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. 2026 Feb 26;45(3):66.
doi: 10.1007/s00299-026-03749-1.

Visualizing application of RUBY-MAT system in maize genetic transformation and gene expression pattern detection

Tengfei Jia #  1   2 , Shuai Zhao #  2 , Xiangyu Zhang  1   2 , Yingxue Li  2 , Kuan Cheng  1   2 , Xuehui Qian  2 , Mingli Song  2 , Wenjing Ge  1   2 , Qian Liu  2 , Haoyue Xu  1   2 , Fashuai Li  1   2 , Jinhua Hu  2 , Qi Hao  1   2 , Yanan Tian  3 , Baozhu Li  4   5
Affiliations

Visualizing application of RUBY-MAT system in maize genetic transformation and gene expression pattern detection

Tengfei Jia et al. Plant Cell Rep. .

Abstract

RUBY-MAT system, integrating the RUBY reporter and MAT module, was innovatively applied to Agrobacterium-mediated maize transformation, and precise spatiotemporal gene expression analysis. Maize genetic improvement relies on efficient genetic transformation and gene expression monitoring technologies. In this study, we provided RUBY-MAT system with RUBY reporter and MAT (morphogene-assisted transformation), and realized its visual application in Agrobacterium-mediated genetic transformation of maize immature embryos and gene expression patterns. Experimental results demonstrated that RUBY-MAT system can be successfully applied in Agrobacterium-mediated genetic transformation of maize immature embryos, and RUBY reporter enables non-invasive monitoring of positive transformation events, as early as 7 days after culture, through the clearly distinguishable betalain of positive callus. Dynamic expression monitoring further revealed that RUBY reporter can reflect the spatiotemporal expression patterns of specific promoters in real time: in constitutively expressed RUBY plants, high activity was observed in tissues including tassels, anthers, and stem nodes; in ZmH1A promoter-driven RUBY lines, betalain accumulation clearly indicated gene expression in vascular bundles and stomatal guard cells; while in lines harboring the putative young leaf-specific ZmGLP1 promoter, betalain specifically accumulated in young leaves. Quantitative analysis confirmed a strong linear correlation between betalain absorbance and promoter activity, establishing the RUBY reporter as a reliable quantitative tool for monitoring transformation efficiency and gene expression at tissue and cellular levels. Collectively, the RUBY-MAT system offers robust technical support for maize genetic transformation and functional gene analysis, demonstrating broad application potential in both field and laboratory studies.

Keywords: Betalain; Genetic transformation; Maize; RUBY-MAT.

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Conflict of interest statement

Declarations. Conflict of interest: Authors declare that there are no competing interests associated with this work.

References

    1. Aesaert S, Impens L, Coussens G, Van Lerberge E, Vanderhaeghen R, Desmet L, Vanhevel Y, Bossuyt S, Wambua AN, Van Lijsebettens M, Inzé D, De Keyser E, Jacobs TB, Karimi M, Pauwels L (2022) Optimized transformation and gene editing of the B104 public maize inbred by improved tissue culture and use of morphogenic regulators. Front Plant Sci 13:883847. https://doi.org/10.3389/fpls.2022.883847 - DOI - PubMed - PMC
    1. Alok A, Raman V, D’Agostino L, Kshetry AO, Rai KM, Wang C, Gunapati S, Stupar RM, Patil GB, Zhang F (2025) Developmental regulators enable rapid and efficient soybean transformation and CRISPR-mediated genome editing. Plant Physiol 10:kiaf640. https://doi.org/10.1093/plphys/kiaf640 - DOI
    1. Anand A, Bass SH, Wu E, Wang N, McBride KE, Annaluru N, Miller M, Hua M, Jones TJ (2018) An improved ternary vector system for Agrobacterium-mediated rapid maize transformation. Plant Mol Biol 97:187–200. https://doi.org/10.1007/s11103-018-0732-y - DOI - PubMed - PMC
    1. Anjanappa RB, Gruissem W (2021) Current progress and challenges in crop genetic transformation. J Plant Physiol 261:153411. https://doi.org/10.1016/j.jplph.2021.153411 - DOI - PubMed
    1. Bonifácio-Anacleto F, Martin JABS, Reutemann AG, Habermann E, Pozner RE, Nazareno AG, Nogueira FM, Martinez CA, Alzate-Marin AL (2024) Warming and water deficit impact the reproductive features of the tropical forage species Stylosanthes capitata. Environ Exp Bot 226(830):105899. https://doi.org/10.1016/j.envexpbot.2024.105899 - DOI

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