Developmental programming of the Brassica napus seed

油菜(Brassica Napus)和从这种植物中提取的种子对加拿大经济来说是一个数十亿美元的产业，在世界各地的需求都在增加。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了种子如何以及何时发育。油菜种子可分为合子区和母体区。母体区域可进一步划分为几个亚区，包括索状体、合点点种皮、合点点增殖组织和远端种皮。我们对母体种子亚区缺乏了解，主要是因为它们不能在种子内进行分子和基因组研究。在油菜中，哪些基因负责母体分区域的发育？哪些母系次区域转录因子对种子发育至关重要？这些基因又是如何调控的呢？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，以描绘特定植物细胞类型中的基因活动模式。我们的实验室在激光显微解剖组织的加工、RNA测序库的建立以及用于分析大规模数据集的生物信息学管道的分析和开发方面积累了专业知识。我们将重点放在母系亚区的RNA上，因为我们在这些分子的分析方面已经形成了深厚的专业知识。除了缺乏具有异常种子表型的转录调控因子的转化体外，分子数据还将补充对母体亚区的深入组织学分析。这项假设驱动的研究将确定对种子发育至关重要的基因，这些基因特别在母体亚区活跃。我们将进一步研究这些调控途径是如何通过microRNAs的图谱来控制的。总的来说，NSERC发现基金将促进实验室中三名博士生、一名硕士学生和本科生的培训，并将研究母体次区域转录调节因子在种子发育中的功能作用，包括转录因子和在经济和农业上具有重要意义的油籽油菜籽中的微型RNA群体。这一开创性研究的结果将有助于那些对作物改良感兴趣的人理解种子发育。

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.