摘要：荷花是中国十大传统名花之一，具有很高的欣赏、经济和文化价值。在栽培中发现荷花在每个立叶基部都会形成花芽，但是荷花开花的数量通常比立叶的数目少的多。弱光下花芽的败育可能是荷花开花数量远少于立叶的数目的主要原因。前人研究表明MADS-box基因家族在植物开花过程的调节和控制中起关键作用。本研究通过对荷花的MADS-box基因进行全基因组范围鉴定，共鉴定到50个MADS-box家族成员。对获得的荷花MADS-box基因与拟南芥和水稻的MADS-box基因构建系统发育树，可划分为MIKC（25个基因），MIKC*（1个基因），Mα（14个基因），Mβ（4个基因），Mγ（6个基因）5个亚家族。组织表达量分析显示M型MADS-box基因亚家族中，除M-β亚群外，其他在大多数组织中表现出极低的水平。MADS-box基因表现出与拟南芥或其他植物物种中类似的表达模式，但也存在许多具有独特表达谱的基因。研究进一步通过高通量测序技术分析荷花品种“红晕蝶影”在遮阴处理下的转录组变化，获得6个显著响应弱光处理的MADS-box基因。这些基因可能在荷花响应遮阴调控花芽败育中发挥重要的功能，在未来通过转基因手段进行进一步功能鉴定将有助于揭示其花芽败育的分子机理。
关键词：荷花；MADS-box基因；花器官发育
Identification of MADS-box Gene in Lotus and Its Function Analysis in Flower Bud Development
Abstract：Lotus is one of the top ten traditional flowers in China, with high ornamental, economic and cultural values. It was observed that lotus often produce a flower bud along side leaf. However, the flower number often far less than number of leaves, which indicated that flower bud abortion might the main cause of flower number reduction under lower light condition. Previous work showed that MADS-box genes might play an important role in flowering regulation. In this work, 50 lotus MADS-box (NnMADS-box) genes were identified in a genome-wide screen. Taken with Arabidopsis and rice MADS-box genes, a phylogenetic tree was constructed with the predicted amino acid sequence of genes. The phylogenetic tree was clearly pided into five subfamilies: MIKC (25 genes)，MIKC *(1 gene)，Mα (14 genes)，Mβ (4 genes)，Mγ (6 genes). Expression patterns in different lotus tissues showed that M-type subfamily genes showed extremely low level in most tissues except M-beta subgroup. Whereas many of the MADS-like genes identified in lotus exhibit expression patterns similar to those in Arabidopsis or other plant species, there are also a number of genes with unique expression profiles. We further performed a high-throughput gene expression analysis of lotus upon shade treatment, and 6 MADS-box genes demonstrated significant changes, and could be candidate genes in lotus shade response. Further characterization and the generation of transgenic plants containing these genes could help to elucidate its function.
Key words: lotus; MADS-box gene; flower bud development
目 录
摘要3
关键词…3
Abstract…3
Key words 3
1 前言 4
1.1 MADS-box基因家族概述4
1.2 MADS-box家族的基因结构及分类…4
1.3 研究的目的和意义4
2 材料方法 5
2.1鉴定MADS-box基因5
2.2蛋白质特性分析5
2.3引物设计与PCR扩增5
2.4构建系统发育树…5
2.5转录组数据分析…5
2.6 植物材料处理，RNA提取和文库构建准备以及测序…5
3结果与分析6
3.1 鉴定荷花MADS-box基因6
3.2 荷花MADS-box家族基因的系统发育分析7
3.3 MADS-box基因表达分析10
3.3.1 遮阴处理下荷花转录组及MADS-box 基因分析 13 荷花MADS-box基因鉴定及其在花芽发育中的功能分析:http://www.chuibin.com/shengwu/lunwen_206221.html