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Related Experiment Video

Updated: Jan 16, 2026

Rapid, Seamless Generation of Recombinant Poxviruses using Host Range and Visual Selection

Rapid, Seamless Generation of Recombinant Poxviruses using Host Range and Visual Selection

Published on: May 24, 2020

Engineered geminivirus replicons enable rapid in planta directed evolution.

Haocheng Zhu¹, Xu Qin², Leyan Wei^1,3

¹New Cornerstone Science Laboratory, Laboratory of Advanced Breeding Technologies, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, China.

Science (New York, N.Y.)

|October 2, 2025

Summary

Directed evolution in plant cells is now efficient with geminivirus replicon-assisted in planta directed evolution (GRAPE). This new platform rapidly screens genetic variants for enhanced properties, accelerating plant science research.

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Area of Science:

Plant biotechnology
Molecular biology
Genetics

Background:

Directed evolution accelerates the generation of genetic variants with novel properties.
Existing methods for in planta directed evolution are inefficient, limiting high-throughput screening.
Developing efficient platforms for plant cellular evolution is crucial for advancing plant science.

Purpose of the Study:

To develop a novel, high-throughput platform for directed evolution directly within plant cells.
To enable the rapid screening of large libraries of genetic variants for desired functions.
To demonstrate the utility of the platform in evolving plant immune receptors.

Main Methods:

Developed geminivirus replicon-assisted in planta directed evolution (GRAPE).
Linked gene function to geminivirus rolling circle replication for high-throughput selection.
Screened up to 10⁵ variants on *Nicotiana benthamiana* leaves within 4 days.

Main Results:

Successfully evolved the immune receptor NRC3 to resist nematode effector SPRYSEC15 inhibition.
Broadened the recognition spectrum of rice immune receptor Pikm-1 to all six alleles of AVR-Pik.
Demonstrated GRAPE's capability for rapid, scalable, and generalizable evolution in planta.

Conclusions:

GRAPE provides a breakthrough platform for efficient directed evolution in plant cells.
The system significantly accelerates the discovery and engineering of plant genes.
GRAPE is a versatile tool applicable to diverse gene evolution in plants.