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Inhibition of maize streak virus (MSV) replication by transient and transgenic expression of MSV replication-associated protein mutants

Jennifer A. Thomson^1,

Edward P. Rybicki^1,3,

¹ Department of Molecular and Cell Biology, University of Cape Town, Private Bag, Rondebosch, Cape Town 7701, South Africa
² Division of Pharmacology, University of Cape Town, Cape Town, South Africa
³ Institute of Infectious Disease and Molecular Medicine, University of Cape Town, Observatory, Anzio Road, Cape Town 7925, South Africa

Correspondence
Edward P. Rybicki
ed{at}science.uct.ac.za

Maize streak disease is a severe agricultural problem in Africa and the development of maize genotypes resistant to the causal agent, Maize streak virus (MSV), is a priority. A transgenic approach to engineering MSV-resistant maize was developed and tested in this study. A pathogen-derived resistance strategy was adopted by using targeted deletions and nucleotide-substitution mutants of the multifunctional MSV replication-associated protein gene (rep). Various rep gene constructs were tested for their efficacy in limiting replication of wild-type MSV by co-bombardment of maize suspension cells together with an infectious genomic clone of MSV and assaying replicative forms of DNA by quantitative PCR. Digitaria sanguinalis, an MSV-sensitive grass species used as a model monocot, was then transformed with constructs that had inhibited virus replication in the transient-expression system. Challenge experiments using leafhopper-transmitted MSV indicated significant MSV resistance – from highly resistant to immune – in regenerated transgenic D. sanguinalis lines. Whereas regenerated lines containing a mutated full-length rep gene displayed developmental and growth defects, those containing a truncated rep gene both were fertile and displayed no growth defects, making the truncated gene a suitable candidate for the development of transgenic MSV-resistant maize.

These authors contributed equally to this work.