赤霉素诱导葡萄胚败育相关基因的克隆序列分析与时空表达特征鉴定
时间:2018-09-18 20:45 来源:毕业论文 作者:毕业论文 点击:次
| 摘要：本实验利用赤霉素处理‘白罗莎’葡萄（Rosario Bianco）诱导其胚败育的体系，对胚败育相关的VvAGL15、VvAGL23、VvLEC1基因开展研究。通过RT-PCR克隆获得VvAGL15、VvAGL23、VvLEC1基因的编码区序列，长度分别为744bp、690bp、636bp。同源性分析表明，其与其它物种同源基因具有较高的同源性，且与拟南芥具有相同的功能结构域；启动子作用元件分析发现这三个基因均有胚乳组织特异发育相关的作用元件，推测其可能与种胚发育相关；VvAGL15、VvAGL23基因有赤霉素响应作用元件表明其可能响应赤霉素信号。qRT-PCR时空表达分析显示它们在葡萄果实发育的不同时期存在差异表达，尤其在硬核期表达水平相对较高，且赤霉素在该时期显著下调上述基因表达，多效唑则作用相反，表明赤霉素可能通过调控这三个基因的表达从而抑制胚发育。28365 毕业论文关键词：赤霉素、葡萄胚败育、基因克隆、序列分析、时空表达分析 Cloning, sequence and expression analysis of genes related to Gibberellin induced embryo abortion in grape Abstract：In this study, gibberellin treatment to 'Rosario Bianco' was used to induce the embryo abortion system. The VvAGL15, VvAGL23 and VvLEC1 genes related to embryo abortion were studied. The coding sequence of VvAGL15, VvAGL23 and VvLEC1 genes were 744bp, 690bp and 636bp respectively by RT-PCR. Homology analysis showed that it had high homology with other species homologous genes and had the same functional domain as Arabidopsis thaliana. The promoter acting elements analysis showed that the three genes all had acting elements which were related to specific endosperm tissues development , suggesting that it may be associated with embryo development; VvAGL15 and VvAGL23 gene have a gibberellin response element indicating that it may respond to gibberellin signaling. The temporal and spatial expression analysis of qRT-PCR showed that they were differentially expressed in different stages of fruit development, especially higher in the hard nucleus, and gibberellin significantly down-regulated the gene expression in this period , the opposite acting of paclobutrazol suggested that gibberellin may inhibit the development of embryo by regulating the expression of these three genes. Key words: Gibberellin; Grape embryo abortion; gene cloning; sequence analysis; Expression analysis 目 录 摘要 1 关键词 1 Abstract 1 Key words 1 引言 1 1 材料与方法 2 1．1 生理形态观察材料与方法 2 1．1．1 材料与试剂 2 1．1．2 方法 2 1．2 目的基因编码区克隆材料与方法 2 1．2．1 材料 3 1．2．2 方法 3 1．3 目的基因序列分析网站及软件 6 1．3．1 材料 6 1．3．2 方法 7 1．4 目的基因时空表达分析材料与方法 7 1．4．1 实验材料与试剂 7 1．4．2 实验方法 7 2 结果与分析 8 2．1 生理形态观察与分析 8 2．1．1 果穗发育分析 8 2．1．2 不同处理果实果核区形态比较 9 2．2 VvAGL15、VvAGL23、VvLEC1基因编码区克隆和载体构建 9 2．2. 1 葡萄果实总RNA提取和cDNA的合成 9 2．2. 2 目的基因编码区克隆 10 2．2. 3 目的基因序列分析 10 (责任编辑：qin) |