Single-nucleus sequencing deciphers lineage trajectories in rice pistils [snRNA-seq]

Angiosperms possess a characteristic life cycle with an alternation of sporophyte and gametophyte generations, which happens in plant organs like pistils. Rice pistils contain ovules and receive pollen for successful fertilization to produce grains. The cellular expression profile in rice pistils is largely unknown. Here we show a cell atlas of rice pistils before fertilization by droplet-based single-nucleus RNA sequencing. The Ab initio marker identification validated by in situ hybridization assists the cell-type annotation, revealing cell heterogeneity between ovule and carpel lineages. Overall design: Perform snRNA-seq for 1N (haploid; gametophyte) and 2N (diploid; sporophyte) nuclei and bulk RNA-seq for nuclei of pistils and whole cells of upper and lower part of pistils

被子植物(Angiosperms)具有典型的孢子体(sporophyte)与配子体(gametophyte)世代交替的生命周期，该过程发生在雌蕊(pistil)等植物器官中。水稻雌蕊包含胚珠，可接收花粉以完成受精并产生籽粒。目前人们对水稻雌蕊的细胞表达谱尚缺乏充分认知。本研究通过液滴法单细胞核RNA测序(single-nucleus RNA sequencing, snRNA-seq)，构建了受精前水稻雌蕊的细胞图谱。通过原位杂交(in situ hybridization)验证的从头标记基因鉴定，助力了细胞类型注释，揭示了胚珠与心皮(carpel)谱系间的细胞异质性。总体实验设计：对单倍体(1N；配子体)和二倍体(2N；孢子体)细胞核开展单细胞核RNA测序，并对雌蕊细胞核以及雌蕊上下部位的完整细胞进行批量RNA测序(bulk RNA-seq)。