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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
Inducible resistance to maize streak virus
PLoS ONE, Volume 9, No. 8, Article e105932, Year 2014
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Description
Maize streak virus (MSV), which causes maize streak disease (MSD), is the major viral pathogenic constraint on maize production in Africa. Type member of the Mastrevirus genus in the family Geminiviridae, MSV has a 2.7 kb, single-stranded circular DNA genome encoding a coat protein, movement protein, and the two replication-associated proteins Rep and RepA. While we have previously developed MSV-resistant transgenic maize lines constitutively expressing "dominant negative mutant" versions of the MSV Rep, the only transgenes we could use were those that caused no developmental defects during the regeneration of plants in tissue culture. A better transgene expression system would be an inducible one, where resistance-conferring transgenes are expressed only in MSV-infected cells. However, most known inducible transgene expression systems are hampered by background or "leaky" expression in the absence of the inducer. Here we describe an adaptation of the recently developed INPACT system to express MSV-derived resistance genes in cell culture. Split gene cassette constructs (SGCs) were developed containing three different transgenes in combination with three different promoter sequences. In each SGC, the transgene was split such that it would be translatable only in the presence of an infecting MSV's replication associated protein. We used a quantitative real-time PCR assay to show that one of these SGCs (pSPLITrep
III-Rb-
Ubi) inducibly inhibits MSV replication as efficiently as does a constitutively expressed transgene that has previously proven effective in protecting transgenic maize from MSV. In addition, in our cell-culture based assay pSPLITrep
III-Rb-
Ubi inhibited replication of diverse MSV strains, and even, albeit to a lesser extent, of a different mastrevirus species. The application of this new technology to MSV resistance in maize could allow a better, more acceptable product. © 2014 Shepherd et al.
Available Materials
https://efashare.b-cdn.net/share/pmc/articles/PMC4148390/bin/pone.0105932.s001.doc
Authors & Co-Authors
Shepherd, Dionne Natalie
South Africa, Cape Town
University of Cape Town
Dugdale, Benjamin
Australia, Brisbane
Queensland University of Technology
Martin, Darren Patrick
South Africa, Cape Town
University of Cape Town
South Africa, Cape Town
Centre for High Performance Computing, Cape Town
Varsani, Arvind
New Zealand, Christchurch
University of Canterbury
United States, Gainesville
University of Florida
South Africa, Cape Town
University of Cape Town
Lakay, Francisco M.
South Africa, Cape Town
University of Cape Town
Bezuidenhout, Marion E.
South Africa, Cape Town
University of Cape Town
Monjane, Adérito Luis
South Africa, Cape Town
University of Cape Town
Thomson, Jennifer Ann
South Africa, Cape Town
University of Cape Town
Dale, James L.
Australia, Brisbane
Queensland University of Technology
Rybicki, Edward P.
South Africa, Cape Town
University of Cape Town
Statistics
Citations: 11
Authors: 10
Affiliations: 5
Identifiers
Doi:
10.1371/journal.pone.0105932
e-ISSN:
19326203
Research Areas
Genetics And Genomics
Study Approach
Quantitative