Sabine Zachgo’s research while affiliated with Osnabrück University and other places

Key message We link key aspects of land plant reproductive evolution and detail how successive molecular changes leading to novel tissues and organs require co-evolution of communication systems between tissues. Abstract The transition of water-dependent reproduction of algae to mechanisms with very limited water dependence in many land plant lineages allowed plants to colonize diverse terrestrial environments, leading to the vast variety of extant plant species. The emergence of modified cell types, novel tissues, and organs enabled this transition; their origin is associated with the co-evolution of novel or adapted molecular communication systems and gene regulatory networks. In the light of an increasing number of genome sequences in combination with the establishment of novel genetic model organisms from diverse green plant lineages, our knowledge and understanding about the origin and evolution of individual traits that arose in a concerted way increases steadily. For example, novel members of gene families in signaling pathways emerged for communication between gametes and gametophytes with additional tissues surrounding the gametes. Here, we provide a comprehensive overview on the origin and evolution of reproductive novelties such as pollen grains, immobile sperms, ovules and seeds, carpels, gamete/gametophytic communication systems, double fertilization, and the molecular mechanisms that have arisen anew or have been co-opted during evolution, including but not limited to the incorporation of phytohormones, reactive oxygen species and redox signaling as well as small RNAs in regulatory modules that contributed to the evolution of land plant sexual reproduction.

DICER‐LIKE (DCL) proteins play a central role in plant small RNA (sRNA) biogenesis. The genome of the early land plant Marchantia polymorpha encodes four DCL proteins: MpDCL1a, MpDCL1b, MpDCL3, and MpDCL4. While MpDCL1a, MpDCL3 and MpDCL4 show high similarities to their orthologs in Physcomitrium patens and Arabidopsis thaliana, MpDCL1b shares only a limited homology with PpDCL1b, but it is very similar, in terms of functional domains, to orthologs in other moss and fern species. We generated Mpdclge mutant lines for all MpDCL genes with the CRISPR/Cas9 system and conducted phenotypic analyses under control, salt stress, and phytohormone treatments to uncover specific MpDCL functions. The mutants displayed severe developmental aberrations, altered responses to salt and phytohormones, and disturbed sexual organ development. By combining mRNA and sRNA analyses, we demonstrate that MpDCLs and their associated sRNAs play pivotal roles in regulating development, abiotic stress tolerance and phytohormone response in M. polymorpha. We identified MpDCL1a in microRNA biogenesis, MpDCL4 in trans‐acting small interfering RNA generation, and MpDCL3 in the regulation of pathogen‐related genes. Notably, salt sensitivity in M. polymorpha is dependent on MpDCL1b and Mpdcl1bge mutants display enhanced tolerance and reduced miRNA expression in response to salt stress. We propose that M. polymorpha employs specific mechanisms for regulating MpDCL1b associated miRNAs under high salinity conditions, potentially shared with other species harboring MpDCL1b homologs.

Land plants evolved from an ancestral alga around 470 mya, evolving complex multicellularity in both haploid gametophyte and diploid sporophyte generations. The evolution of water-conducting tissues in the sporo- phyte generation was crucial for the success of land plants, paving the way for the colonization of a variety of terrestrial habitats. Class II KNOX (KNOX2) genes are major regulators of secondary cell wall formation and seed mucilage (pectin) deposition in flowering plants. Here, we show that, in the liverwort Marchantia poly- morpha, loss-of-function alleles of the KNOX2 ortholog, MpKNOX2, or its dimerization partner, MpBELL1, have defects in capsule wall secondary cell wall and spore pectin biosynthesis. Both genes are expressed in the gametophytic calyptra surrounding the sporophyte and exert maternal effects, suggesting intergener- ational regulation from the maternal gametophyte to the sporophytic embryo. These findings also suggest the presence of a secondary wall genetic program in the non-vascular liverwort capsule wall, with attributes of secondary walls in vascular tissues.

The NPR proteins function as salicylic acid (SA) receptors in Arabidopsis thaliana. AtNPR1 plays a central role in SA‐induced transcriptional reprogramming whereby positively regulates SA‐mediated defense. NPRs are found in the genomes of nearly all land plants. However, we know little about the molecular functions and physiological roles of NPRs in most plant species. We conducted phylogenetic and alignment analyses of NPRs from 68 species covering the significant lineages of land plants. To investigate NPR functions in bryophyte lineages, we generated and characterized NPR loss‐of‐function mutants in the liverwort Marchantia polymorpha. Brassicaceae NPR1‐like proteins have characteristically gained or lost functional residues identified in AtNPRs, pointing to the possibility of a unique evolutionary trajectory for the Brassicaceae NPR1‐like proteins. We find that the only NPR in M. polymorpha, MpNPR, is not the master regulator of SA‐induced transcriptional reprogramming and negatively regulates bacterial resistance in this species. The Mpnpr transcriptome suggested roles of MpNPR in heat and far‐red light responses. We identify both Mpnpr and Atnpr1‐1 display enhanced thermomorphogenesis. Interspecies complementation analysis indicated that the molecular properties of AtNPR1 and MpNPR are partially conserved. We further show that MpNPR has SA‐binding activity. NPRs and NPR‐associated pathways have evolved distinctively in diverged land plant lineages to cope with different terrestrial environments.

