Development of a Mini-Replicon-Based Reverse-Genetics System for Rice Stripe Tenuivirus

Area of Science:

• Plant virology
• Molecular biology
• RNA virus research

Background:

• Negative-stranded RNA (NSR) viruses cause significant diseases in plants and animals.
• Rice stripe tenuivirus (RSV) is a major destructive pathogen of rice in Asia.
• Lack of reverse-genetics systems hinders molecular studies of RSV.

Purpose of the Study:

• To develop a mini-replicon-based reverse-genetics system for RSV.
• To enable functional analysis of RSV genes and their interactions with host plants.
• To provide a tool for studying RSV replication, transcription, and movement.

Main Methods:

• Developed a mini-replicon system expressing an RSV genomic RNA3 reporter and essential proteins (NP, RdRpopt).
• Utilized Nicotiana benthamiana as a host for system development and validation.
• Constructed mini-replicons for all four RSV genomic RNAs.

Main Results:

• RSV nucleocapsid (NP) and RNA-dependent RNA polymerase (RdRpopt) are essential for viral RNA expression.
• Viral suppressors of RNA silencing (VSRs) enhanced reporter gene expression.
• The movement protein (NSvc4) facilitated cell-to-cell trafficking of viral RNA.

Conclusions:

• This study presents the first mini-replicon-based reverse-genetics system for tenuivirus.
• The developed system is crucial for elucidating molecular mechanisms of tenuivirus infection in plants.
• This platform will advance research on RSV and other segmented NSR viruses affecting crops.