Journal
JOURNAL OF VIROLOGICAL METHODS
Volume 231, Issue -, Pages 38-43Publisher
ELSEVIER
DOI: 10.1016/j.jviromet.2016.02.004
Keywords
Cassava brown streak disease; Cassava brown streak virus; Ugandan cassava brown streak virus; Artificial microRNAs; Virus resistance
Funding
- Uganda-Millennium Science Initiative (MSI) project from the World Bank
- Bill and Melinda Gates Foundation
- United States Agency for International Development from the American people (USAID)
- Monsanto Fund
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Artificial miRNAs (amiRNA) were generated targeting conserved sequences within the genomes of the two causal agents of Cassava brown streak disease (CBSD): Cassava brown streak virus (CBSV) and Ugandan cassava brown streak virus (UCBSV). Transient expression studies on ten amiRNAs targeting 21 nt conserved sequences of P1(CBSV and UCBSV), P3(CBSV and UCBSV), CI(UCBSV), NIb(CBSV and UCBSV), CP(UCBSV) and the un-translated region (3'-UTR) were tested in Nicotiana benthamiana. Four out of the ten amiRNAs expressed the corresponding amiRNA at high levels. Transgenic N. benthamiana plants were developed for the four amiRNAs targeting the P1 and Nib genes of CBSV and the P1 and CP genes of UCBSV and shown to accumulate miRNA products. Transgenic plants challenged with CBSV and UCBSV isolates showed resistance levels that ranged between similar to 20-60% against CBSV and UCBSV and correlated with expression levels of the transgenically derived miRNAs. MicroRNAS targeting P1 and Nib of CBSV showed protection against CBSV and UCBSV, while amiRNAs targeting the P1 and CP of UCBSV showed protection against UCBSV but were less efficient against CBSV. These results indicate a potential application of amiRNAs for engineering resistance to CBSD-causing viruses in cassava. (C) 2016 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
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