Marchantia Polymorpha Research Articles

3D anatomy and dorsoventral asymmetry of the mature Marchantia polymorpha meristem develops from a symmetrical gemma meristem.

Meristems are three-dimensional generative structures that contain stem cells and produce new organs and tissues. Meristems develop in all land plants, however we know little about the spatial and temporal regulation of meristem structure in lineages such as bryophytes. Here we describe the three-dimensional meristem anatomy during the development of the liverwort, Marchantia polymorpha. We show that the apical stem cell of the mature meristem is sub-apical, ventral, and in the outer cell layer. Mature meristem anatomy is therefore asymmetrical in the dorsoventral axis, which is reflected by the domain specific protein localisation of Class III and Class IV Homeodomain-Leucine-Zippers (MpC3HDZ and MpC4HDZ) and promoter activity of MpYUCCA2. The dorsoventral asymmetry that defines the mature meristem is absent in the juvenile meristems of asexual propagules known as gemmae. We discovered that anatomical dorsoventral asymmetry of the meristem forms after one to two days of gemmaling growth and is accompanied by expression of the dorsal identity reporter, MpC3HDZ. We conclude that the gemma meristem has arrested development and undergoes anatomical rearrangement to develop the three-dimensional meristem structure of the mature plant.

MID1-COMPLEMENTING ACTIVITY regulates cell proliferation and development via Ca2+ signaling in Marchantia polymorpha.

MID1-COMPLEMENTING ACTIVITY (MCA) is a land plant-specific, plasma membrane protein and Ca2+ signaling component that responds to exogenous mechanical stimuli, such as touch, gravity, and hypotonic-osmotic stress, in various plant species. MCA is essential for cell proliferation and differentiation during growth and development in rice (Oryza sativa) and maize (Zea mays). However, the mechanism by which MCA mediates cell proliferation and differentiation via Ca2+ signaling remains unknown. Here, we address this question using the liverwort Marchantia polymorpha. We show that the M. polymorpha MCA ortholog, MpMCA, is highly expressed in actively dividing regions, such as apical notches in the thalli and developing gametangiophores, and that MpMCA is a plasma membrane protein. In vivo Ca2+ imaging using a Ca2+ sensor (yellow cameleon) revealed that MpMCA is required for maintaining proper [Ca2+]cyt levels in the apical notch region, egg cells, and antheridium cells. Mpmca mutant plants showed severe cell proliferation and differentiation defects in the thalli, gametangiophores, and gametangia, resulting in abnormal development and unsuccessful fertilization. Furthermore, expression of the Arabidopsis MCA1 gene complemented most of the defects in the growth and development of the Mpmca mutant plants. Our findings indicate that MpMCA is an evolutionarily conserved Ca2+-signaling component that regulates cell proliferation and development across the life cycle of land plants.

Inositol pyrophosphate catabolism by three families of phosphatases regulates plant growth and development.

Inositol pyrophosphates (PP-InsPs) are nutrient messengers whose cellular levels are precisely regulated. Diphosphoinositol pentakisphosphate kinases (PPIP5Ks) generate the active signaling molecule 1,5-InsP8. PPIP5Ks harbor phosphatase domains that hydrolyze PP-InsPs. Plant and Fungi Atypical Dual Specificity Phosphatases (PFA-DSPs) and NUDIX phosphatases (NUDTs) are also involved in PP-InsP degradation. Here, we analyze the relative contributions of the three different phosphatase families to plant PP-InsP catabolism. We report the biochemical characterization of inositol pyrophosphate phosphatases from Arabidopsis and Marchantia polymorpha. Overexpression of different PFA-DSP and NUDT enzymes affects PP-InsP levels and leads to stunted growth phenotypes in Arabidopsis. nudt17/18/21 knock-out mutants have altered PP-InsP pools and gene expression patterns, but no apparent growth defects. In contrast, Marchantia polymorpha Mppfa-dsp1ge, Mpnudt1ge and Mpvip1ge mutants display severe growth and developmental phenotypes and associated changes in cellular PP-InsP levels. Analysis of Mppfa-dsp1geand Mpvip1ge mutants supports a role for PP-InsPs in Marchantia phosphate signaling, and additional functions in nitrate homeostasis and cell wall biogenesis. Simultaneous elimination of two phosphatase activities enhanced the observed growth phenotypes. Taken together, PPIP5K, PFA-DSP and NUDT inositol pyrophosphate phosphatases regulate growth and development by collectively shaping plant PP-InsP pools.

