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1.
Planta ; 260(2): 45, 2024 Jul 04.
Artigo em Inglês | MEDLINE | ID: mdl-38965075

RESUMO

MAIN CONCLUSION: Developing bryophytes differentially modify their plasmodesmata structure and function. Secondary plasmodesmata formation via twinning appears to be an ancestral trait. Plasmodesmata networks in hornwort sporophyte meristems resemble those of angiosperms. All land-plant taxa use plasmodesmata (PD) cell connections for symplasmic communication. In angiosperm development, PD networks undergo an extensive remodeling by structural and functional PD modifications, and by postcytokinetic formation of additional secondary PD (secPD). Since comparable information on PD dynamics is scarce for the embryophyte sister groups, we investigated maturating tissues of Anthoceros agrestis (hornwort), Physcomitrium patens (moss), and Marchantia polymorpha (liverwort). As in angiosperms, quantitative electron microscopy revealed secPD formation via twinning in gametophytes of all model bryophytes, which gives rise to laterally adjacent PD pairs or to complex branched PD. This finding suggests that PD twinning is an ancient evolutionary mechanism to adjust PD numbers during wall expansion. Moreover, all bryophyte gametophytes modify their existing PD via taxon-specific strategies resembling those of angiosperms. Development of type II-like PD morphotypes with enlarged diameters or formation of pit pairs might be required to maintain PD transport rates during wall thickening. Similar to angiosperm leaves, fluorescence redistribution after photobleaching revealed a considerable reduction of the PD permeability in maturating P. patens phyllids. In contrast to previous reports on monoplex meristems of bryophyte gametophytes with single initials, we observed targeted secPD formation in the multi-initial basal meristems of A. agrestis sporophytes. Their PD networks share typical features of multi-initial angiosperm meristems, which may hint at a putative homologous origin. We also discuss that monoplex and multi-initial meristems may require distinct types of PD networks, with or without secPD formation, to control maintenance of initial identity and positional signaling.


Assuntos
Plasmodesmos , Plasmodesmos/ultraestrutura , Plasmodesmos/metabolismo , Briófitas/crescimento & desenvolvimento , Briófitas/fisiologia , Briófitas/ultraestrutura , Bryopsida/crescimento & desenvolvimento , Bryopsida/fisiologia , Bryopsida/ultraestrutura , Marchantia/genética , Marchantia/crescimento & desenvolvimento , Marchantia/fisiologia , Marchantia/ultraestrutura , Células Germinativas Vegetais/crescimento & desenvolvimento , Anthocerotophyta/fisiologia , Anthocerotophyta/metabolismo , Meristema/crescimento & desenvolvimento , Meristema/ultraestrutura , Meristema/fisiologia
2.
Nat Plants ; 10(6): 848-856, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38831044

RESUMO

The de novo synthesis of genomes has made unprecedented progress and achieved milestones, particularly in bacteria and yeast. However, the process of synthesizing a multicellular plant genome has not progressed at the same pace, due to the complexity of multicellular plant genomes, technical difficulties associated with large genome size and structure, and the intricacies of gene regulation and expression in plants. Here we outline the bottom-up design principles for the de novo synthesis of the Physcomitrium patens (that is, earthmoss) genome. To facilitate international collaboration and accessibility, we have developed and launched a public online design platform called GenoDesigner. This platform offers an intuitive graphical interface enabling users to efficiently manipulate extensive genome sequences, even up to the gigabase level. This tool is poised to greatly expedite the synthesis of the P. patens genome, offering an essential reference and roadmap for the synthesis of plant genomes.


Assuntos
Bryopsida , Genoma de Planta , Bryopsida/genética , Biologia Sintética/métodos , Software
3.
Plant J ; 119(2): 895-915, 2024 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-38753873

