香石竹2n配子形成与PS1基因相关机理研究

项目名称： 香石竹2n配子形成与PS1基因相关机理研究

项目编号： No.31260491

项目类型： 地区科学基金项目

立项/批准年度： 2013

项目学科： 农业科学

项目作者： 周旭红

作者单位： 云南省农业科学院

项目金额： 45万元

中文摘要： 利用2n配子进行有性多倍化，比无性多倍化具有更高的适应性和杂合性，是一种有效的多倍体育种途径。我们研究发现香石竹多为二倍体，大多数能产生低频2n配子，纺锤体异常是香石竹2n配子形成的主要原因。在拟南芥中PS1基因调控纺缍体的方向，PS1基因突变能产生高频率2n配子。研究香石竹PS1基因有助于揭示2n配子形成的分子遗传学机制，加快多倍体育种进程。本研究以产生2n配子频率不同的7个二倍体香石竹品种为研究对象，旨在获得PS1同源短片断的基础上，克隆PS1基因全长cDNA，分析其序列特征、预测蛋白质结构和功能，比对品种间序列的差异，寻找可能的突变位点；实时定量荧光PCR法研究其表达模式，分析PS1基因的表达水平与2n配子形成频率的关系；构建过量表达载体和RNAi表达载体，验证PS1基因的功能，揭示PS1基因与香石竹2n配子形成的相关性，为有效利用2n配子进行多倍体育种奠定基础。

中文关键词： 香石竹；2n配子；DcPS1；表达谱；转基因

英文摘要： Sexual polyploidization, as a result of 2n gamete formation, has higher adaptability and heterozygosity than asexual polyploidization, which is thought to be the effective way involved in the polyploidy breeding. Our research found most of the Dianthus caryophyllus cultivars were diploid which could form low frequency 2n gametes, the main cytological mechanisms for the formation of 2n pollen were abnormal spindles. Studies showed that AtPS1 (Arabidopsis thaliana Parallel Spindle 1) might directly control the orientation of spindles, and its mutants product high frequency 2n gametes. So, researching PS1 gene of D. Caryophyllus may reveal molecular genetics mechanism of 2n gamete formation and speed up the polyploidy breeding process. In this study, on the basis of obtaining PS1 homologous short fragment, a PCR-based homologous cloning strategy is used to clone the cDNA sequences of PS1 genes from the seven D. caryophyllus diploid cultivars which form different frequency of 2n gamete. The cDNA sequence characteristics are analyzed，and its protein structure and function are predicted. Sequence alignment is used between different varieties in order to find possible mutant locus. Meanwhile Real-time PCR is applied to study the gene expression and the relationship between PS1 expression level and 2n gamete formation f

英文关键词： Carnation；2n gamete；DcPS1；Expression profiling；Transgenic