An Efficient Cell Sorting Protocol for Maize Protoplasts

Carlos Ortiz-Ramírez, Edgar Demesa Arevalo, Xiaosa Xu, David P. Jackson, Kenneth D. Birnbaum

Research output: Contribution to journalArticlepeer-review

Abstract

Maize is one the most widely cultivated crops worldwide and an important model system for the study of genetics and cytogenetics. Although the availability of a genome sequence has enabled new quantitative genomic studies, developing methods to isolate specific types of cells will enable useful approaches for transcriptomic analysis in the crop plant. Fluorescence-activated cell sorting (FACS) is a powerful technique for cell isolation and the study of transcriptional profiles from specific cell populations. The use of FACS on plant cells requires the generation of protoplasts by tissue digestion and cell wall removal. Although some protocols are available, they mainly focus on dicot species and obtaining sufficient protoplasts from inner tissue layers has been challenging in both monocots and dicots. Here, we report a new protocol that dramatically increases protoplast yield from maize for subsequent cell isolation by FACS. This protocol is efficient in generating protoplasts from root and shoot inner layers and can also be applied successfully to Arabidopsis thaliana.

Original languageEnglish (US)
Pages (from-to)e20072
JournalCurrent protocols in plant biology
Volume3
Issue number3
DOIs
StatePublished - Sep 1 2018

Keywords

  • FACS
  • maize
  • monocots
  • protoplasts
  • transcriptome

ASJC Scopus subject areas

  • Medicine(all)

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