油菜转录因子PHL2调控种子含油量的功能及机制研究

油菜种子含油量是多基因调控的重要性状，提高种子含油量是重要育种目标。PHL2属于MYB-CC家族转录因子。前人研究表明，PHL2参与调控磷饥饿诱导基因的表达，进而调节植物对磷吸收与利用。我们前期研究发现，过表达油菜及拟南芥PHL2使转基因拟南芥种子总脂肪酸含量增加14.99%-31.11%。本课题拟研究油菜PHL2对种子含油量的影响及作用机制。主要研究内容包括克隆甘蓝型油菜PHL2所有拷贝，阐明其表达规律；在油菜中过表达或敲除PHL2，研究其对种子含油量的影响；利用RNA-seq研究PHL2过表达和敲除对种子发育过程中不同基因表达的影响；利用CUT&Tag、酵母单杂交、EMSA和荧光素酶等方法确定其下游靶基因；利用生物信息学、转基因及基因编辑技术确定相关重要靶基因的功能；构建PHL2对种子含油量影响的调控网络。该研究将发现PHL2在种子油脂合成调控中的新作用及机理，对全面构建油菜油脂合

Seed oil content is an important characteristic of oilseed rape, which is controlled by multigenes. The increase in seed oil content is an important target in breeding. PHL2 is a transcription factor that belongs to the MYB-CC family. Previous studies have found that PHL2 is involved in regulating the expression of phosphorus starvation-inducing genes, which regulate the uptake and utilization of phosphorus in plants. In our previous work, we found that overexpression of PHL2 from Brassica napus and Arabidopsis can increase the accumulation of total fatty acids in transgenic Arabidopsis seeds by 14.99%-31.11%. Therefore in this project we try to study the effect of PHL2 from oilseed rape on seed oil content and its mechanism. The main research contents include following aspects: cloning all copies of Brassica napus PHL2 and detecting their expression profiles during seed development; over-expressing or knocking out of PHL2 genes in Brassica napus to see the effects of PHL2 on the seed oil content, identifying the downstream target genes regulated by PHL2 with CUT & Tag, yeast one-hybrid technology, EMSA and luciferase methods; confirming the functions of PHL2-target genes related to oil synthesis by bioinformatics and transgenic and gene editing technologies; and finally constructing the regulatory network of PHL2 on seed oil content. This study will discover the new function and mechanism of PHL2 in the regulation of seed oil synthesis, which is of great significance for the comprehensive construction of the regulation network of oil synthesis in Brassica napus and the further development of new high-oil content rapeseed germplasm.