MurA-catalyzed synthesis of 5-enolpyruvylshikimate-3-phosphate confers glyphosate tolerance in bryophytes

Glyphosate is a broad-spectrum herbicide that kills most vascular plant weeds but is ineffective against many bryophytes. Glyphosate competitively inhibits the enolpyruvyl transferase enzyme 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS). EPSPS catalyzes the production of 5-enolpyruvylshikimate-3-phosphate (EPSP)-an intermediate in the shikimate pathway-from shikimate-3-phosphate (S3P) and phosphoenolpyruvate (PEP) substrates. Here, we show that Marchantia polymorpha mutants with loss-of-function mutations in a closely related enolpyruvyl transferase, MpMurA, were more sensitive to glyphosate than wild-type controls. Overexpression of MpMurA in M. polymorpha increased glyphosate tolerance, and heterologous expression of the M. polymorpha MurA enzyme in Arabidopsis thaliana conferred glyphosate resistance. Furthermore, we demonstrate that MpMurA catalyzes the production of EPSP from S3P and PEP substrates. These data demonstrate that MpMurA contributes to glyphosate tolerance in M. polymorpha. We speculate that the existence of two independent mechanisms for EPSP synthesis-one canonical and EPSPS-dependent, and the other MurA-dependent-may account for glyphosate tolerance in bryophytes. Alternatively, MurA in bryophytes may bind glyphosate, thereby leaving more unbound EPSPS enzymes available, allowing aromatic amino acid biosynthesis to continue.

Dn-OPDA conjugation with amino acids inhibits its phytohormone bioactivity in Marchantia polymorpha.

Jasmonates are important phytohormones that regulate plant tolerance to biotic and abiotic stresses, and developmental processes. Distinct jasmonates in different plant lineages activate a conserved signalling pathway that mediates these responses: dinor-12-oxo-phytodienoic acid (dn-OPDA) isomers in bryophytes and lycophytes, and JA-Ile in most vascular plants. In many cases, the final responses triggered by these phytohormones depend on the accumulation of specialized metabolites. To identify compounds regulated by the dn-OPDA pathway in the liverwort Marchantia polymorpha, untargeted metabolomic analyses were carried out in response to wounding, a stress that activates the dn-OPDA pathway. A previously unreported group of molecules was identified from these analyses: dn-OPDA-amino acid conjugates (dn-OPDA-aas). Their accumulation after wounding and herbivory was confirmed by targeted metabolic profiling in Marchantia and in all species in which we previously detected dn-iso-OPDA. Mutants in GRETCHEN-HAGEN 3A (MpGH3A) failed to accumulate dn-OPDA-aa conjugates and showed a constitutive activation of the OPDA pathway and increased resistance to herbivory. Our results show that dn-iso-OPDA bioactivity is reduced by amino acid conjugation. Therefore, jasmonate conjugation in land plants plays dichotomous roles: jasmonic acid (JA) conjugation with isoleucine (Ile) produces the bioactive JA-Ile in tracheophytes, whereas conjugation of dn-iso-OPDA with different amino acids deactivates the phytohormone in bryophytes and lycophytes.

Environmental Pollutant Anthracene Induces ABA-Dependent Transgenerational Effects on Gemmae Dormancy in Marchantia polymorpha.

Anthracene, a polycyclic aromatic hydrocarbon (PAH) from fossil fuel combustion, poses significant environmental threats. This study investigates the role of abscisic acid (ABA) in the anthracene tolerance of the liverwort Marchantia polymorpha using mutants deficient in ABA perception (Mppyl1) or biosynthesis (Mpaba1). In this study, we monitored the role of ABA in the anthracene tolerance response by tracking two ABA-controlled traits: plant growth inhibition and gemmae dormancy. We found that the anthracene-induced inhibition of plant growth is dose-dependent, similar to the growth-inhibiting effect of ABA, but independent of ABA pathways. However, gemmae dormancy was differentially affected by anthracene in ABA-deficient mutants. We found that gemmae from anthracene-exposed WT plants exhibited reduced germination compared to those from mock-treated plants. This suggests that the anthracene exposure of mother plants induces a transgenerational effect, resulting in prolonged dormancy in their asexual propagules. While Mppyl1 gemmae retained a dormancy delay when derived from anthracene-exposed thalli, the ABA biosynthesis mutant Mpaba1 did not display any significant dormancy delay as a consequence of anthracene exposure. These results, together with the strong induction of ABA marker genes upon anthracene treatment, imply that anthracene-induced germination inhibition relies on ABA synthesis in the mother plant, highlighting the critical role of MpABA1 in the tolerance response. These findings reveal a complex interplay between anthracene stress and ABA signaling, where anthracene triggers ABA-mediated responses, influencing reproductive success and highlighting the potential for leveraging genetic and hormonal pathways to enhance plant resilience in contaminated habitats.