RESUMO

Plant mitochondrial and chloroplast transcripts are subject to numerous events of specific cytidine-to-uridine (C-to-U) RNA editing to correct genetic information. Key protein factors for this process are specific RNA-binding pentatricopeptide repeat (PPR) proteins, which are encoded in the nucleus and post-translationally imported into the two endosymbiotic organelles. Despite hundreds of C-to-U editing sites in the plant organelles, no comparable editing has been found for nucleo-cytosolic mRNAs raising the question why plant RNA editing is restricted to chloroplasts and mitochondria. Here, we addressed this issue in the model moss Physcomitrium patens, where all PPR-type RNA editing factors comprise specific RNA-binding and cytidine deamination functionalities in single proteins. To explore whether organelle-type RNA editing can principally also take place in the plant cytosol, we expressed PPR56, PPR65 and PPR78, three editing factors recently shown to also function in a bacterial setup, together with cytosolic co-transcribed native targets in Physcomitrium. While we obtained unsatisfying results upon their constitutive expression, we found strong cytosolic RNA editing under hormone-inducible expression. Moreover, RNA-Seq analyses revealed varying numbers of up to more than 900 off-targets in other cytosolic transcripts. We conclude that PPR-mediated C-to-U RNA editing is not per se incompatible with the plant cytosol but that its limited target specificity has restricted its occurrence to the much less complex transcriptomes of mitochondria and chloroplast in the course of evolution.


Assuntos
Bryopsida , Cloroplastos , Citosol , Mitocôndrias , Edição de RNA , RNA de Plantas , Cloroplastos/metabolismo , Cloroplastos/genética , Citosol/metabolismo , Bryopsida/genética , Bryopsida/metabolismo , Mitocôndrias/metabolismo , Mitocôndrias/genética , RNA de Plantas/genética , RNA de Plantas/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Citidina/metabolismo , Citidina/genética , Proteínas de Ligação a RNA/metabolismo , Proteínas de Ligação a RNA/genética , Regulação da Expressão Gênica de Plantas , Uridina/metabolismo , Uridina/genética
4.
Ann Bot ; 134(2): 351-364, 2024 Jul 09.
Artigo em Inglês | MEDLINE | ID: mdl-38702965

RESUMO

BACKGROUND AND AIMS: Desiccation tolerance (DT) is crucial for survival in arid environments, where organisms develop strategies in reproduction, maintenance and defence to cope with water scarcity. Therefore, investigating the relationship between reproduction and DT is essential to understand the ecology and adaptive strategies of species. This study explores the connection between the development of male and female gametangia in the moss Bryum argenteum and the decrease in DT during the progression of phenological phases in gametangia and protonema. METHODS: Samples collected from a dry tropical forest in Brazil were cultivated, cloned and subjected to desiccation. Subsequently, the physiological parameters of shoots and protonemata were analysed. Shoot and protonema regeneration were monitored for 28 d after the physiological analyses. Both phases were subjected to control and desiccation treatments. KEY RESULTS: Significant effects of desiccation and sex on the physiological parameters and regeneration capacity of shoots and protonemata were found. Male shoots generally exhibited lower values of Fv/Fm (quantum efficiency of photosystem II) and ϕPSII (effective quantum yield of photosystem II), while females demonstrated higher values and better recovery after desiccation. Protonemata also showed variation in Fv/Fm over time and with sex, with no significant differences in ϕPSII between them. Desiccated male shoots had higher mortality rates and produced fewer new shoots. For females, the regeneration patterns varied between the desiccation-exposed groups and the control, with decreased shoot production, and some protonemata growing into filaments without forming shoots. CONCLUSION: These findings improve our understanding of the ecological responses of bryophytes to desiccation stress and provide insights into their adaptive strategies in challenging environments, such as the possible rarity of males in dioicous moss populations.


Assuntos
Dessecação , Adaptação Fisiológica/fisiologia , Brotos de Planta/fisiologia , Brotos de Planta/crescimento & desenvolvimento , Bryopsida/fisiologia , Reprodução/fisiologia , Brasil
5.
G3 (Bethesda) ; 14(7)2024 Jul 08.
Artigo em Inglês | MEDLINE | ID: mdl-38781445

