摘要:施用赤霉素是生产上常见获取无核葡萄手段之一，但目前有关赤霉素诱导核退化分子机制尚不清楚。本研究以‘白罗莎’为试材，通过对VvPAL、VvCAD、Vv4CL基因序列结构分析发现，VvPAL、VvCAD、Vv4CL基因与木本类植物同源性均在85%以上，说明三个基因保守性较高。利用qRT-PCR技术分析了PAL、CAD、4CL在葡萄果核发育不同时期的时空表达及其应答赤霉素的模式，结果显示，木质素合成关键酶基因PAL、CAD、4CL在葡萄果核发育四个关键时期均受到赤霉素信号影响，且其表达被赤霉素抑制。本研究从分子水平认识了果核退化过程中赤霉素信号对木质素合成关键酶基因表达的影响，为进一步研究葡萄中赤霉素诱导葡萄果核退化的分子机制提供了理论依据。43585

毕业论文关键词：葡萄；木质素；GA；生物信息学；qRT-PCR；

The analysis of the expression concerning ligin in Gibberellin induces ‘White Rosa’ grape stone degradation

Abstract：The application of gibberellin is one of the most common means of obtaining seedless grapes in production. However, the molecular mechanism underlying gibberellin induced nuclear degeneration remains unknown. By analyzing the structure of the VvPAL, VvCAD, Vv4CL gene sequence and using the "White Rosa" as the test material, I found that VvPAL, VvCAD, Vv4CL gene with woody plant homology were more than 85%, indicating that the three genes are highly conserved.The spatial and temporal expression of PAL, CAD and 4CL in different stages of grapevine development was analyzed by qRT-PCR and the response model of gibberellin was analyzed.The results showed that the key enzymes of lignin synthesis, PAL, CAD and 4CL were affected by gibberellin signal in four critical periods and inhibit its expression. In this study, we investigated the effect of gibberellin signal on the expression of key enzymes in lignin synthesis from the molecular mechanism, and provided the basis for further utilization of gibberellin-induced nuclear degradation mechanism in grape.

Key words: Grape；Lignin；GA；Bio-information；qRT-PCR；

目  录

关键词 1

Abstract 1

Key words 2

1材料与方法 2

1.1 实验材料 2

1.1.1材料选择与处理 2

1.1.2基因组数据库序列来源 3

1.1.3主要仪器设备 3

1.1.4主要试剂 3

1.1.5生物信息学分析工具 3

1.1.6本地生物信息学分析软件 3

1.2 葡萄PAL、CAD和4CL生物信息学分析方法 3

1.2.1 葡萄PAL、CAD、4CL基因结构分析 3

1.2.2开放阅读框（ORF）分析与翻译 3

1.2.3基因同源性分析 3

1.2.4基因进化树的构建及蛋白功能结构域分析 3

1.3 葡萄PAL、CAD和4CL功能验证及荧光定量分析 4

1.3.1葡萄核区RNA的提取 4

1.3.2 葡萄cDNA合成 4

1.3.3 PAL、CAD和4CL基因的定量表达分析 5

1.4 葡萄硬核发育关键时期生理切片 5

2结果与分析 6

2.1葡萄硬核发育关键时期的果实横切图