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Related Experiment Video

Updated: May 5, 2026

Scalable Transfection of Maize Mesophyll Protoplasts

Scalable Transfection of Maize Mesophyll Protoplasts

Published on: June 23, 2023

High frequency callus formation from maize protoplasts.

S R Ludwig¹, D A Somers, W L Petersen

¹Department of Biochemistry, University of Minnesota, 55108, St. Paul, MN, USA.

TAG. Theoretical and Applied Genetics. Theoretische Und Angewandte Genetik

|November 20, 2013

Summary

This summary is machine-generated.

A novel solid feeder layer technique significantly enhances maize protoplast callus formation. This method offers a 100- to 1,000-fold improvement over traditional plating, crucial for genetic research.

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Area of Science:

Plant Biotechnology
Molecular Biology
Agricultural Science

Background:

Plant protoplast culture is vital for genetic transformation and mutant recovery in maize.
Conventional methods for maize protoplast culture often yield low callus formation frequencies.
Improving protoplast-derived callus formation is essential for advancing maize breeding and research.

Purpose of the Study:

To develop and evaluate a solid feeder layer technique for enhanced callus formation in Black Mexican Sweet maize (Zea mays L.) protoplast cultures.
To compare the efficiency of the feeder layer method with conventional protoplast plating techniques.
To identify factors influencing callus formation frequency in maize protoplast cultures.

Main Methods:

Protoplasts were plated on a cellulose nitrate filter over agarose-solidified media containing Black Mexican Sweet suspension feeder cells.
Callus colony formation was assessed at various protoplast densities (10^3-10^5 protoplasts/0.2 ml).
The feeder layer method was compared against liquid culture and agarose embedding techniques.

Main Results:

The solid feeder layer technique achieved callus colony formation frequencies exceeding 10%.
This represents a 100- to 1,000-fold increase compared to conventional plating methods.
Higher frequencies were observed across three independently maintained Black Mexican Sweet suspension lines.

Conclusions:

The solid feeder layer technique is a highly effective method for improving maize protoplast callus formation.
This advancement is critical for the successful recovery of rare mutants and genetically transformed maize protoplasts.
Suspension culture maintenance and auxin concentrations can influence callus formation efficiency, warranting further investigation.