摘要植物器官大小是由一系列基因网络所调控，这一现象的研究对植物形态外观、生存繁殖等都具有遗传上的指导意义，在生产实践中也具备相当可观的应用前景。百脉根的BIO ORGANS基因（LjBIO）已经被证实与花器官的大小调控密切相关。但目前的研究关于LjBIO基因功能的具体报告少之又少，也未见对拟南芥中同源基因At3g24150和At4g32295的功能揭示。在本研究中，我们利用模式生物拟南芥为研究材料，克隆研究BIO ORGANS同源基因，通过基因干涉的原理构建其基因干涉载体，用于转化拟南芥并限制其表达量，探索其在调节花器官大小、花色中的生物学功能。44208

毕业论文关键词：BIO ORGANS基因；拟南芥；器官大小；RNA干涉；功能研究

Abstract

Plant organ size is regulated by a series of gene networks. Study of this phenomenon is instructive to the appearance of plant morphology, survival and reproduction. It has considerable prospects in the production. It is verified that BIO ORGANS Gene (LjBIO) is closely related to the regulation of floral organ size. However, there are very few studies reporting the specific gene function of LjBIO, There is no report about its homolog genes At3g24150 and At4g32295 in Arabidopsis. In this work, we used the model plant Arabidopsis as the materials, cloned the LjBIO homologous gene, constructed their gene-interference vectors, in order to decrease gene expression level with transformation of Arabidopsis, and explore their potential biological function in flower colour and size controll.

Keyword: BIO ORGANS gene; Arabidopsis thaliana; organ size; RNAi; function research

目   录

1 前言 5

2 BIO同源基因的克隆和干涉载体构建 5

2.1 实验材料、试剂与设备 5

2.1.1 实验材料 5

2.1.2 试剂 6

2.1.3 实验用仪器 6

2.2 实验方法 6

2.2.1 基因克隆 6

2.2.2 干涉片段的克隆 9

2.2.3 干涉载体的构建 10

2.2.4 干涉载体转化农杆菌 10

2.3 结果和分析 11

2.3.1 At4g32295.1、At4g32295.2和At3g24150基因克隆 11

2.3.2 I-At4g32295和I-At3g24150干涉片段克隆 12

2.3.3 At4g32295-RNAi以及At3g-24150RNAi干涉载体构建 14

2.3.4 干涉载体转化农杆菌 15

3 BIO 同源基因干涉载体转基因拟南芥的生长以及表型分析 16

3.1 材料以及相关试剂 16

3.1.1 实验材料及药品 16

3.1.2 实验仪器 16

3.2 实验方法 16

3.2.1 拟南芥培养 16

3.2.2 拟南芥基因转化与筛选 16

3.2.3 转基因植株的分子鉴定 17

3.3 结果与分析 17

3.3.1拟南芥At4g32295.1、At4g32295.2、At3g24150基因组织特异性表达 17

3.3.2拟南芥转基因植株的分子鉴定 17

3.3.3 讨论