Establishment of an efficient Agrobacterium rhizogenes-mediated hairy root transformation system for functional analysis in tomato
- 1Guangdong Provincial Key Laboratory for Plant Epigenetics, College of Life Sciences and Oceanography, Shenzhen University
- 2Ganzhou Sub-Center of National Vegetable Quality Standards Center
- 3College of Physics and Optoelectronic Engineering, Shenzhen University
Abstract
Tomato is an important model for functional genomics in Solanaceae, and elucidating its gene functions provides valuable insights into conserved genetic regulatory networks across dicotyledonous plants. Hairy root transformation is a key technology for functional studies in tomato. However, systematic optimization of its parameters remains limited. Here, we established an efficient Agrobacterium rhizogenes -mediated hairy root transformation system for tomato cv. Micro-Tom. Using the RUBY visual reporter gene, we systematically evaluated multiple parameters, including A. rhizogenes strain, bacterial optical density (OD ₆₀₀ ), inoculation duration, acetosyringone (AS) concentration, and the type and concentration of antibiotics for Agrobacterium eradication. ArQual was identified as the most suitable strain. Optimization revealed that an OD ₆₀₀ of 0.7, inoculation duration of 10 min, and AS concentration of 200 μM, these optimized conditions yielded the highest transformation efficiency of 66.01%. To validate the utility of this system, we first demonstrated that overexpression of SlMYB28 promotes anthocyanin accumulation in stable transgenic lines. We then applied the optimized hairy root transformation system to functionally characterize SlMYB28 , achieving 59.33% transformation efficiency and distinct purple pigmentation in transgenic roots. These results support the positive regulatory role of SlMYB28 in anthocyanin biosynthesis and demonstrate the utility of this system for rapid visual validation of stress-responsive genes. This rapid, efficient, and visually tractable transformation platform provides a useful tool for rapid functional screening and preliminary characterization in tomato.
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BibTeX
@article{zhang2026establishment,
title = {{Establishment of an efficient Agrobacterium rhizogenes-mediated hairy root transformation system for functional analysis in tomato}},
author = {Siyi Zhang and Longzhu Huang and Junjie Liu and Zhimin Yin and Ziwei Li and Yuanhui Song and Qiuling Zhang and Tengbo Huang},
journal = {Frontiers in Plant Science},
year = {2026},
month = oct,
doi = {10.3389/fpls.2026.1968539},
url = {https://www.frontiersin.org/articles/10.3389/fpls.2026.1968539/full},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Establishment of an efficient Agrobacterium rhizogenes-mediated hairy root transformation system for functional analysis in tomato
AU - Siyi Zhang
AU - Longzhu Huang
AU - Junjie Liu
AU - Zhimin Yin
AU - Ziwei Li
AU - Yuanhui Song
AU - Qiuling Zhang
AU - Tengbo Huang
PY - 2026
DA - 2026/10/14
JO - Frontiers in Plant Science
DO - 10.3389/fpls.2026.1968539
UR - https://www.frontiersin.org/articles/10.3389/fpls.2026.1968539/full
AB - Tomato is an important model for functional genomics in Solanaceae, and elucidating its gene functions provides valuable insights into conserved genetic regulatory networks across dicotyledonous plants. Hairy root transformation is a key technology for functional studies in tomato. However, systematic optimization of its parameters remains limited. Here, we established an efficient Agrobacterium rhizogenes -mediated hairy root transformation system for tomato cv. Micro-Tom. Using the RUBY visual reporter gene, we systematically evaluated multiple parameters, including A. rhizogenes strain, bacterial optical density (OD ₆₀₀ ), inoculation duration, acetosyringone (AS) concentration, and the type and concentration of antibiotics for Agrobacterium eradication. ArQual was identified as the most suitable strain. Optimization revealed that an OD ₆₀₀ of 0.7, inoculation duration of 10 min, and AS concentration of 200 μM, these optimized conditions yielded the highest transformation efficiency of 66.01%. To validate the utility of this system, we first demonstrated that overexpression of SlMYB28 promotes anthocyanin accumulation in stable transgenic lines. We then applied the optimized hairy root transformation system to functionally characterize SlMYB28 , achieving 59.33% transformation efficiency and distinct purple pigmentation in transgenic roots. These results support the positive regulatory role of SlMYB28 in anthocyanin biosynthesis and demonstrate the utility of this system for rapid visual validation of stress-responsive genes. This rapid, efficient, and visually tractable transformation platform provides a useful tool for rapid functional screening and preliminary characterization in tomato.
SN - 1664-462X
ER -