Inducible resistance to maize streak virus

Dionne N. Shepherd, Benjamin Dugdale, Darren P. Martin, Arvind Varsani, Francisco M. Lakay, Marion E. Bezuidenhout, Adérito L. Monjane, Jennifer A. Thomson, James Dale, Edward P. Rybicki

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

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 (pSPLITrepIII-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 pSPLITrepIII-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.

Original languageEnglish (US)
Article numbere105932
JournalPLoS One
Volume9
Issue number8
DOIs
StatePublished - Aug 28 2014
Externally publishedYes

Fingerprint

Maize streak virus
Viruses
Transgenes
transgenes
Zea mays
Geminiviridae
Genes
Mastrevirus
corn
Cell culture
Assays
cell culture
genes
Cell Culture Techniques
genetically modified organisms
circular DNA
Circular DNA
Tissue culture
single-stranded DNA
Proteins

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Shepherd, D. N., Dugdale, B., Martin, D. P., Varsani, A., Lakay, F. M., Bezuidenhout, M. E., ... Rybicki, E. P. (2014). Inducible resistance to maize streak virus. PLoS One, 9(8), [e105932]. https://doi.org/10.1371/journal.pone.0105932

Inducible resistance to maize streak virus. / Shepherd, Dionne N.; Dugdale, Benjamin; Martin, Darren P.; Varsani, Arvind; Lakay, Francisco M.; Bezuidenhout, Marion E.; Monjane, Adérito L.; Thomson, Jennifer A.; Dale, James; Rybicki, Edward P.

In: PLoS One, Vol. 9, No. 8, e105932, 28.08.2014.

Research output: Contribution to journalArticle

Shepherd, DN, Dugdale, B, Martin, DP, Varsani, A, Lakay, FM, Bezuidenhout, ME, Monjane, AL, Thomson, JA, Dale, J & Rybicki, EP 2014, 'Inducible resistance to maize streak virus', PLoS One, vol. 9, no. 8, e105932. https://doi.org/10.1371/journal.pone.0105932
Shepherd DN, Dugdale B, Martin DP, Varsani A, Lakay FM, Bezuidenhout ME et al. Inducible resistance to maize streak virus. PLoS One. 2014 Aug 28;9(8). e105932. https://doi.org/10.1371/journal.pone.0105932
Shepherd, Dionne N. ; Dugdale, Benjamin ; Martin, Darren P. ; Varsani, Arvind ; Lakay, Francisco M. ; Bezuidenhout, Marion E. ; Monjane, Adérito L. ; Thomson, Jennifer A. ; Dale, James ; Rybicki, Edward P. / Inducible resistance to maize streak virus. In: PLoS One. 2014 ; Vol. 9, No. 8.
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