Differential miRNA profiles in South African cassava mosaic virus-infected cassava landraces reveal clues to susceptibility and tolerance to cassava mosaic disease.

Specific miRNA families are involved in susceptibility or antiviral immunity in plants. Manihot esculenta Crantz (cassava) is a perennial plant that is an important food security crop in sub-Saharan Africa. Cassava is susceptible to several begomoviruses that cause cassava mosaic disease (CMD). In this study, we investigated the leaf miRNAome response in a tolerant (TME3) and susceptible (T200) cassava landrace challenged with South African cassava mosaic virus. RNAseq was performed on leaf samples at 12, 32 and 67 days post infection (dpi), representing early, symptomatic and late persistent stages of CMD infection. Significantly, distinct profiles of conserved miRNA family expression between the T200 and TME3 landraces at the three infection stages were observed. Notably at 12 days post SACMV infection, TME3 exhibited significant downregulation (log2fold<2.0) of 42 %, compared to 9% in T200, of the conserved miRNA families. This demonstrates an overall early response to SACMV in TME3 prior to symptom appearance not observed in T200, and expression of a large cohort of miRNA-regulated genes. Notably, at early infection, downregulation of mes-miR162 and 168 that target antiviral posttransriptional gene silencing (PTGS) regulators DCL1 and AGO1, respectively, was observed in TME3, and AGO1 and DCL1 expression was higher compared to T200 post infection. Early rapid responses prior to symptom development, including RNA silencing, may be key to establishing the tolerance/recovery phenotype exhibited by TME3 landrace later on at 67 dpi. At recovery, TME3 was hallmarked by a highly significant down-regulation of mes-miR167. MiR167 targets an auxin responsive factor which plays a role in auxin signaling and adaptive responses to stress, suggesting the importance of the auxin signaling in recovery of SACMV-induced symptoms. The gene targets of these miRNAs and their associated networks may provide clues to the molecular basis of CMD tolerance in perennial hosts such as cassava.