The Plant journal : for cell and molecular biologyOct 10, 2024

A cost-effective and efficient genetic editing method for radish using growth regulators and visible markers

CRISPR Gene Editing Weekly Brief ↗PubMed ↗DOI ↗

Updated Mar 22, 2026

Abstract

A highly efficient genetic transformation method for radish has been developed using Agrobacterium rhizogenes-mediated transformation.

The RUBY gene enables a visual red-violet color for effectively monitoring transgenic hairy root screening in radish.
Shoot regeneration capacity is improved by introducing two developmental regulator genes, Wuschel2 (Wus2) and isopentenyl transferase (ipt).
Transgenic shoots can be generated without the addition of external hormones when using the Wus2-ipt combination.
The established transformation system successfully generated gene-edited radish plants by knocking out the phytoene desaturase (RsPDS) gene.
The findings could facilitate genetic transformation and genome editing in other vegetable species.

Simplified

Radish (Raphanus sativus L.) is one of the most important root vegetable crops worldwide. However, gene function exploration and germplasm innovation still face tremendous challenges due to its extremely low transformation efficiency. Here, an economic and highly efficient genetic transformation method for radish was explored by Agrobacterium rhizogenes-mediated transformation with the help of combining special developmental regulator (DR) genes and the visual identification reporter. Firstly, the RUBY gene, a betalain biosynthesis system, could result in a visual red-violet color used as a convenient and effective reporter for monitoring transgenic hairy roots screening of radish. However, the hairy roots-to-shoots conversion system of radish still stands as a barrier to the obtainment of whole transgenic plants, although different hormone combinations and various culture conditions were tried. Following, two DR genes including Wuschel2 (Wus2) and isopentenyl transferase (ipt), as well as their combination Wus2-ipt were introduced for the shoot regeneration capacity improvement. The results showed that the transgenic shoots could be directly generated without externally supplying any hormones in the presence of a Wus2-ipt combination. Then, Wus2-ipt along with the RUBY reporter was employed to establish an efficient genetic transformation system of radish. Moreover, this system was applied in generating gene-edited radish plants and the phytoene desaturase (RsPDS) gene was effectively knockout through albino phenotype observation and sequencing analysis. These findings have the potential to be widely applied in genetic transformation and genome-editing genetic improvement of other vegetable species.

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A cost-effective and efficient genetic editing method for radish using growth regulators and visible markers

Abstract

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