Functional Conservation of the DDP1-type Inositol Pyrophosphate Phosphohydrolases in Land Plant.

Inositol pyrophosphates (PP-InsPs) are eukaryote-specific second messengers that regulate diverse cellular processes, including immunity, nutrient sensing, and hormone signaling pathways in plants. These energy-rich messengers exhibit high sensitivity to the cellular phosphate status, suggesting that the synthesis and degradation of PP-InsPs are tightly controlled within the cells. Notably, the molecular basis of PP-InsP hydrolysis in plants remains largely unexplored. In this study, we report the functional characterization of MpDDP1, a diadenosine and diphosphoinositol polyphosphate phosphohydrolase encoded by the genome of the liverwort, Marchantia polymorpha. We show that MpDDP1 functions as a PP-InsP phosphohydrolase in different heterologous organisms. Consistent with this finding, M. polymorpha plants defective in MpDDP1 exhibit elevated levels of 1/3-InsP7 and 1/3,5-InsP8, highlighting the contribution of MpDDP1 in regulating PP-InsP homeostasis in planta. Furthermore, our study reveals that MpDDP1 controls thallus development and vegetative reproduction in M. polymorpha. Collectively, this study provides insights into the regulation of specific PP-InsP messengers by DDP1-type phosphohydrolases in land plants.

Microtubules and actin filaments direct nuclear movement during the polarisation of Marchantia spore cells.

The multicellular haploid stage of land plants develops from a single haploid cell produced by meiosis - the spore. Starting from a non-polar state, these spores develop polarity, divide asymmetrically and establish the first axis of symmetry. Here, we show that the nucleus migrates from the cell centroid to the basal pole during polarisation of the Marchantia polymorpha spore cell. A microtubule organising centre on the leading edge of the nucleus initiates a microtubule array between the nuclear surface and the cortex at the basal pole. Simultaneously, cortical microtubules disappear from the apical hemisphere but persist in the basal hemisphere. This is accompanied by the formation a dense network of fine actin filaments between the nucleus and the basal pole cortex. Experimental depolymerisation of either microtubules or actin filaments disrupts cellular asymmetry. These data demonstrate that the cytoskeleton reorganises during spore polarisation and controls the directed migration of the nucleus to the basal pole. The presence of the nucleus at the basal pole provides the cellular asymmetry for the asymmetric cell division that establishes the apical-basal axis of the plant.

An interview with Liam Dolan.

Liam Dolan is a Group Leader at the Gregor Mendel Institute of Molecular Plant Biology (GMI) in Vienna, Austria, interested in intersection of plant development, evolution and ecology using the emerging model system Marchantia polymorpha. We met with Liam at the GMI to learn more about his career so far, his views on biodiversity and the ecosystem, and why he became involved as a Guest Editor for Development's Special Issue: Uncovering Developmental Diversity.

Evolutionary Conserved and Divergent Responses to Copper Zinc Superoxide Dismutase Inhibition in Plants.

