Long non‐coding RNA Cslnc256 regulates tea plant resistance to anthracnose by suppressing CsmiR395 ‐mediated sulfate metabolism

SUMMARY Long non‐coding RNAs (lncRNAs) are critical regulators of stress responses in plants. Fungal pathogens such as Colletotrichum camelliae severely impair the development of tea plants ( Camellia sinensis ); however, mechanisms involving long ncRNAs (lncRNAs) acting as competing endogenous RNAs (ceRNAs) remain poorly understood in this pathosystem. Through transcriptome profiling of tea leaves 6 days post‐pathogen inoculation, a ceRNA regulatory network was constructed based on expression correlation analysis. A lncRNA localized in both the nucleus and cytoplasm, Cslnc256 , was identified to function as a molecular decoy for CsmiR395 , thereby protecting the sulfate transporter gene CsSULTR2;1 from CsmiR395 ‐mediated degradation. Our findings revealed that CsmiR395 ‐directed cleavage of CsSULTR2;1 positively regulated sulfate metabolism and enhanced disease resistance. Silencing Cslnc256 enhanced pathogen resistance, whereas transient overexpression reduced the plant defense capacity. Single‐base substitution mapping, coupled with Nicotiana benthamiana transient expression and β‐glucuronidase reporter assays, confirmed that the 1345–1356 bp region of Cslnc256 constitutes the critical interaction domain for CsmiR395 . This study elucidated the molecular mechanism by which the Cslnc256–CsmiR395–CsSULTR2;1 module dynamically regulates sulfur metabolism to coordinate tea plant responses, providing novel insights into RNA‐mediated regulatory networks that govern plant–pathogen interactions. These findings offer a new perspective for deciphering RNA‐layered responses in crop protection strategies.