Expression analysis of root cell types after treatment with sulfur deficient media

Cell-type specific transcriptional profiles were generated by FACS (Fluorescence Activated Cell Sorting) sorting of roots that express cell-type specific GFP-reporters. Five different GFP-reporter lines were used. FACS cell populations were isolated from roots grown under sulfur deficient conditions for 3 hours. Stress responses in plants are tightly coordinated with developmental processes, but the interaction between these pathways is poorly understood. Here we use genome-wide assays at high spatial and temporal resolution to understand the processes that lnk development and stress in the Arabidopsis root. Our meta-analysis finds little evidence for a universal stress response. Common stress responses appear to exists and, analagous to animal systems, many of them show cell-type specificity, suggesting a convergent evolutionary theme in multicellular organisms. Common stress responses may be mediated by cell identity regulators, as mutations in these genes resulted in altered responses to stress. Our results reveal surprising linkages between stress and development at cellular resolution, and show the power of multiple genome-wide datasets to elucidate biological processes. 3 replicates for each of 5 cell types for sulfur deficient media

本研究通过荧光激活细胞分选术（Fluorescence Activated Cell Sorting，FACS）对表达细胞类型特异性绿色荧光蛋白报告基因的拟南芥根系进行分选，制备了细胞类型特异性转录组图谱。本研究共使用5种不同的GFP报告基因株系，所分离的FACS细胞群体均来自于在硫缺乏条件下培养3小时的根系。植物的胁迫响应与发育进程紧密协同，但二者之间的调控通路互作机制仍有待深入阐释。本研究采用高时空分辨率的全基因组检测技术，解析拟南芥根系中发育与胁迫响应的关联调控过程。荟萃分析结果显示，并不存在通用的植物胁迫响应通路；尽管存在共通的胁迫响应通路，且与动物系统类似，其中多数通路均表现出细胞类型特异性，这提示多细胞生物中存在一类趋同进化的调控范式。共通胁迫响应或由细胞身份调控因子介导，此类基因的突变会导致植物胁迫响应发生改变。本研究结果在单细胞分辨率下揭示了胁迫与发育之间未曾预料的关联，并证实了多组全基因组数据集在解析生物学过程中的重要价值。硫缺乏培养基条件下的5种细胞类型均设置了3次生物学重复。