牡丹花粉管与雌蕊识别过程的转录组分析及调控结实关键基因挖掘

结实是限制油用牡丹产量和种质创新的重要因素，受精成败又是影响牡丹结实性的关键生殖过程，此过程涉及到很多基因，是一个非常复杂而精细的生命现象，然而牡丹中有关这一过程的内在机制却一无所知，严重影响了牡丹的开发和利用，亟待开展相关研究。本项目组前期通过结实性调查、细胞学观察，对牡丹受精过程中的受精障碍进行了研究。本项目拟在前期研究的基础上，通过进一步对受精过程的细胞学观察和结实性调查，结合高通量测序技术，以授粉前、授粉后不同时期的雌蕊为材料，获得授粉受精过程中的转录谱，筛选可能参与牡丹花粉管-雌蕊识别的关键基因；对候选基因通过荧光定量PCR技术进行表达分析；并通过过表达拟南芥和牡丹体内的基因沉默技术对候选基因的功能进行鉴定，从细胞学和分子生物学角度揭示牡丹受精过程中花粉管-雌蕊识别的分子调控机制，为提高牡丹结实率、指导种子生产，提升远缘杂交育种效率、种质创新奠定基础。

Seed setting is an important factor limiting tree peony seed oil yield and germplasm innovation, while, fertilization success is a key factor for tree peony seed setting. Multiple genes involved in fertilization process, which is a very complicate and delicate biological phenomena, however, the molecular mechanism on this process of tree peony is still unknown, which seriously affected the development and utilization of tree peony, therefore, it is urgent to carry out related research. We had analyzed fertilization barrier in the process of tree peony fertilization by seed setting investigation and cytological observation. Based on our previous study, we will attempt to obtain the transcript profiling during the process of fertilization of tree peony that using pistils before or after pollination as materials, through next-generation sequencing technique, and analyze the differential expression profile, try to screen the key genes which may be responsible for the recognition between pollen tube and pistil. Meanwhile, the expression level of candidate genes will be validated by qRT-PCR. Furtherly, the gene silencing, transforming to model plants and functional complementation methods were also used to investigate the candidate genes biological function. Finally, we will demonstrate the mechanism of recognition between pollen tube and pistil in fertilization process by combining the results of cytological and molecular biology, which will lay the foundation for understanding the process of fertilization, improving the seed setting rate for tree peony seed production and increasing the efficiency of breeding for germplasm innovation.