基于双基因聚合调控的月季花器官发育分子机制研究

The transgenic China Rose(Rosa chinensis), with enhanced cold-tolerance and higher growth rate in contrast to the nontransgenic ones by transforming with a dehydration- responsive element binding protein gene cloned from Medicago truncatula, MtDREB1C, and a xyloglucan endotransglycosylase gene from R. chinensis, RcXET, showed sepal-like pistils while flowering in flask. While this phenomenon was rarely found in nontransgenic plants in flask. Advanced research by high-throughput sequencing showed some flower development-related genes were expressed by the pyramided genes and two of them were closely linked with floral organdevelopment. We presume that some down-regulated genes by the pyramided genes controlled the floral organ development and parts of them changed the way of floral organ development. The CRISPR/Cas9 technology is proposed to be applied using the two genes by gene engineering in China Rose, to generate some mutants by Cas9-based gene editing. To construct the links between mutant genes and ornamental traits. Hence, this project will be significantly meaningful for illustrating the discovering useful gene resources, and even improving molecule design for breeding woody ornamentals.

耐寒相关基因MtDREB1C和生长相关基因RcXET聚合转化中国月季，获得耐寒性和生长势明显增强的转化植株，而且转化植株有组培瓶内开花和雌蕊花萼化的现象。通过转录组测序分析发现，差异基因中有一批花发育相关基因表达，其中2个与花器官发育密切相关。据此推测，聚合基因表达时，调控下游花发育基因，改变了花期和花器官的发育。拟将2个与花器官发育最为密切的基因序列，构建CRISPR/Cas9系统，转化月季，获得转化植株；通过对转化植株观赏特征的持续观察，和对转入基因跟踪测序，检测目的基因被Cas9蛋白编辑的情况，构建突变基因与观赏特征的关联模型。本研究对发现可利用基因资源、改良具复杂花器官的木本观赏植物分子育种设计，有深远意义。

前期研究中通过转录组测序分析发现，差异基因中有一批月季花发育相关基因表达，其中2个与花器官发育密切相关。本研究将2个与花器官发育最为密切的基因序列，构建CRISPR/Cas9系统，转化拟南芥，筛选到荧光果荚；CRISPR-cas9-RcAP2转化月季，获得荧光转化芽；为后续观赏特征的观察、和基因测序，检测目的基因被Cas9蛋白编辑的情况，构建突变基因与观赏特征的关联模型奠定了基础。本研究对发现可利用基因资源、改良具复杂花器官的木本观赏植物分子育种设计，有深远意义。