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<p>Plants engineered with synthetic genetic programs can transform how we monitor and manage the extension of crop pests and diseases. Here, we establish a bioluminescent platform in <i>Nicotiana benthamiana</i> for autonomous viral sensing based on the fungal bioluminescence pathway (FBP). We first demonstrate that recombinant viruses can deliver missing pathway components, establishing a methodological framework for spatially resolved tracking of infection dynamics. Leveraging this starting point, we develop a dual-output sentinel circuit that uses a protease-responsive bioluminescence resonance energy transfer (BRET) module to report infection through a virus-triggered spectral shift in luminescence. In the absence of infection, plants emit a stable yellow glow indicating system integrity. Upon infection with potyviruses, cleavage of the BRET fusion by the virus-encoded NIa-protease activates a distinct colour change detectable with low-cost imaging. This modular design is compatible with other pathogens carrying specific proteases and supports future multiplexing strategies. Our results highlight the potential of synthetic sentinel gene circuits as autonomous biosensors for precision crop protection.</p>

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Bioluminescent sentinel plants enable autonomous diagnostics of viral infections

  • 期刊论文

  • 出版模式：

    开放获取

  • 发表日期：

    2026年5月29日

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  • Camilo Calvache,
  • Marta Rodriguez-Rodriguez,
  • Victor Vazquez-Vilriales,
  • Elena Garcia-Perez,
  • Aubin Fleiss,
  • Mustafá Ezzeddin-Ayoub,
  • Fabio Pasin,
  • José-Antonio Daròs,
  • Karen S. Sarkisyan,
  • Diego Orzaez,
  • Marta Vazquez-Vilar

摘要

Plants engineered with synthetic genetic programs can transform how we monitor and manage the extension of crop pests and diseases. Here, we establish a bioluminescent platform in Nicotiana benthamiana for autonomous viral sensing based on the fungal bioluminescence pathway (FBP). We first demonstrate that recombinant viruses can deliver missing pathway components, establishing a methodological framework for spatially resolved tracking of infection dynamics. Leveraging this starting point, we develop a dual-output sentinel circuit that uses a protease-responsive bioluminescence resonance energy transfer (BRET) module to report infection through a virus-triggered spectral shift in luminescence. In the absence of infection, plants emit a stable yellow glow indicating system integrity. Upon infection with potyviruses, cleavage of the BRET fusion by the virus-encoded NIa-protease activates a distinct colour change detectable with low-cost imaging. This modular design is compatible with other pathogens carrying specific proteases and supports future multiplexing strategies. Our results highlight the potential of synthetic sentinel gene circuits as autonomous biosensors for precision crop protection.