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

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

*Corresponding author for this work

Research output: Contribution to journalArticle (Academic Journal)

5 Citations (Scopus)
215 Downloads (Pure)

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
Pages (from-to)93-101
Number of pages9
JournalMolecular Biotechnology
Volume61
Issue number2
Early online date27 Nov 2018
DOIs
Publication statusPublished - 15 Feb 2019

Keywords

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

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    Duff-Farrier, C. R. A., Mbanzibwa, D. R., Nanyiti, S., Bunawan, H., Pablo-Rodriguez, J. L., Tomlinson, K. R., James, A. M., Alicai, T., Seal, S. E., Bailey, A. M., & Foster, G. D. (2019). Strategies for the Construction of Cassava Brown Streak Disease Viral Infectious Clones. Molecular Biotechnology, 61(2), 93-101. https://doi.org/10.1007/s12033-018-0139-7