大蒜体细胞胚发生的分子调控网络构建和关键基因功能分析

大蒜作为具有重要经济价值的无性繁殖蔬菜，病毒积累和种性退化严重，遗传改良和种质创新困难。常规大蒜组培技术繁殖系数低、种苗质量差、玻璃化率高、继代培养难。体细胞胚途径能克服上述不足，然而大蒜体细胞胚发生体系尚不稳定，且分子调控机理仍为空白。在我们前期大蒜体细胞胚发生研究的基础上，本项目通过优化培养条件，建立体细胞胚稳定发生体系；通过分析体细胞胚不同阶段组织和细胞的形态与解剖特征，构建大蒜体细胞胚的精确鉴定体系；利用第三代全长转录组测序技术获得大蒜转录组水平的基因组，结合第二代测序技术获得体细胞胚发生不同阶段的特异mRNA、lncRNA和miRNA，进而构建全转录水平的分子调控网络；通过大蒜转基因等技术分析关键基因在体细胞胚发生过程中的功能，明确其调控模式。本项目研究结果不仅对丰富植物体细胞胚发生机理有重要理论意义，而且对大蒜种性恢复、脱毒快繁产业化和品种改良有重要应用价值。

Garlic is the commercial vegetable species which were propagated by the vegetative propagation. Virus accumulation and degeneration of garlic species are serious, and it is difficult for the improvement and innovation of germplasm resources. Convention tissue culture has the disadvantage of low propagation coefficiency, low quality of plantlets, high hyperhydric rate and difficult to subculture. Somatic embryogenesis can solve the problems above. However, the somatic embryogenesis system of garlic is not stable and its molecular mechanism is still unclear. Based on our previous study, culture condition was optimized to establish the stable somatic embryogenesis system. Morphological and anatomical characteristics of different tissue and cell during somatic embryogenesis were analyzed to construct precise detection system. The third-generation sequencing method of full-length transcriptome was applied to get the garlic genome of the transcriptomic level. The second-generation sequencing method was applied to get the specific mRNA, lncRNA, and miRNA under the different stage of somatic embryogenesis. The molecular regulatory network of whole RNA level will be constructed. The transgenic technology of garlic will be applied to analyze the function and regulatory mode of key genes of somatic embryogenesis. This project not only can enrich the theory of plant somatic embryogenesis but also have great application value on the recovery of garlic degeneration, the industry of virus-free and rapid propagation and genetic improvement of garlic.