Recent evidence has shown that microRNAs are transferred from one species to another through cross-species transmission and exhibit biological activities in the receptor. However, the cross-kingdom regulation of pathogen virulence by plant-derived miRNAs is rarely reported. This study investigated the regulatory role of novel tomato miRNA miR1001 in the growth and development of Botrytis cinerea. Results showed that miR1001 inhibited the virulence of B. cinerea-infected plants, and the inhibitory effect of miR1001/miR1001* was stronger than that of miR1001. Moreover, miR1001 exerted a significant inhibitory effect on the conidiospore germination of B. cinerea. Degradome-seq experiment showed that miR1001 can directly target the Bcin03g02170.1 and Bcin10g01400.1 genes, which respectively encode the ATP-dependent metallopeptidase and cysteine-type endopeptidase, in B. cinerea. The interactions of both targets with miR1001 were further confirmed by using transient co-expression in tobacco. Real-time RT-PCR analysis showed that the expression levels of the two target genes were significantly downregulated in B. cinerea with miR1001 treatment. Our findings provide new evidence into the coevolution of pathogens and host plants, as well as new directions for the use of plant-derived miRNAs to control pathogens.

最近的证据表明，microRNA通过种间传递从一种物种转移到另一种物种，并在受体中表现出生物活性。但是，很少有关于植物来源的miRNA对病原体毒力的跨界调节的报道。本研究调查了新型番茄miRNA miR1001在灰葡萄孢生长发育中的调控作用。结果表明，miR1001抑制了灰葡萄孢感染的植物的毒性，并且miR1001 / miR1001 *的抑制作用强于miR1001。而且，miR1001对灰质芽孢杆菌的分生孢子萌发具有显着的抑制作用。降解序列实验表明miR1001可以直接靶向Bcin03g02170.1和Bcin10g01400.1基因，分别编码B. cinerea中的ATP依赖性金属肽酶和半胱氨酸型内肽酶。通过在烟草中使用瞬时共表达，进一步证实了两个靶标与miR1001的相互作用。实时RT-PCR分析显示，miR1001处理可在灰葡萄孢中显着下调两个靶基因的表达水平。我们的发现为病原体和宿主植物的共同进化提供了新的证据，以及使用植物来源的miRNA控制病原体的新方向。