Development of a rice yellow mottle virus amplicon biotechnological tool for the production of high value proteins in rice

Lately, advances on biotechnology have led to the emergence of plants as bioreactor for the production of molecules of industrial, pharmaceutical and agribusiness interests. One of the major advances was the exploitation of plant viruses as “amplifier” for the protein production. This amplicon technology relies on viral vector engineered to contain a gene encoding candidate protein that is produced in significant quantities during virus replication. To date, only few of these virus tools are developed for monocot species. Here, we set up experimental approaches to develop the first amplicon tool based on the Rice yellow mottle virus (RYMV) to be used on rice. We showed that inoculated 4-week-old leaves display the most important RYMV initial replication and retained this tissue for protein of interest production from RYMV amplicon. We engineered RYMV tool based on highly replicative RYMVMg1 isolated with GFP gene of interest replacing Coat Protein ORF. Replication of RYMV amplicon was optimized in inoculated rice leaves by adding an RYMV helper to complement RYMV amplicon with CP. This work identify for the first time a viral amplicon tool active in rice offering promising perspectives for the use of rice as bioreactor for the production of high value proteins.   Key words: Rice, rice yellow mottle virus, plant bioreactor, viral amplicon, virus induced gene silencing (VIGS).