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Related Experiment Video

Updated: Jun 17, 2026

A Robotic Platform for High-throughput Protoplast Isolation and Transformation

A Robotic Platform for High-throughput Protoplast Isolation and Transformation

Published on: September 27, 2016

Systematic Optimization Enables Near-Perfect In Vitro Transformation Efficiencies for Spirodela polyrhiza (Greater

Tasmia Islam¹, Ayalew Ligaba-Osena², Eric A Josephs^1,2,3

¹Department of Nanoscience; The Joint School of Nanoscience and Nanoengineering (JSNN); UNC Greensboro; Greensboro, NC 27402.

Biorxiv : the Preprint Server for Biology

|September 26, 2025

Summary

We optimized plant transformation protocols for Spirodela polyrhiza (Greater Duckweed), achieving high efficiency in weeks. This breakthrough enables rapid genetic modification for agricultural and biotechnological applications.

Keywords:

CRISPR/Cas9 Duckweed Gene editing Spirodela polyrhiza Stable transformation Tissue culture Transient transformation morphogenic regulators

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Last Updated: Jun 17, 2026

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Area of Science:

Plant Biotechnology
Molecular Biology
Genomics

Background:

In vitro plant transformation is crucial for crop improvement and understanding gene function.
Existing transformation methods are often inefficient and time-consuming for many plant species.
Duckweeds (Spirodela polyrhiza) show promise for biotechnology but have challenging transformation efficiencies.

Purpose of the Study:

To develop and optimize a streamlined, high-efficiency in vitro transformation protocol for Spirodela polyrhiza.
To overcome bottlenecks in callus induction, genetic transfection, and plant regeneration.
To enable rapid functional genomics and synthetic biology applications in duckweed.

Main Methods:

Integrated optimization of callus induction, transient and stable genetic transformation, and regeneration.
Development of visual marker-free selection for transformants.
Validation of stable transgene expression over 100 generations.

Main Results:

Achieved >90-100% efficiency for callus induction, transformation, and regeneration.
Reduced transformation timeline from months to weeks for Spirodela polyrhiza.
Demonstrated stable transgene expression for over 100 generations in regenerated plants.

Conclusions:

The optimized protocol significantly enhances Spirodela polyrhiza transformation efficiency and speed.
This streamlined approach facilitates high-throughput functional genomics and synthetic biology.
The method provides a robust platform for biotechnological applications using duckweed.