KEY MESSAGE: In Physcomitrium patens, PpRH1/PpRH2 are GUCT-domain-containing DEAD-BOX RNA helicases localize to the nucleus. They are implicated in cell and tissue development in all stages of the moss life cycle. ABSTRACT: The DEAD-box-containing RNA helicase family encompasses a large and functionally important group of enzymes involved in cellular processes committed to the metabolism of RNA, including its transcription, processing, transport, translation and decay. Studies indicate this protein family has implied roles in plant vegetative and reproductive developmental processes as well as response to environmental stresses such has cold and high salinity. We focus here on a small conserved sub-group of GUCT domain-containing RNA helicase in the moss Physcomitrium patens. Phylogenetic analysis shows that RNA helicases containing the GUCT domain form a distinct conserved clade across the green lineage. In this clade, the P. patens genome possesses two closely related paralogues RNA helicases predicted to be nuclear, PpRH1 and PpRH2. Using in-locus gene fluorescent tagging we show that PpRH1 is localized to the nucleus in protonema. Analysis of PpRH1 and PpRH2 deletions, individually and together, indicates their potential roles in protonema, gametophore and sporophyte cellular and tissue development in P. patens. Additionally, the ultrastructural analysis of phyllid chloroplasts in Δrh2 and Δrh1/2 shows distinct starch granule accumulation under standard growth conditions associated with changes in photosynthetic activity parameters. We could not detect effects of either temperature or stress on protonema growth or PpRH1 and PpRH2 expression. Together, these results suggest that nuclear GUCT-containing RNA helicases play a role primarily in developmental processes directly or indirectly linked to photosynthesis activity in the moss P. patens. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s11103-021-01152-w.

Comenius University in Bratislava Science Park Ilkovicova 8 84215 Bratislava Slovakia

Department of Biophysics Centre of the Region Haná for Biotechnological and Agricultural Research Faculty of Science Palacký University Šlechtitelů 27 78371 Olomouc Czech Republic

Department of Biotechnology Inland Norway University of Applied Sciences Holsetgata 31 2318 Hamar Norway

Department of Gastroenterology Hepatology and Endocrinology Hannover Medical School Carl Neuberg Str 1 30625 Hannover Germany

Department of Plant Physiology Faculty of Natural Sciences Comenius University in Bratislava Ilkovicova 6 84215 Bratislava Slovakia

Institut Jean Pierre Bourgin INRAE AgroParisTech Université Paris Saclay 78000 Versailles France

Plant Cell Biology Faculty of Biology University of Marburg Karl von Frisch Str 8 35043 Marburg Germany

Plant Science and Biodiversity Center Slovak Academy of Sciences Dúbravská cesta 9 84523 Bratislava Slovakia

School of Animal Rural and Environmental Sciences Nottingham Trent University Brackenhurst Campus Southwell NG25 0QF Nottinghamshire UK

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