RESUMO

The first chromosome-scale reference genome of the rare narrow-endemic African moss Physcomitrellopsis africana (P. africana) is presented here. Assembled from 73 × Oxford Nanopore Technologies (ONT) long reads and 163 × Beijing Genomics Institute (BGI)-seq short reads, the 414 Mb reference comprises 26 chromosomes and 22,925 protein-coding genes [Benchmarking Universal Single-Copy Ortholog (BUSCO) scores: C:94.8% (D:13.9%)]. This genome holds 2 genes that withstood rigorous filtration of microbial contaminants, have no homolog in other land plants, and are thus interpreted as resulting from 2 unique horizontal gene transfers (HGTs) from microbes. Further, P. africana shares 176 of the 273 published HGT candidates identified in Physcomitrium patens (P. patens), but lacks 98 of these, highlighting that perhaps as many as 91 genes were acquired in P. patens in the last 40 million years following its divergence from its common ancestor with P. africana. These observations suggest rather continuous gene gains via HGT followed by potential losses during the diversification of the Funariaceae. Our findings showcase both dynamic flux in plant HGTs over evolutionarily "short" timescales, alongside enduring impacts of successful integrations, like those still functionally maintained in extant P. africana. Furthermore, this study describes the informatic processes employed to distinguish contaminants from candidate HGT events.


Assuntos
Bryopsida , Transferência Genética Horizontal , Genoma de Planta , Filogenia , Bryopsida/genética , Genômica/métodos , Anotação de Sequência Molecular
6.
Front Immunol ; 15: 1383123, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38799460

RESUMO

Most drugs that target the complement system are designed to inhibit the complement pathway at either the proximal or terminal levels. The use of a natural complement regulator such as factor H (FH) could provide a superior treatment option by restoring the balance of an overactive complement system while preserving its normal physiological functions. Until now, the systemic treatment of complement-associated disorders with FH has been deemed unfeasible, primarily due to high production costs, risks related to FH purified from donors' blood, and the challenging expression of recombinant FH in different host systems. We recently demonstrated that a moss-based expression system can produce high yields of properly folded, fully functional, recombinant FH. However, the half-life of the initial variant (CPV-101) was relatively short. Here we show that the same polypeptide with modified glycosylation (CPV-104) achieves a pharmacokinetic profile comparable to that of native FH derived from human serum. The treatment of FH-deficient mice with CPV-104 significantly improved important efficacy parameters such as the normalization of serum C3 levels and the rapid degradation of C3 deposits in the kidney compared to treatment with CPV-101. Furthermore, CPV-104 showed comparable functionality to serum-derived FH in vitro, as well as similar performance in ex vivo assays involving samples from patients with atypical hemolytic uremic syndrome, C3 glomerulopathy and paroxysomal nocturnal hematuria. CPV-104 - the human FH analog expressed in moss - will therefore allow the treatment of complement-associated human diseases by rebalancing instead of inhibiting the complement cascade.


Assuntos
Fator H do Complemento , Humanos , Fator H do Complemento/metabolismo , Fator H do Complemento/genética , Animais , Camundongos , Meia-Vida , Polissacarídeos/metabolismo , Bryopsida/metabolismo , Bryopsida/genética , Glicosilação , Proteínas Recombinantes , Camundongos Knockout , Camundongos Endogâmicos C57BL , Masculino
7.
J Plant Physiol ; 297: 154253, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38703549

RESUMO

Moss plants appear in the early stages of land colonization and possess varying degrees of dehydration tolerance. In this study, a protein called PpFAS1.3 was identified, which contains a fasciclin 1-like domain and is essential for the moss Physcomitrium patens' response to short-term rapid dehydration. When the FAS1.3 protein was knocked out, leafyshoots showed a significant decrease in tolerance to rapid dehydration, resulting in accelerated water loss and increased membrane leakage. Phylogenetic analysis suggests that PpFAS1.3 and its homologous proteins may have originated from bacteria and are specifically found in non-vascular plants like mosses and liverworts. As a dehydration-related protein, FAS1.3 plays a significant role in regulating lipid metabolism, particularly in the synthesis of free fatty acids (FFA) and the metabolism of two phospholipids, PC and PA. This discovery highlights the close connection between PpFAS1.3 and lipid metabolism, providing new insights into the molecular mechanisms underlying plant adaptation to stresses.