After an initial evolution in a reducing environment, life got successively challenged by reactive oxygen species (ROS), especially during the great oxidation event (GOE) that followed the development of photosynthesis. Therefore, ROS are deeply intertwined into the physiological, morphological and transcriptional responses of most present-day organisms. Copper-zinc superoxide dismutases (CuZnSODs) evolved during the GOE and are present in charophytes and extant land plants, but nearly absent from chlorophytes. The chemical inhibitor of CuZnSOD, lung cancer screen 1 (LCS-1), could greatly facilitate the study of SODs in diverse plants. Here, we determined the impact of chemical inhibition of plant CuZnSOD activity, on plant growth, transcription and metabolism. We followed a comparative approach by using different plant species, including Marchantia Polymorpha and Physcomitrium patens, representing bryophytes, the sister lineage to vascular plants, and Arabidopsis thaliana. We show that LCS-1 causes oxidative stress in plants and that the inhibition of CuZnSODs provoked a similar core response that mainly impacted glutathione homoeostasis in all plant species analysed. That said, Physcomitrium and Arabidopsis, which contain multiple CuZnSOD isoforms showed a more complex and exacerbated response. In addition, an untargeted metabolomics approach revealed a specific metabolic signature for each plant species. Our comparative analysis exposes a conserved core response at the physiological and transcriptional level towards LCS-1, while the metabolic response largely varies. These differences correlate with the number and localization of the CuZnSOD isoforms present in each species.

MpPUB9, a U-box E3 ubiquitin ligase, acts as a positive regulator by promoting the turnover of MpEXO70.1 under high salinity in Marchantia polymorpha.

Marchantia polymorpha, occupying a basal position in the monophyletic assemblage of land plants, displays a notable expansion of plant U-box (PUB) proteins compared with those in animals. We elucidated the roles of MpPUB9 in regulating salt stress tolerance in M. polymorpha. MpPUB9 expression was rapidly induced by high salinity and dehydration. MpPUB9 possessed an intact U-box domain in the N-terminus. MpPUB9-Citrine localized to punctate structures and was peripherally associated with microsomal membranes. Phenotypic analyses demonstrate that the hyponastic and epinastic thallus growth phenotypes, which were induced by the overexpression and suppression of MpPUB9, may provoke salt stress-resistant and -susceptible phenotypes, respectively. MpPUB9 was also found to directly interact with the exocyst protein MpEXO70.1, leading to its ubiquitination. Under high-salinity conditions, though the stability of MpPUB9 was dramatically increased, MpEXO70.1 showed slightly faster turnover rates. Transcriptome analyses showed that salt treatment and the overexpression of MpPUB9 co-upregulated the genes related to the modulation of H2O2 and cell wall organization. Overall, our results suggest that MpPUB9 plays a crucial role in the positive regulation of salt stress tolerance, resulting from its interaction with MpEXO70.1 and modulating turnover of the protein under high-salt conditions via the coordination of UPS with autophagy.

Real-time live imaging of cytosolic hydrogen peroxide and Ca2+ of Marchantia polymorpha gemmalings reveal immediate initial responses of plant cells triggered by nonthermal plasma irradiation

Cold atmospheric pressure plasma generators capable of generating plasma under normal pressure and temperature conditions have recently been developed, and their biological applications have been extensively studied. Plasma irradiation has been reported to affect plant germination and growth; however, the molecular mechanism underlying these effects and initial cellular responses to plasma irradiation remains poorly understood. To unravel the molecular and cellular mechanisms underlying the effects of plasma irradiation on plants, we have been establishing novel experimental systems using a model liverwort Marchantia polymorpha. We here focused on the initial responses of plant cells to plasma irradiation. To investigate immediate cellular responses following plasma irradiation, we developed a new plasma device that allows irradiation under a microscope. Through integration with live fluorescence imaging, we established an experimental setup to track, the dynamics of intracellular concentration of H2O2 and Ca2+ as representative initial cellular responses. We revealed that plasma irradiation induced a rapid and transient increase in intracellular concentration of H2O2 and Ca2+ in Marchantia gemmalings. Pharmacological analyses suggested that the long-lived reactive species, H2O2, generated by the plasma generator was directly delivered into the plant cells. Competitive inhibitors of Ca2+ channels abolished the Ca2+ rise, suggesting that plasma irradiation immediately activate plasma membrane Ca2+ channel(s) to induce Ca2+ influx. Importantly, this study marks the inaugural demonstration of real-time monitoring of cytosolic H2O2 and Ca2+ dynamics in plants, triggered by plasma irradiation.

Control of sporophyte secondary cell wall development in Marchantia by a Class II KNOX gene.

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 sporophyte 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 polymorpha, 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 intergenerational 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.

