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Glomerella leaf spot (GLS) of apple ( Malus × domestica  Borkh.), caused by  Glomerella cingulata , is an emerging fungal epidemic threatening the apple industry. Little is known about the molecular mechanism underlying resistance to this devastating fungus. In this study, high-throughput sequencing technology was used to identify microRNAs (miRNAs) involved in GLS resistance in apple. We focused on miRNAs that target genes related to disease and found that expression of a novel miRNA,  Md-miRln20 , was higher in susceptible apple varieties than in resistant ones. Furthermore, its target gene  Md-TN1-GLS  exhibited the opposite expression pattern, which suggested that the expression levels of  Md-miRln20  and its target gene are closely related to apple resistance to GLS. Furthermore, downregulation of  Md-miRln20  in susceptible apple leaves resulted in upregulation of  Md-TN1-GLS  and reduced the disease incidence. Conversely, overexpression of  Md-miRln20  in resistant apple leaves suppressed  Md-TN1-GLS  expression, with increased disease incidence. We demonstrated that  Md-miRln20  negatively regulates resistance to GLS by suppressing  Md-TN1-GLS  expression and showed, for the first time, a crucial role for miRNA in response to GLS in apple.

A novel miRNA negatively regulates resistance to Glomerella leaf spot by suppressing expression of an NBS gene in apple