Developmental programming of the Brassica napus seed

油菜籽和从这种植物中提取的种子代表了加拿大经济中数十亿美元的产业，并且在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，它规定了种子如何以及何时发育。油菜籽可分为受精卵区和母系区。母区可进一步分为几个亚区，包括束、合点种皮、合点增殖组织和远端种皮。我们对母系种子子区域的了解是缺乏的，主要是因为它们无法在种子内进行分子和基因组研究。油菜籽母系分区发育的基因是什么？哪些母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解剖技术结合下一代RNA测序来研究这些问题，以分析特定植物细胞类型的基因活性模式。我们的实验室在激光显微解剖组织的处理、RNA测序文库的建立以及分析和开发生物信息学管道以分析大规模数据集方面拥有丰富的专业知识。我们专注于母体亚区的rna，因为我们在分析这些分子方面已经发展了深厚的专业知识。分子数据将与深入的母体亚区组织学分析以及缺乏转录调节因子的异常种子表型的转化子相补充。这一假设驱动的研究将确定对种子发育至关重要的基因，这些基因在母系亚区特别活跃。我们将进一步研究如何通过microrna的分析来控制这些调控途径。总的来说，这项NSERC发现基金将促进三名博士生、一名硕士生和实验室本科生的培训，并将研究母亲次区域转录调控因子在种子发育中的功能作用，包括转录因子和微RNA群体，在经济和农业上都很重要的油籽，油菜籽。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.