Conservation of molecular responses upon viral infection in the non-vascular plant Marchantia polymorpha

After plants transitioned from water to land around 450 million years ago, they faced novel pathogenic microbes. Their colonization of diverse habitats was driven by anatomical innovations like roots, stomata, and vascular tissue, which became central to plant-microbe interactions. However, the impact of these innovations on plant immunity and pathogen infection strategies remains poorly understood. Here, we explore plant-virus interactions in the bryophyte Marchantia polymorpha to gain insights into the evolution of these relationships. Virome analysis reveals that Marchantia is predominantly associated with RNA viruses. Comparative studies with tobacco mosaic virus (TMV) show that Marchantia shares core defense responses with vascular plants but also exhibits unique features, such as a sustained wound response preventing viral spread. Additionally, general defense responses in Marchantia are equivalent to those restricted to vascular tissues in Nicotiana, suggesting that evolutionary acquisition of developmental innovations results in re-routing of defense responses in vascular plants.

The auronidin flavonoid pigments of the liverwort Marchantia polymorpha form polymers that modify cell wall properties.

Plant adaptation from aquatic to terrestrial environments required modifications to cell wall structure and function to provide tolerance to new abiotic and biotic stressors. Here, we investigate the nature and function of red auronidin pigment accumulation in the cell wall of the liverwort Marchantia polymorpha. Transgenic plants with auronidin production either constitutive or absent were analysed for their cell wall properties, including fractionation of polysaccharide and phenolic components. While small amounts of auronidin and other flavonoids were loosely associated with the cell wall, the majority of the pigments were tightly associated, similar to what is observed in angiosperms for polyphenolics such as lignin. No evidence of covalent binding to a polysaccharide component was found: we propose auronidin is present in the wall as a physically entrapped large molecular weight polymer. The results suggested auronidin is a dual function molecule that can both screen excess light and increase wall strength, hydrophobicity and resistance to enzymatic degradation by pathogens. Thus, liverworts have expanded the core phenylpropanoid toolkit that was present in the ancestor of all land plants, to deliver a lineage-specific solution to some of the environmental stresses faced from a terrestrial lifestyle.

Collaboration between DELLA and MED15 to regulate transcription in plants.

Gibberellins (GA) are diterpenoids that are categorized as one of main hormones that promote major developmental responses such as germination and stem elongation in plants. DELLA proteins act as the key repressors of GA responses. They interact with hundreds of different proteins. While the functioning of DELLA as transcriptional coactivators has also been reported earlier, the actual mechanism still remains elusive. One recent report describes interaction of DELLA with Mediator subunit MED15 as one of the mechanisms contributing to its transcription activation capability (Hernández-García et al. 2024). Interestingly, this mechanism of DELLA-MED15 module-mediated transcription regulation seems to be very ancient conserved from bryophyte Marchantia polymorpha to dicot Arabidopsis thaliana.

Regulation of ROP GTPase cycling between active and inactive states is essential for vegetative organogenesis in Marchantia polymorpha.

Rho/Rac of plant (ROP) GTPases are plant-specific proteins that function as molecular switches, activated by guanine nucleotide exchange factors (GEFs) and inactivated by GTPase-activating proteins (GAPs). The bryophyte Marchantia polymorpha contains single copies of ROP (MpROP), GEFs [ROPGEF and SPIKE (SPK)] and GAPs [ROPGAP and ROP ENHANCER (REN)]. MpROP regulates the development of various tissues and organs, such as rhizoids, gemmae and air chambers. The ROPGEF KARAPPO (MpKAR) is essential for gemma initiation, but the functions of other ROP regulatory factors are less understood. This study focused on two GAPs: MpROPGAP and MpREN. Mpren single mutants showed defects in thallus growth, rhizoid tip growth, gemma development, and air-chamber formation, whereas Mpropgap mutants showed no visible abnormalities. However, Mpropgap Mpren double mutants had more severe phenotypes than the Mpren single mutants, suggesting backup roles of MpROPGAP in processes involving MpREN. Overexpression of MpROPGAP and MpREN resulted in similar gametophyte defects, highlighting the importance of MpROP activation/inactivation cycling (or balancing). Thus, MpREN predominantly, and MpROPGAP as a backup, regulate gametophyte development, likely by controlling MpROP activation in M. polymorpha.

The BNB-GLID module regulates germline fate determination in Marchantia polymorpha.

