Artificial microRNA-derived resistance to Cassava brown streak disease

• DOI: 10.1016/j.jviromet.2016.02.004
• Publication Date: 2016-02-20T02:59:20Z
• Authors: Henry Wagaba; Basavaprabhu L. P...; Settumba Mukasa; Titus Alicai; Claude M. Fauquet; Nigel J. Taylor
• Keywords: Nicotiana; 0301 basic medicine; disease control; Short Communication; Ugandan cassava brown streak virus; control de enfermedades; virus; cassava; resistance; Artificial microRNAs; 03 medical and health sciences; Virus resistance; Virology; viruses; Cassava brown streak virus; resistencia; Disease Resistance; Plant Diseases; rna sequence; cassava brown streak disease; cassava brown streak virus; Cassava brown streak disease; manihot esculenta; plant viruses; virus de las plantas; Plants, Genetically Modified; 3. Good health; MicroRNAs; genomas; genomes; secuencia de arn

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 21nt 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.