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Key message <p>RUBY-MAT system, integrating the <i>RUBY</i> reporter and MAT module, was innovatively applied to Agrobacterium-mediated maize transformation, and precise spatiotemporal gene expression analysis.</p> Abstract <p>Maize genetic improvement relies on efficient genetic transformation and gene expression monitoring technologies. In this study, we provided RUBY-MAT system with <i>RUBY</i> 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 <i>RUBY</i> reporter enables non-invasive monitoring of positive transformation events, as early as 7&#xa0;days after culture, through the clearly distinguishable betalain of positive callus. Dynamic expression monitoring further revealed that <i>RUBY</i> reporter can reflect the spatiotemporal expression patterns of specific promoters in real time: in constitutively expressed <i>RUBY</i> plants, high activity was observed in tissues including tassels, anthers, and stem nodes; in <i>ZmH1A</i> promoter-driven <i>RUBY</i> lines, betalain accumulation clearly indicated gene expression in vascular bundles and stomatal guard cells; while in lines harboring the putative young leaf-specific <i>ZmGLP1</i> 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.</p>

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

  • 期刊论文

  • 发表日期：

    2026年2月26日

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  • Tengfei Jia,
  • Shuai Zhao,
  • Xiangyu Zhang,
  • Yingxue Li,
  • Kuan Cheng,
  • Xuehui Qian,
  • Mingli Song,
  • Wenjing Ge,
  • Qian Liu,
  • Haoyue Xu,
  • Fashuai Li,
  • Jinhua Hu,
  • Qi Hao,
  • Yanan Tian,
  • Baozhu Li

摘要

Key message

RUBY-MAT system, integrating the RUBY reporter and MAT module, was innovatively applied to Agrobacterium-mediated maize transformation, and precise spatiotemporal gene expression analysis.

Abstract

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.