植物单细胞转录组测序研究进展

doi:10.13560/j.cnki.biotech.bull.1985.2020-1011

摘要/Abstract

摘要：

单细胞转录组测序是一种在单细胞水平上研究基因表达的技术。多孔板法和液滴法是目前应用于植物研究的两类主要的单细胞转录组技术。首先概述了植物单细胞转录组测序的技术原理和数据分析流程,然后介绍了植物单细胞转录组的研究进展,重点阐述了单细胞转录组测序技术在鉴定植物细胞类型、揭示细胞演化轨迹和构建细胞间调控网络中的应用。单细胞转录组测序技术可为植物领域的研究提供新视角,有助于识别和理解复杂组织中关键的细胞生物学过程。

关键词: 单细胞, 转录组测序, 液滴法, 多孔板法, 细胞类型特异性

Abstract:

Single-cell RNA-Seq（scRNA-seq）is a technology to analyze gene expression at single cell level. Currently PCR plate-based and droplet-based protocols are 2 major popular ones used in the studies of plant. Here,we firstly outline the technical principles and data analysis procedure of plant scRNA-seq,and then introduce its research status,focusing on how scRNA-seq can be used to detect distinct plant cell types,to reveal cell evolution trajectories and to construct inter-cell regulatory networks. ScRNA-seq may provide a new perspective on plant research,and help to understand and identify critical cellular biological processes in complex tissues.

Key words: single cell, RNA-sequencing, droplet-based protocol, PCR plate-based protocol, cell-type-specific

李益, 孙超. 植物单细胞转录组测序研究进展[J]. 生物技术通报, 2021, 37(1): 60-66.

LI Yi, SUN Chao. Research Progress in Single-Cell RNA-Seq of Plant[J]. Biotechnology Bulletin, 2021, 37(1): 60-66.

图/表 3

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建库方法	分析技术	捕获细胞数目	扩增技术	转录本覆盖率	参考文献
多孔板法	Smartseq2	94-384/plate	PCR	全长	[10]
	CEL-seq2	94-384/plate	IVT	3'端	[11]
液滴法	Chromium	80 000+	PCR	3'端	[13]
	Drop-seq	50 000+	PCR	3'端	[14]
	inDrop	40 000+	PCR	3'端	[15]

建库方法	分析技术	捕获细胞数目	扩增技术	转录本覆盖率	参考文献
多孔板法	Smartseq2	94-384/plate	PCR	全长	[10]
	CEL-seq2	94-384/plate	IVT	3'端	[11]
液滴法	Chromium	80 000+	PCR	3'端	[13]
	Drop-seq	50 000+	PCR	3'端	[14]
	inDrop	40 000+	PCR	3'端	[15]

建库方法	分析技术	物种	样本类型	细胞数	总基因数（转录本数）	基因数（转录本数）/细胞	参考文献
多孔板法	Smart-seq2	拟南芥	根	238	——	——	[8]
	Cell-seq2	玉米	生殖细胞	144	101 245*	——	[9]
	Cell-seq2	拟南芥	静止中心细胞	24	——	14 000	[53]
			中柱细胞	7	——	4 312	[53]
液滴法	Chromium	拟南芥	根（3个生物学重复）	7 522	>22 000	5 000	[16]
	Chromium	拟南芥	根（2个生物学重复）	4 727	16 975	4 276	[17]
	Chromium	拟南芥	根	3 121	22 419	2 445	[18]
			根（热胁迫）	1 009	21 237	1 009	[18]
			根	1 079	22 971	4 079	[18]
	Chromium	拟南芥	根	7 695	23 161	——	[19]
	Chromium	拟南芥	子叶	12 844	——	——	[20]
	Drop-seq	拟南芥	根	6 102	——	>1 000*	[54]
			根（1%蔗糖处理）	6 096	——	——	[54]
	Drop-seq	拟南芥	根	374	——	——	[55]

建库方法	分析技术	物种	样本类型	细胞数	总基因数（转录本数）	基因数（转录本数）/细胞	参考文献
多孔板法	Smart-seq2	拟南芥	根	238	——	——	[8]
	Cell-seq2	玉米	生殖细胞	144	101 245*	——	[9]
	Cell-seq2	拟南芥	静止中心细胞	24	——	14 000	[53]
			中柱细胞	7	——	4 312	[53]
液滴法	Chromium	拟南芥	根（3个生物学重复）	7 522	>22 000	5 000	[16]
	Chromium	拟南芥	根（2个生物学重复）	4 727	16 975	4 276	[17]
	Chromium	拟南芥	根	3 121	22 419	2 445	[18]
			根（热胁迫）	1 009	21 237	1 009	[18]
			根	1 079	22 971	4 079	[18]
	Chromium	拟南芥	根	7 695	23 161	——	[19]
	Chromium	拟南芥	子叶	12 844	——	——	[20]
	Drop-seq	拟南芥	根	6 102	——	>1 000*	[54]
			根（1%蔗糖处理）	6 096	——	——	[54]
	Drop-seq	拟南芥	根	374	——	——	[55]