DICER-LIKE 5 loss causes thermosensitive male sterility in durum wheat and reveals an AU-rich motif guiding 24-nt phasiRNA biogenesis

• 成果类型: Article
• 署名作者: Belanger, Sebastien; Martin, Azahara C.; Marchant, D. Blaine; Zhan, Junpeng; McGregor, Madison; Smedley, Mark; Hayta, Sadiye; Moore, Graham; Meyers, Blake C.
• 署名单位: Donald Danforth Plant Science Center; James Hutton Institute; UK Research & Innovation (UKRI); Biotechnology and Biological Sciences Research Council (BBSRC); John Innes Center; Consejo Superior de Investigaciones Cientificas (CSIC); CSIC - Instituto de Agricultura Sostenible (IAS); University of Missouri System; University of Missouri Saint Louis; Huazhong Agricultural University; Huazhong Agricultural University; Hubei Hongshan Laboratory; University of California System; University of California Davis; University of California System; University of California Davis
• 刊物名称: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• ISSN/ISSBN: 0027-12662
• DOI: 10.1073/pnas.2504349122
• 发表日期: 2025-08-05
• 关键词: sensitive male-sterility; expression analysis; hybrid rice; rna; stringtie; alignment; barley; tools; hisat; lead

摘要

Reproductive, male-enriched small RNAs are present in flowering plants and animals, yet their role in plants remains underexplored. We generated dicer-like 5 (dcl5) mutants in durum wheat (Triticum turgidum ssp. durum 2n = 4x = 28; AABB), revealing temperature-sensitive genic male sterility. Loss of DCL5 depleted premeiotic and meiotic 24-nt phasiRNA production, correlating with sterility under standard growth conditions and partial fertility recovery at higher temperatures. A single functional DCL5 allele restored complete fertility, presenting a promising alternative to current methods for hybrid production. We demonstrate that premeiotic 24-nt phasiRNA biogenesis is independent of miRNA-mediated cleavage and driven by a conserved motif initiating DCL5 activity. In the dcl5 mutant developing under sterility-inducing conditions, developmental defects are observed during pollen maturation, rather than at peak 24-nt phasiRNA accumulation in premeiotic and meiotic anthers. Although no visible morphological abnormalities were apparent during early development, single-cell RNA sequencing revealed that dcl5 mutant cells exhibit transcriptional profiles distinct from those of wild-type cells, when premeiotic 24-nt phasiRNAs are accumulating at the early developmental stage. Finally, the coexpression ofArgonaute (AGO1b, AGO4a, and AGO6) homeologs in 24-nt phasiRNA-producing cells identifies candidate effectors and suggests a role for 24-nt phasiRNAs in transcriptional gene silencing.