An Efficient Cell Sorting Protocol for Maize Protoplasts
Carlos Ortiz‐Ramíreza b, Edgar Demesa-Arévaloc d, Xiaosa Xuc d, David P. Jacksonc d, Kenneth D. Birnbauma b✉
- aCenter for Genomics and Systems Biology, Department of Biology, New York University New York New York
- bCenter for Genomics and Systems Biology, Department of Biology, New York University, New York, New York
- cCold Spring Harbor Laboratory, Cold Spring Harbor New York
- dCold Spring Harbor Laboratory, Cold Spring Harbor, New York
- ✉Corresponding author
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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. © 2018 by John Wiley & Sons, Inc.
TL;DRA new protocol is reported that dramatically increases protoplast yield from maize for subsequent cell isolation by FACS, efficient in generating protoplast from root and shoot inner layers and can be applied successfully to Arabidopsis thaliana. (Semantic Scholar)