Artificial microRNA-derived resistance to Cassava brown streak disease
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Date
2016
Journal Title
Journal ISSN
Volume Title
Publisher
Journal of virological methods
Abstract
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
∼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.
Description
Keywords
Cassava brown streak disease, Cassava brown streak virus, Ugandan cassava brown streak virus, Artificial microRNAs, Virus resistance
Citation
Wagaba, H., Patil, B. L., Mukasa, S., Alicai, T., Fauquet, C. M., & Taylor, N. J. (2016). Artificial microRNA-derived resistance to Cassava brown streak disease. Journal of virological methods, 231, 38-43. http://dx.doi.org/10.1016/j.jviromet.2016.02.004