The application of single-cell sequencing in plant research
Cell is the most basic structural unit of life,and its division,differentiation,and function are determined by gene expression.For a long time,the research on gene expression has been mainly analyzed at the whole tissue or organ level by conventional transcriptome sequencing(RNA-seq),but this method may lead to the equalization of different cells,and neglect the heterogeneity.In recent years,high-resolution single-cell transcriptome sequencing(scRNA-Seq)based on high throughput sequencing technology has been developed,which can isolate and sequence single cells from the same tissue sample in the same experiment.Single-cell transcriptome sequencing can accurately identify cell types and distinguish cell differentiation status according to the gene expression profiles,reveal changes in gene expression at single cell level in tissues and analyze tissue heterogeneity.Therefore scRNA-Seq can provide a new insight into the molecular mechanism of cell differentiation in organisms.The first step of single-cell sequencing is to separate single cells.A series of methods have been developed to separate single cells,including enzymatic hydrolysis,separation of single cell nucleus,laser capture microdissection,dilution,fluorescence-activated cell sorting(FACS),droplet microfluidic control and BD Rhapsody microhole capture,etc.Different separation approaches have advantages and disadvantages,which are analyzed in this review,and can be selected according to different experimental requirements and conditions.Compared with animal cells,the acquisition of a plant single cell needs to be achieved by the preparation of protoplasts.Enzymatic hydrolysis is one of the most commonly used methods to obtain protoplasts,but the process of enzymatic hydrolysis may have an obvious bias to the expression of related genes,and some plant protoplasts with larger diameter may lead to blocking of sequencing equipment,which limits the application of scRNA-Seq in plants.Despite this,the research of single-cell transcriptome in plants has increased rapidly in the past two years,and the combined analysis of multiple omics data has become a trend.This paper reviews the development process of single-cell sequencing,the isolation methods of single-cell,and summarizes the advantages and disadvantages of various methods.Moreover,we introduce the significant progress of single-cell RNA sequencing technology in revealing plant cell development and differentiation processes,explaining biosynthetic pathways,and responding to biotic and abiotic stress in recent years.In addition,scRNA-Seq has played an important role in analyzing the differentiation of cell types between different species.The combined application of multiple omics studies not only helps us to have a deeper understanding of plant biology but also provides new research directions for fields such as plant breeding.We hope to provide a reference for the widespread single-cell level research in more species in the future.
single-cell transcriptomics sequencingsingle cell isolationmultiple omicsplant research
万方数据
维普
