Developmental programming of the Brassica napus seed

油菜（Brassica napus）及其种子对加拿大经济来说是一个价值数十亿美元的产业，世界各地的需求正在增加。种子发育和生产的基础是一个复杂的调控基因和途径网络，这些基因和途径指定种子如何以及何时发育。油菜种子可分为合子区和母体区。母体区又可进一步分为珠柄、合点端种皮、合点端增殖组织和远端种皮。我们对母本种子亚区的了解是缺乏的，主要是因为它们在种子内的分子和基因组研究中不可及。哪些基因负责油菜母体亚区发育？哪些母源亚区转录因子对种子发育至关重要？这些基因是如何调控的？我们将使用激光显微切割技术结合下一代RNA测序来研究这些问题，以分析特定植物细胞类型中的基因活性模式。我们的实验室在激光显微切割组织的处理，RNA测序库的构建以及生物信息学管道的分析和开发方面积累了专业知识，以分析大规模数据集。我们专注于母体亚区的RNA，因为我们在这些分子的分析方面拥有深厚的专业知识。除了缺乏转录调节因子的异常种子表型的转化体外，还将对母体亚区进行深入的组织学分析，以补充分子数据。这一假设驱动的研究将确定种子发育所必需的基因，这些基因在母体亚区中特别活跃。我们将进一步研究这些调控途径是如何通过microRNA的分析来控制的。总的来说，这个NSERC发现补助金将促进三名博士生，一名硕士生和实验室本科生研究人员的培训，并将研究种子发育的母体亚区转录调节因子的功能作用，包括经济和农业上重要的油菜籽，油菜籽中的转录因子和微RNA群体。这项开创性研究的结果将有助于那些对作物改良感兴趣的人了解种子发育。

Canola (Brassica napus) and the seeds derived from this plant represent a multi-billion dollar industry to the Canadian economy and are increasing in demand around the world. Underlying seed development and production is a complex network of regulatory genes and pathways that specify how and when a seed develops. Seeds of canola can be divided into both zygotic and maternal regions. The maternal region can further be divided into several subregions including the funiculus, chalazal seed coat, chalazal proliferating tissue and the distal seed coat. Our understanding of maternal seed subregions is lacking mostly because of their inaccessibility within the seed for molecular and genomic studies. What are the genes responsible for maternal subregion development in canola? What maternal subregion transcription factors are essential for seed development? and How are these genes regulated? We will be studying these questions using laser microdissection technology coupled to Next Generation RNA sequencing to profile patterns of gene activity in specific plant cell types. Our lab has developed expertise in the processing of laser microdissected tissues, building of RNA sequencing libraries and the analysis and development of bioinformatics pipelines to analyze large-scale datasets. We are focusing on the RNAs of the maternal subregions because we have developed deep expertise in the analysis of these molecules. The molecular data will be complemented with in depth histological analysis of maternal subregions in addition to transformants lacking transcriptional regulators with aberrant seed phenotypes. This hypothesis driven research will identify genes essential for seed development that are active specifically in maternal subregions. We will further study how these regulatory pathways are controlled through the profiling of microRNAs. Collectively, this NSERC Discovery Grant will promote the training of three PhD students, one MSc student and undergraduate researchers in the lab and will examine the functional role of maternal subregion transcriptional regulators on seed development including transcription factor and micro RNA populations in the economically and agriculturally important oilseed, canola. The results of this seminal research will contribute to the understanding of seed development for those interested in crop improvement.