百山祖冷杉（Abies beshanzuensis）是我国一级重点保护野生植物，处于极度濒危状态。本项目拟以百山祖冷杉原生树和嫁接树为材料，通过对不同发育时期的胚拯救以及生长曲线的测定，明确胚与胚乳发生败育的时间与阶段，建立长期稳定的离体保存体系；通过研究不定芽诱导及其增殖技术，创建高效的快繁体系，增加百山祖冷杉原生树种群数量；通过MSAP分析不同发育阶段胚乳（雌配子体）及胚培养后代DNA甲基化图谱，明确嫁接诱导的百山祖冷杉胚乳及其后代的表观遗传多样性；利用small RNA测序技术，明确日本冷杉对百山祖冷杉small RNA的种类和数量的影响；结合转录组测序技术，筛选胚乳发育关键基因，分析胚与胚乳发生败育的规律，阐明百山祖冷杉内部濒危机制。研究结果将对我国开展百山祖冷杉及其他濒危野生植物种质资源保存、繁育和保护工作起到引领与示范作用，为利用嫁接技术保护和繁育濒危植物提供了确凿的分子证据。

Abies beshanzuensis has been incorporated into our National Key Protected Wild Plant Species List as first-rank protected wild plant species, which has been extremely threatened. This project plans to use the wild and grafted Abies beshanzuensis for research materials. Clarifying embryo and endosperm abortion time and stage and establishing long-term and stable in vitro conservation system through embryo rescue at different developmental stages and determination of the growth curve. Creating efficient propagation technology of Abies beshanzuensis to increase the population through exploring system of induction and proliferation of adventitious buds. Clarifying epigenetic diversity of Abies beshanzuensis endosperm and its offspring through MSAP technology to analyze the DNA methylation patterns of the endosperm (female gametophyte) and progeny. Clarifying the effect of Abies firma on the type and amount of small RNA of Abies beshanzuensis through small RNA sequencing technique. Selecting key genes of endosperm development through transcriptome sequencing. Elucidating endangerment mechanism of Abies beshanzuensis through analyzing the rules of abortion of embryo and endosperm. This project will play an exemplary role in the conservation and propagation of the germplasm of Abies beshanzuensis and other endangered plants in China, and provide molecular evidences for grafting techniques in protecting and breeding threatened plants.