show Abstracthide AbstractLong noncoding RNA (lncRNA) plays important roles in morphological differentiation and development in eukaryotes. In filamentous fungi, however, little is known about lncRNAs and their roles in sexual development. Here we describe sexual stage-induced lncRNAs during the formation of perithecium, the sexual fruiting body of Fusarium graminearum. We identified 547 lncRNAs whose expression were developmental stage-specific; about 40% of them peaked during ascus development, when meiosis occurs. A large fraction of the lncRNAs were found to be antisense to mRNAs, forming 300 sense–antisense pairs. Although small RNAs were produced from these overlapped loci, most of the antisense lncRNAs appeared not to be involved in gene silencing pathways. Rather, the expression of antisense lncRNA and sense mRNA pairs tended to be induced in parallel as the perithecium matured. To identify regulatory components for lncRNA expression, we analyzed mutants defective in the nonsense-mediated decay (NMD) pathway. A subset of lncRNAs was specifically targeted by NMD, suggesting a suppressive role of NMD in lncRNA expression during vegetative growth. This study provides comprehensive resources for studying developmental lncRNA that may be important for laying out the multicellular fruiting body plan. Overall design: Time-series mRNA profiles during the perithecial development for the Fusarium graminearum wild-type strain PH-1 were generated by deep sequencing, in triplicate, using the Illumina HiSeq 2500 system. Also, sexual_stage0 and sexual_stage4 data for xrn1-deletion mutant (?xrn1) were generated using the Illumina HiSeq 4000 system.