Abstract

Geminiviruses (family Geminiviridae) constitute the largest group of known plant viruses with single-stranded circular DNAs that cause devastating diseases in many economically important crops worldwide. Developing and generating resistant germplasm resources is crucial to prevent and control these devastating diseases. Here, we employed CRISPR/Cas9 system to confer plant resistance against geminiviruses through two strategies. One strategy is to target and edit the viral genome sequences (iterons). The other is to discover and manipulate novel host recessive resistance factors required for the viral infection to block virus invasion. We firstly showed that targeting viral iterons displayed more efficient resistance than targeting viral IRs. However, the specific viral sgRNA could not confer resistance to different geminiviruses. Therefore, targeting viral sequences would not be ideal for engineering resistance against multiple geminiviruses. Then the two novel susceptible genes NbUbEF1B and NbCCR4/NOT3 that might be involved in the replication of the geminiviruses were identified, and knock-out of them through the CRISPR/Cas9 system displayed high-efficiency and broad-spectrum resistance to various geminiviruses. Notably, deficiency of NbUbEF1B did not display any observable growth-defense trade-off. Therefore, editing this recessive resistance gene would have a promising perspective in engineering resistant plants against geminiviruses in the future.

 Plant Biotechnology Journal，IF=13.8

https://pubmed.ncbi.nlm.nih.gov/38147372/