The plant cuticle is a hydrophobic barrier, which seals the epidermal surface of most aboveground organs. While the cuticle biosynthesis of angiosperms has been intensively studied, knowledge about its existence and composition in nonvascular plants is scarce. Here, we identified and characterized homologs of Arabidopsis thaliana fatty acyl‐CoA reductase (FAR) ECERIFERUM 4 (AtCER4) and bifunctional wax ester synthase/acyl‐CoA:diacylglycerol acyltransferase 1 (AtWSD1) in the liverwort Marchantia polymorpha (MpFAR2 and MpWSD1) and the moss Physcomitrium patens (PpFAR2A, PpFAR2B, and PpWSD1). Although bryophyte harbor similar compound classes as described for angiosperm cuticles, their biosynthesis may not be fully conserved between the bryophytes M. polymorpha and P. patens or between these bryophytes and angiosperms. While PpFAR2A and PpFAR2B contribute to the production of primary alcohols in P. patens, loss of MpFAR2 function does not affect the wax profile of M. polymorpha. By contrast, MpWSD1 acts as the major wax ester‐producing enzyme in M. polymorpha, whereas mutations of PpWSD1 do not affect the wax ester levels of P. patens. Our results suggest that the biosynthetic enzymes involved in primary alcohol and wax ester formation in land plants have either evolved multiple times independently or undergone pronounced radiation followed by the formation of lineage‐specific toolkits.

In angiosperms, basic leucine‐zipper (bZIP) TGACG‐motif‐binding (TGA) transcription factors (TFs) regulate developmental and stress‐related processes, the latter often involving NON EXPRESSOR OF PATHOGENESIS‐RELATED GENES (NPR) coregulator interactions. To gain insight into their functions in an early diverging land‐plant lineage, the single MpTGA and sole MpNPR genes were investigated in the liverwort Marchantia polymorpha. We generated Marchantia MpTGA and MpNPR knockout and overexpression mutants and conducted morphological, transcriptomic and expression studies. Furthermore, we investigated MpTGA interactions with wild‐type and mutagenized MpNPR and expanded our analyses including TGA TFs from two streptophyte algae. Mptga mutants fail to induce the switch from vegetative to reproductive development and lack gametangiophore formation. MpTGA and MpNPR proteins interact and Mpnpr mutant analysis reveals a novel coregulatory NPR role in sexual reproduction. Additionally, MpTGA acts independently of MpNPR as a repressor of oil body (OB) formation and can thereby affect herbivory. The single MpTGA TF exerts a dual role in sexual reproduction and OB formation in Marchantia. Common activities of MpTGA/MpNPR in sexual development suggest that coregulatory interactions were established after emergence of land‐plant‐specific NPR genes and contributed to the diversification of TGA TF functions during land‐plant evolution.

The bacterium Paenibacillus marchantiae was isolated from male plants of the liverwort Marchantia polymorpha subsp. ruderalis ecotype BoGa. Here, we report on the complete genome sequence generated from long Nanopore reads. The genome sequence comprises 6,983,959 bp with a GC content of 46.02% and 6,195 predicted protein-coding genes.

Here, we present the Nanopore-only genome sequence of Aneurinibacillus sp. Ricciae_BoGa-3. It was isolated from Riccia fluitans ecotype BoGa-3 and its source was Botanical Garden Osnabrück (Germany). The complete circular genome is 4,981,254 bp with a GC content of 44.8%.

Recent findings expanded our knowledge about plant redox regulation in stress responses by demonstrating that redox processes exert crucial nuclear regulatory functions in meristems and other developmental processes. Analyses of redox-modulated transcription factor functions and coregulatory ROXYs, CC-type land-plant specific glutaredoxins, reveal new insights into the redox control of plant transcription factors and participation of ROXYs in plant development. The role for ROS and redox signaling in response to low-oxygen conditions further strengthens the importance of redox processes in meristems and tissue differentiation as well as for adaptation to changing environments effecting food crop productivity.