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. ORCID: https://orcid.org/0000-0002-3952-1562, Bailey, A. M. and Foster, G. D. (2018) Strategies for the construction of cassava brown streak disease viral infectious clones. Molecular Biotechnology, 61 (2). pp. 93-101. ISSN 1073-6085 (Print), 1559-0305 (Online) (doi:10.1007/s12033-018-0139-7)

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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.

Item Type:	Article
Additional Information:	© The Author(s) 2018. Open Access. This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.
Uncontrolled Keywords:	Virus, Cassava, Cassava brown streak virus, Ugandan cassava brown streak virus, Infectious clones
Subjects:	S Agriculture > S Agriculture (General)
Faculty / School / Research Centre / Research Group:	Faculty of Engineering & Science Faculty of Engineering & Science > Natural Resources Institute Faculty of Engineering & Science > Natural Resources Institute > Agriculture, Health & Environment Department
Last Modified:	20 Feb 2019 15:38
URI:	http://gala.gre.ac.uk/id/eprint/22280

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