Strategies for the Construction of Cassava Brown Streak Disease Viral Infectious Clones

C. R.A. Duff-Farrier, D. R. Mbanzibwa, S. Nanyiti, Bunawan Hamidun, J. L. Pablo-Rodriguez, K. R. Tomlinson, A. M. James, T. Alicai, S. E. Seal, A. M. Bailey, G. D. Foster

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Cassava brown streak disease (CBSD) has major impacts on yield and quality of the tuberous roots of cassava in Eastern and Central Arica. At least two Potyviridae species cause the disease: Cassava brown streak virus (CBSV) and Ugandan cassava brown streak virus (UCBSV). Cloned viral genome sequences known as infectious clones (ICs) have been important in the study of other viruses, both as a means of standardising infectious material and characterising viral gene function. IC construction is often technically challenging for Potyviridae due to sequence instability in E. coli. Here, we evaluate three methods for the construction of infectious clones for CBSD. Whilst a simple IC for in vitro transcription was made for UCBSV isolate ‘Kikombe’, such an approach failed to deliver full-length clones for CBSV isolates ‘Nampula’ or ‘Tanza’, necessitating more complex approaches for their construction. The ICs successfully generated symptomatic infection in the model host N. benthamiana and in the natural host cassava. This shows that whilst generating ICs for CBSV is still a technical challenge, a structured approach, evaluating both in vitro and in planta transcription systems should successfully deliver ICs, allowing further study into the symptomology and virulence factors in this important disease complex.

Original languageEnglish
JournalMolecular Biotechnology
DOIs
Publication statusAccepted/In press - 1 Jan 2018

Fingerprint

Manihot
Virus Diseases
Viruses
Communicable Diseases
Clone Cells
Potyviridae
Transcription
Genes
Virulence Factors
Escherichia coli
Viral Genes
Viral Genome

Keywords

  • Cassava
  • Cassava brown streak virus
  • Infectious clones
  • Ugandan cassava brown streak virus
  • Virus

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Molecular Biology

Cite this

Duff-Farrier, C. R. A., Mbanzibwa, D. R., Nanyiti, S., Hamidun, B., Pablo-Rodriguez, J. L., Tomlinson, K. R., ... Foster, G. D. (Accepted/In press). Strategies for the Construction of Cassava Brown Streak Disease Viral Infectious Clones. Molecular Biotechnology. https://doi.org/10.1007/s12033-018-0139-7

Strategies for the Construction of Cassava Brown Streak Disease Viral Infectious Clones. / Duff-Farrier, C. R.A.; Mbanzibwa, D. R.; Nanyiti, S.; Hamidun, Bunawan; Pablo-Rodriguez, J. L.; Tomlinson, K. R.; James, A. M.; Alicai, T.; Seal, S. E.; Bailey, A. M.; Foster, G. D.

In: Molecular Biotechnology, 01.01.2018.

Research output: Contribution to journalArticle

Duff-Farrier, CRA, Mbanzibwa, DR, Nanyiti, S, Hamidun, B, Pablo-Rodriguez, JL, Tomlinson, KR, James, AM, Alicai, T, Seal, SE, Bailey, AM & Foster, GD 2018, 'Strategies for the Construction of Cassava Brown Streak Disease Viral Infectious Clones', Molecular Biotechnology. https://doi.org/10.1007/s12033-018-0139-7
Duff-Farrier, C. R.A. ; Mbanzibwa, D. R. ; Nanyiti, S. ; Hamidun, Bunawan ; Pablo-Rodriguez, J. L. ; Tomlinson, K. R. ; James, A. M. ; Alicai, T. ; Seal, S. E. ; Bailey, A. M. ; Foster, G. D. / Strategies for the Construction of Cassava Brown Streak Disease Viral Infectious Clones. In: Molecular Biotechnology. 2018.
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