摘要研究植物器官尺寸大小不仅有助于进一步揭示调控器官大小的基因网络,而且对植物形态外观、生存繁殖、胁迫抗性等品质的遗传有重要指导意义,在生产实践中有广泛的应用前景。豆科植物百脉根的BIO ORGANS基因(LjBIO)已经被证实与花器官对称性和大小调控有关。目前尚未有关于LjBIO基因具体功能和调控机制的进一步研究报告。本研究克隆了模式植物拟南芥中的 BIO ORGANS同源基因At3g24150与At4g32295,构建了其过表达载体;利用大岩桐作为研究材料,优化了大岩桐基因转化和再生体系,并进行异源基因转化,期待通过观察和测定,探索LjBIO及同源基因对花型大小颜色的潜在作用和市场应用前景。44136
毕业论文关键词:BIO ORGANS;基因转化;大岩桐;器官大小;功能研究
Abstract
Studies on plant organ size help us understand the network of size control genes, and help us to manipulate the shape, reproduction rate and stress tolerance of plants. BIO ORGANS (BIO) gene was identified to play a role in dorsoventral differentiation, organ internal asymmetry and floral organ size control in Lotus japonicus. There are few reports of its function and applications at present. In this project, we cloned the homologous genes of LjBIO in Arabidopsis, used Sinningia speciosa as materials, refined the transformation system and planed to transform these genes into it. We hope to study their function and mechanism in controlling organ size and internal asymmetry, and make use of its function in changing flower shape and size.
Keyword: BIO ORGANS; gene transformation; Sinningia speciosa; organ size; function research
目 录
摘 要 3
Abstract 3
1 前 言 5
1.1 百脉根BIO ORGANS(BIO)基因研究进展 5
1.2 BIO同源基因转化大岩桐 6
2 材料试剂及实验方法 6
2.1 材料与试剂 6
2.1.1 材料 6
2.1.2 试剂 6
2.1.3 主要仪器 6
2.2 实验方法 7
2.2.1 基因克隆 7
2.2.1.1 拟南芥cDNA获取 7
2.2.1.2 T-A克隆及测序验证 7
2.2.2 过表达载体构建 7
2.2.3 大岩桐再生体系筛选压的确定 7
2.2.4 大岩桐基因转化 7
2.2.4.1 侵染液的制备 7
2.2.4.2 大岩桐的侵染 8
2.2.4.3 转基因大岩桐表型和花发育观察 8
3 实验结果与分析 8
3.1 拟南芥BIO同源基因克隆和过表达载体构建 8
3.1.1 特异性基因扩增 8
3.1.2 表达载体的酶切验证 9
3.2 大岩桐再生体系筛选压的确定 10
3.3 大岩桐的侵染 10
3.4 大岩桐的生长与再生 11
3.5 讨论 11
3.6 展望 12
致 谢