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Updated: Jun 24, 2025

Efficient Agroinfiltration of Plants for High-level Transient Expression of Recombinant Proteins

Efficient Agroinfiltration of Plants for High-level Transient Expression of Recombinant Proteins

Published on: July 23, 2013

Robust soybean leaf agroinfiltration.

Bryce N Trull^1,2, Mst Shamira Sultana¹, Alexander C Pfotenhauer²

¹Department of Plant Sciences, University of Tennessee, Knoxville, TN, USA.

Plant Cell Reports

|June 5, 2024

Summary

This summary is machine-generated.

We developed a robust method for transient gene expression in soybean leaves using agroinfiltration. This technique optimizes factors like plant genotype and leaf age for successful gene delivery in this important crop.

Keywords:

Agroinfiltration Betalain RUBY Reporter gene Soybean Transient gene expression

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Optimization and Utilization of Agrobacterium-mediated Transient Protein Production in Nicotiana

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Published on: April 19, 2014

Area of Science:

Plant Biotechnology
Molecular Biology
Crop Science

Background:

Agrobacterium-mediated agroinfiltration is a key transient assay in dicot species like Nicotiana benthamiana.
Testing gene constructs directly in crop species is desirable for agricultural biotechnology.
Soybean (Glycine max) is a globally significant dicot crop, necessitating efficient gene expression tools.

Purpose of the Study:

To develop a robust agroinfiltration method for transient gene expression in soybean leaves.
To identify critical parameters influencing the success of agroinfiltration in Glycine max.
To provide a new research tool for soybean biotechnology and synthetic biology.

Main Methods:

Optimization of Agrobacterium culture conditions and expression vectors.
Use of Silwet L-77 surfactant and vacuum infiltration (5 min at -85 kPa).
Selection of fully expanded leaves at the VC or V1 growth stage, followed by dark incubation.

Main Results:

Successful high-level expression of GUS, fluorescent proteins, and the RUBY reporter (betalain) in soybean leaves.
Identification of key factors for successful soybean agroinfiltration: genotype, surfactant, developmental stage, and Agrobacterium conditions.
Demonstration of the method's efficacy in two soybean lines (V17-0799DT and TN16-5004).

Conclusions:

A reliable agroinfiltration protocol for soybean leaf transient gene expression has been established.
Plant genotype, leaf age, surfactant, and Agrobacterium parameters are crucial for successful agroinfiltration.
Leaf developmental stage is speculated to be the most critical factor for optimizing agroinfiltration in crop species.