Expanding Possibilities for Foreign Gene Expression by Cucumber Green Mottle Mosaic Virus Genome-Based Bipartite Vector System

• Published in: Plants (Basel) Vol. 13; no. 10; p. 1414
• Main Authors: Chattopadhyay, Anirudha, Jailani, A. Abdul Kader, Roy, Anirban, Mukherjee, Sunil Kumar, Mandal, Bikash
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 01.05.2024
• Subjects: Cargo capacity; CGMMV; Cloning; Comparative analysis; Cucumber green mottle mosaic virus; Cucumbers; Cucurbitaceae; deconstructed genomes; Diseases and pests; enzymes; expression; face; Gene expression; gene silencing; Genes; Genetic aspects; genome; Genomes; Genomics; Infiltration; movement; Nicotiana benthamiana; Plant viruses; Plasmids; Proteins; Replicase; Replication; Thermal cycling; trans-replication; Virus diseases of plants; virus-based vector; Viruses; United States; India; deconstructed genomes; CGMMV; expression; trans-replication; movement; virus-based vector; N. benthamiana
• ISSN: 2223-7747; 2223-7747
• DOI: 10.3390/plants13101414

Expanding possibilities for foreign gene expression in cucurbits, we present a novel approach utilising a bipartite vector system based on the cucumber green mottle mosaic virus (CGMMV) genome. Traditional full-length CGMMV vectors face limitations such as a restricted cargo capacity and unstable foreign gene expression. To address these challenges, we developed two ‘deconstructed’ CGMMV genomes, DG-1 and DG-2. DG-1 features a major internal deletion, resulting in the loss of crucial replicase enzyme domains, rendering it incapable of self-replication. However, a staggered infiltration of DG-1 in CGMMV-infected plants enabled successful replication and movement, facilitating gene-silencing experiments. Conversely, DG-2 was engineered to enhance replication rates and provide multiple cloning sites. Although it exhibited higher replication rates, DG-2 remained localised within infiltrated tissue, displaying trans-replication and restricted movement. Notably, DG-2 demonstrated utility in expressing GFP, with a peak expression observed between 6 and 10 days post-infiltration. Overall, our bipartite system represents a significant advancement in functional genomics, offering a robust tool for foreign gene expression in Nicotiana benthamiana.