Germline fate determination is a critical event in sexual reproduction. Unlike animals, plants specify the germline by reprogramming somatic cells at the late stages of their development. However, the genetic basis of germline fate determination and how it evolved during the land plant evolution are still poorly understood. Here, we report that the plant homeodomain finger protein GERMLINE IDENTITY DETERMINANT (GLID) is a key regulator of the germline specification in liverwort, Marchantia polymorpha. Loss of the MpGLID function causes failure of germline initiation, leading to the absence of sperm and egg cells. Remarkably, the overexpression of MpGLID in M. polymorpha induces the ectopic formation of cells with male germline cell features exclusively in male thalli. We further show that MpBONOBO (BNB), with an evolutionarily conserved function, can induce the formation of male germ cell-like cells through the activation of MpGLID by directly binding to its promoter. The Arabidopsis (Arabidopsis thaliana) MpGLID ortholog, MALE STERILITY1 (AtMS1), fails to replace the germline specification function of MpGLID in M. polymorpha, demonstrating that a derived function of MpGLID orthologs has been restricted to tapetum development in flowering plants. Collectively, our findings suggest the presence of the BNB-GLID module in complex ancestral land plants that has been retained in bryophytes, but rewired in flowering plants for male germline fate determination.

Assessing the effects of allelopathic properties of organic mulches on liverwort control in container-grown ornamentals

Liverwort (Marchantia polymorpha) is a problematic weed in ornamental crop production. The major limitation of liverwort control is lack of herbicide options inside greenhouses and their potential to cause injury. The current research was undertaken to study the effectiveness of allelopathic properties of six different organic mulches including rice hull (RH), cocoa hull (CH), pine bark (PB), maple leaf (ML), shredded cypress (SC), and red hardwood (HW) for liverwort control. Mulch extracts were prepared and used to impregnate agar media at an increasing dose at 1X (2 mL mulch extract), 2X (4 mL), 3X (6 mL), and 4X (8 mL) rates, in a randomized complete block design. Ten liverwort gemmae were transferred to each petri dish and they were maintained inside a growth chamber. The number of gemmae germinating in each petri dish after 1 week and gemmae surviving at the end of 2 weeks was recorded. In greenhouse study, mulch extracts were applied to containers filled with substrate for assessing liverwort control. The mulch extracts or no extract (control) were applied to the containers uniformly at 1X (15 mL), 2X (30 mL), 3X (45 mL) or 4X (60 mL) rates, in a randomized complete block design. The percentage of substrate surface covered by liverwort thalli was visually estimated bi-weekly for 10 weeks. Fresh biomass of the thalli and gemmae cup counts in each pot were also recorded. After 1 week in the growth chamber, ML followed by SC, PB and RH extracts showed maximum suppression of liverwort gemmae germination. At 2 weeks, ML applied at either of the rates provided complete inhibition of liverwort growth. In greenhouse, all the mulch extracts were able to provide complete liverwort control for the first two weeks. All the mulches and rates of applications were significantly different from the control after 6, 8, and 10 weeks. PB and HW mulches showed excellent liverwort control and minimum fresh biomass of liverwort after 10 weeks as compared to other mulches. The allelopathic potential of the organic mulches can be a promising option for biopesticidal control of liverwort and a component of integrated liverwort management.

Diurnal.plant.tools in 2024: expanding to Marchantia polymorpha and four angiosperms.

Diurnal gene expression is a pervasive phenomenon occurring across all kingdoms of life, orchestrating adaptive responses to daily environmental fluctuations and thus enhancing organismal fitness. Our understanding of the plant circadian clock is primarily derived from studies in Arabidopsis and direct comparisons are difficult due to differences in gene family sizes. To this end, the identification of functional orthologs based on diurnal and tissue expression is necessary. The diurnal.plant.tools database constitutes a repository of gene expression profiles from 17 members of the Archaeplastida lineage, with built-in tools facilitating cross-species comparisons. In this database update, we expand the dataset with diurnal gene expression from 4 agriculturally significant crop species and Marchantia, a plant of evolutionary significance. Notably, the inclusion of diurnal gene expression data for Marchantia enables researchers to glean insights into the evolutionary trajectories of the circadian clock and other biological processes spanning from algae to angiosperms. Moreover, integrating diurnal gene expression data with datasets from related gene co-expression databases, such as CoNekt-Plants and CoNekt-Stress, which contain gene expression data for tissue and perturbation experiments, provides a comprehensive overview of gene functions across diverse biological contexts. This expanded database serves as a valuable resource for elucidating the intricacies of diurnal gene regulation and its evolutionary underpinnings in plant biology.