which may be involved in wood formation. In order to identify the function of the gene, the main results are as follows: the full-length CDS sequence of the Potri.006G237100 coding region of poplar cyclase gene contains 507 nucleotides and encodes 169 amino acids. Homology search showed that homologous genes existed in Arabidopsis thaliana, soybean and grape. Sequence comparison showed that the amino acids encoded by the gene were highly conserved in different plant species. The results of semi-quantitative RT-PCR analysis showed that poplar cyclase gene Potri.006G237100 was highly abundant in xylem, petiole and root, and its transcription level was synchronized with that of stem lignification. The plant vector of Potri.006G237100-GFP fusion was constructed, and Potri.006G237100-GFPwas overexpressed in Arabidopsis thaliana. The fluorescence signal of GFP in transgenic Arabidopsis root was detected by laser confocal microscope. The results showed that the signal was concentrated in cytoplasm and distributed in periplasmic region. It was suggested that Potri.006G237100 protein might be located in cytoplasm. In Arabidopsis thaliana (Arabidopsis thaliana), Potri.006G237100 over-expressed genetic materials were obtained. The results of histological anatomy and chemical staining showed that compared with wild-type Arabidopsis thaliana with overexpression of Potri.006G237100 gene, the xylem cells were irregular and the cell wall between bundles was thicker than that of wild-type Arabidopsis. The root length of Arabidopsis thaliana with overexpression of Potri.006G237100 gene was longer than that of wild type Arabidopsis thaliana. These results suggest that cyclase gene Potri.006G237100 may be involved in the formation of poplar wood (secondary cell wall).
【学位授予单位】：东北林业大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S792.11;Q943.2

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