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摘要拟南芥是植物界中的果蝇、分子生物学研究的模式物种，拥有生长周期短、种子量大、突变体丰富、遗传背景清楚、基因组小等优点，在花卉基因工程中得到了广泛的应用。随着转基因植物研究的逐步展开，依赖农杆菌介导的转基因技术，拟南芥成为植物遗传转化的良好材料并被普遍用于研究基因功能机制。48010
随着现代生物技术的发展，越来越多与花器官发育相关的基因得到克隆与研究。被证实与花器官对称性和大小调控有关的百脉根BIO ORGANS（BIO）基因，其具体功能和调控机制尚不清晰，也未见任何其同源基因的研究报告。本研究选择以模式植物拟南芥作为研究对象，克隆拟南芥中BIO同源基因At4g32295和At3g24150，并构建相应的过表达载体，探索其在调控花器官表型变化中的具体生物学功能，现阶段取得的成果如下：
1、克隆了拟南芥BIO的同源基因At4g32295.1、At4g32295.2和At3g24150，构建了过表达载体35S:At4g32295.1、35S:At4g32295.2和35S:At3g24150。
2、对野生型拟南芥进行半定量RT-PCR鉴定，明确了At4g32295.1、At4g32295.2和At3g24150这3个BIO同源基因的时空表达模式。在2周大的拟南芥幼苗中，三个基因的表达量表现为地上部分强于地下部分；5周开花的成苗，其茎中的表达量低于其他组织和器官，三个基因在花序、茎生叶、莲座叶中的表达量不一致。以上现象说明三者之间可能存在不一样的调控机制。
3、用构建的过表达载体转化拟南芥，已经得到了6个35S:At4g32295.1、8个35S:At4g32295.2、7个35S:At3g24150的转基因纯合植株。
4、通过对表型的观察发现，一个完整的植株世代中，转基因过量系与野生型植株间无明显差异。
毕业论文关键词：拟南芥； BIO ORGANS基因； 功能研究； 器官大小
Abstract
Arabidopsis thaliana in the plant kingdom is as the flies in the animal kingdom. It’s a model plant for molecular biology study, which has short growth period, large quantity seeds, rich mutants, clear genetic background, and a relatively small genome. It has been widely used in the flower genetic engineering. With the academic research of transgenic plants gradually unfolds, depends on the agrobacterium mediated gene transformation, Arabidopsis thaliana is becoming a good material for studying gene function and mechanism.
With the development of modern biotechnology, more and more floral genes have been cloned and studied. BIO ORGANS gene from Lotus japonicus was reported to be involved in the size regulation of flower organ size and symmetry, however its function and regulatory mechanism is not clear yet. No studies of its homologous gene in other species has been reported. In this study, we chose Arabidopsis thaliana as a model,cloned the BIO homologous gene At4g32295 and At3g24150, constructed their over-expression vectors, in order to explore their biological functions. At the present stage achievements are as follows:
1. BIO homologous genes At4g32295. 1, At4g32295. 2 and At3g24150 in Arabidopsis were cloned, the over-expression vectors of 35s:At4g32295.1, 35s:At4g32295.2 and 35s:At3g24150 were constructed.
2. Semi-quantitative RT-PCR of these BIO homologous genes was used to detect their expression pattern in the wild type Arabidopsis thaliana. In 2-week-old seedlings, the expression level was lower in roots than in shoots for all of them; In 5-week-old flowering plants, the expression level was the lowest in stem, but was varied in inflorescence, cauline leaves and rosette leaves .
3. Homozygous transgenic plants of 6 35S:At4g32295.1 lines, 8 35S:At4g32295.2 lines and 7 35S:At3g24150 lines were obtained.
4. Through the observation of phenotypes during one whole generation, no obvious differences were found between the transgenic plants and wild type.
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