Non-coding RNAs play crucial roles in plant responses to viral stresses. However, their molecular mechanisms in tea leaf spot responses remain unclear. In this study, using Camellia sinensis, we identified lncRNA81246 as a long non-coding RNA that localizes to both the nucleus and cytoplasm. It functions as a competitive endogenous RNA, thereby disrupting CsNAC1 (encoding NAC domain-containing protein 1) degradation mediated by miR164d. Silencing lncRNA81246 increased the resistance of tea plants to presistanceathogens, whereas transient lncRNA81246-overexpression plants showed decreased resistance to pathogens. Co-expression assays in Nicotiana benthamiana revealed that lncRNA81246 affects the miR164d-CsNAC1 regulatory module. Transient miR164d-overexpression and silencing assays demonstrated its positive regulation of tea plant resistance. Specifically, silencing its target, CsNAC1,enhanced disease resistance, whereas transient overexpression reduced plant resistance. Yeast one-hybrid, dual-luciferase, and RT-qPCR assay results suggested that CsNAC1 alters the expression of CsEXLB1, whereas AsODN and tobacco transient overexpression assays showed that CsEXLB1 negatively regulated tea plant resistance. Thus, our research demonstrated that lncRNA81246 acts as a mediator to interfere with the miR164d-CsNAC1 regulatory module involved in the disease resistance of tea plants.
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