Engineered Biomolecular Condensates Limit Tobacco Mosaic Virus Accumulation and Symptom Development

Area of Science:

• Synthetic biology
• Plant virology
• Biomolecular condensates

Background:

• Liquid-liquid phase separation (LLPS) forms artificial condensates studied for cellular function control.
• Cellular condensates can limit viral accumulation, but LLPS for antiviral strategies is underexplored.

Purpose of the Study:

• To engineer artificial condensates for targeted antiviral strategies against tobacco mosaic virus (TMV).
• To assess the efficacy of these condensates in inhibiting TMV accumulation and disease severity in plants.

Main Methods:

• Engineered TMV with MS2 bacteriophage hairpins for specific binding by MS2 coat protein (MCP).
• Developed artificial condensates using intrinsically disordered regions (IDRs) as scaffolds to target MS2-tagged TMV.
• Expressed engineered condensates in Nicotiana benthamiana for transient expression and manual inoculation studies.

Main Results:

• Engineered condensates inhibited MS2-tagged TMV accumulation up to fivefold.
• TDP-43-based condensates reduced MS2-tagged virus accumulation over twofold, independent of translational repression.
• Systemic TMV accumulation reduced over 10-fold in plants expressing TDP-43:MCP condensates, with decreased symptom severity.

Conclusions:

• Artificial biomolecular condensates can effectively target and inhibit plant virus accumulation.
• This approach provides a foundation for developing novel strategies to mitigate plant virus diseases.
• LLPS-based systems offer a tunable and reversible platform for plant disease management.