Rapid spray-mediated plant expression platform for functional screening of herbicide-tolerant protein variants
A rapid plant-based screening platform that transiently expresses engineered protein variants in Nicotiana benthamiana via simple spray delivery, enabling functional testing of herbicide tolerance in 4-6 days. Accelerates discovery of herbicide-tolerant proteins by bridging high-throughput design and crop validation, prioritizing promising candidates before stable transformation.
Overview
A rapid, scalable plant-based platform for functional screening of engineered protein variants for herbicide tolerance. It builds on an in-house spray-mediated plant expression technology that enables recombinant proteins to be transiently expressed in Nicotiana benthamiana plants within 4-6 days of spray delivery at very high levels. The platform tests libraries of herbicide-target or detoxification protein variants directly in plants, identifying promising candidates before stable transformation and validation in target crops.
By combining natural N. benthamiana ecotype diversity with optimized expression vectors, the system provides a simple, robust, and indefinitely scalable workflow bridging high-throughput protein design and crop validation, reducing timelines and costs compared to conventional stable transformation-based screening.
Technical specifications
How it works:
- Candidate genes are cloned into selected expression vectors, with targeting signal peptides added when required.
- Vectors are transferred into Agrobacterium and delivered to a highly spray-efficient N. benthamiana ecotype via spray application.
- Multiple protein variants are expressed in parallel and assessed for expression, folding, enzymatic activity, and reproducibility.
- Sprayed plants are challenged in vivo with selected herbicides and evaluated for visual phenotypes, photosynthetic damage, and metabolite profiles.
Key features:
- Protein expression in 4-6 days after spray delivery.
- High-level production of complex recombinant proteins at low cost.
- Indefinitely scalable spray delivery, unlike vacuum infiltration methods.
- Uses a specially characterized N. benthamiana ecotype from over a decade of wild ecotype research.
Technology readiness level
The underlying spray-mediated expression technology is established in-house, including identification of a highly spray-efficient N. benthamiana ecotype. The proposed work adapts this platform into a herbicide-tolerance screening workflow through two work packages: high-throughput protein expression (months 1-4) and in planta herbicide challenge (months 5-10). The platform is at an early-to-mid development stage for this application, with validation planned using wild-type plants and known variants before candidate screening deployment.
About Queensland University of Technology
Queensland University of Technology (QUT) is a major Australian public research university in Brisbane with two inner-city campuses and a practical, industry-focused culture. Industry partners tap shared core laboratories and technical staff through the Central Analytical Research Facility, and collaborate with QUT groups at the Translational Research Institute alongside hospital partners. A dedicated Office of Industry Engagement brokers collaborative and contract research, while entrepreneurship programs and commercialization pathways help convert university IP into ventures and licenses. Research is sustained by competitive national funding, including the Australian Research Council and National Health and Medical Research Council, with added support from programs such as the Medical Research Future Fund.
delivered to your inbox.