ArticlePlant methods2012
FACS-based purification of Arabidopsis microspores, sperm cells and vegetative nuclei.
Article in Plant methods, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 51 papers.
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Who cites it
51 citing papers in PubMed, 93 citations in OpenAlex.
- Adaptive response to long-term high temperatures during the reproductive development in Arabidopsis thaliana.Planta · 2026Article
- ClPS1 gene-mediated manipulation of 2n pollen formation enables the creation of triploid seedless watermelon.Molecular horticulture · 2025Article
- Kinesins control male germ unit assembly for sperm delivery in Arabidopsis.Nature plants · 2025Article
- Dynamic changes in 3D chromatin structure during male gametogenesis in Arabidopsis thaliana.Genome biology · 2025Article
- Pseudouridine guides germline small RNA transport and epigenetic inheritance.Nature structural & molecular biology · 2025Article
- Quantitative and spatially resolved detection of multiplexed microRNA from plant tissue via hybridization to hydrogel-bound DNA probes in nanoliter well arrays.Microsystems & nanoengineering · 2024Article
- Assessing data analysis techniques in a high-throughput meiosis-like induction detection system.Plant methods · 2024Article
- A cell-based fluorescent system and statistical framework to detect meiosis-like induction in plants.Frontiers in plant science · 2024Article
- Cell cycle status of male and female gametes during Arabidopsis reproduction.Plant physiology · 2023Article
- Cell-type-specific alternative splicing in the Arabidopsis germline.Plant physiology · 2023Article
- Overcoming roadblocks forFrontiers in plant science · 2023Review
- RNA-seq analysis of synchronized developing pollen isolated from a single anther.Frontiers in plant science · 2023Article
- Evolutionary Relationships and Divergence ofGenes · 2022Article
- DNA methylation dynamics during germline development.Journal of integrative plant biology · 2022Review
- Single-nucleus RNA-seq reveals that MBD5, MBD6, and SILENZIO maintain silencing in the vegetative cell of developing pollen.Cell reports · 2022Article
- Article
- H3K27 methylation regulates the fate of two cell lineages in male gametophytes.The Plant cell · 2022Article
- The poly(A) polymerase PAPS1 mediates pollen maturation by regulating sperm cell differentiation in plants.Plant direct · 2022Article
- Cellular export of sugars and amino acids: role in feeding other cells and organisms.Plant physiology · 2021Review
- Defects in meiotic chromosome segregation lead to unreduced male gametes in Arabidopsis SMC5/6 complex mutants.The Plant cell · 2021Article
Corrections and comments
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Authors and funding
8 authors at 3 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
unlabelled
backgroundThe male germline in flowering plants differentiates by asymmetric division of haploid uninucleated microspores, giving rise to a vegetative cell enclosing a smaller generative cell, which eventually undergoes a second mitosis to originate two sperm cells. The vegetative cell and the sperm cells activate distinct genetic and epigenetic mechanisms to control pollen tube growth and germ cell specification, respectively. Therefore, a comprehensive characterization of these processes relies on efficient methods to isolate each of the different cell types throughout male gametogenesis.
resultsWe developed stable transgenic Arabidopsis lines and reliable purification tools based on Fluorescence-Activated Cell Sorting (FACS) in order to isolate highly pure and viable fractions of each cell/nuclei type before and after pollen mitosis. In the case of mature pollen, this was accomplished by expressing GFP and RFP in the sperm and vegetative nuclei, respectively, resulting in 99% pure sorted populations. Microspores were also purified by FACS taking advantage of their characteristic small size and autofluorescent properties, and were confirmed to be 98% pure.
conclusionsWe provide simple and efficient FACS-based purification protocols for Arabidopsis microspores, vegetative nuclei and sperm cells. This paves the way for subsequent molecular analysis such as transcriptomics, DNA methylation analysis and chromatin immunoprecipitation, in the developmental context of microgametogenesis in Arabidopsis.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.