Assuntos
Bryopsida , Metabolismo dos Lipídeos , Filogenia , Proteínas de Plantas , Proteínas de Plantas/metabolismo , Proteínas de Plantas/genética , Bryopsida/metabolismo , Bryopsida/genética , Desidratação , Regulação da Expressão Gênica de Plantas , Sequência de Aminoácidos
8.
Sci Total Environ ; 932: 173045, 2024 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-38734098

RESUMO

The main objective of this study was to develop and test a method of separating externally deposited Mn oxyhydroxides and co-precipitated elements from samples of aquatic moss (the moss Fontinalis antipyretica). The method, which uses 0.1 M hydroxylamine to dissolve the oxyhydroxides, was tested with samples collected in rivers with slightly acidic, well­oxygenated waters, where high rates of Mn precipitation occur. The method was effective (it extracted up to 84 % of the Mn) and selective (Fe oxyhydroxides were not extracted). The elements Ba, Cd, Zn and Ni were associated with the Mn oxyhydroxides, while Al, As, Cr, Cu, Fe, Hg and Pb were not. Deposition of Mn therefore increased the concentration of some elements in the moss samples. However, as Mn precipitation depends on Eh and pH, which are independent of the concentrations of the elements in water, the relationship between water and moss element concentrations is not clear (i.e. the data are noisy). This is a problem in biomonitoring studies, which assume a close relationship between element concentrations in moss and water. The value of the proposed extraction method is that it can be used to correct the effect of Mn deposition. We present an example of this correction applied to the Cd concentrations in the test data. We found that the noise introduced by the Mn, including age-related effects (observed by comparing concentrations in 0-2.5 and 2.2-5.0 cm sections from the shoot apex), can be reduced. Additionally, the correction revealed recent increases in Cd concentrations in one site that were not observed in the uncorrected data. Another finding of interest was the low content of total Mn and different extractability (of most elements) observed in moss samples collected in alkaline waters. Finally, we discuss how future studies designed for different environmental scenarios can benefit from application of the proposed method.


Assuntos
Briófitas , Monitoramento Ambiental , Oligoelementos , Poluentes Químicos da Água , Oligoelementos/análise , Poluentes Químicos da Água/análise , Monitoramento Ambiental/métodos , Briófitas/química , Manganês/análise , Bryopsida/química
9.
Plant Physiol Biochem ; 212: 108777, 2024 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-38820915

RESUMO

This study investigated the responses of Didymodon constrictus and Hypnum plumaeforme to different light qualities emitted by light-emitting diodes (LEDs), including white light (WL), red light (RL), blue light (BL), yellow light (YL), green light (GL), and a combination of red and blue light (R1B1L). The research analyzed the fluorescence imaging, photosynthetic pigments, coloration, and growth characteristics related to antioxidant enzymes in these two moss species. The results indicated that R1B1L significantly enhanced the content of photosynthetic pigments, maximum relative electron transport rate (rETRmax), saturation light intensity (IK), and the greenness of the moss. RL improved the maximum quantum yield (Fv/Fm), the light energy efficiency of H. plumaeforme and effective quantum yield in both moss species. In contrast, BL notably increased non-photochemical quenching (NPQ), photochemical quenching (qp), and the steady-state fluorescence decrease ratio (RFD) in H. plumaeforme. The application of GL significantly increases the maximum photon yield (Fv/Fm) in D. constrictus, as well as the light energy efficiency and elongation length, resulting in a shift in the color composition of both moss species towards yellow. Among the light treatments, R1B1L had the highest induction rate and promotional effect on the growth of both moss species. These mosses absorbed GL and RL effectively, while BL played a crucial role in the dissipation of heat and electron transfer in H. plumaeforme. This research provides valuable insights for the regulation of LED light environments and the physiological adaptability of moss in artificial cultivation.


Assuntos
Clorofila , Luz , Clorofila/metabolismo , Fluorescência , Briófitas/metabolismo , Briófitas/efeitos da radiação , Briófitas/crescimento & desenvolvimento , Fotossíntese/efeitos da radiação , Fotossíntese/fisiologia , Bryopsida/metabolismo , Bryopsida/efeitos da radiação , Bryopsida/crescimento & desenvolvimento
10.
Plant J ; 119(2): 1091-1111, 2024 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-38642374

RESUMO

Green feather algae (Bryopsidales) undergo a unique life cycle in which a single cell repeatedly executes nuclear division without cytokinesis, resulting in the development of a thallus (>100 mm) with characteristic morphology called coenocyte. Bryopsis is a representative coenocytic alga that has exceptionally high regeneration ability: extruded cytoplasm aggregates rapidly in seawater, leading to the formation of protoplasts. However, the genetic basis of the unique cell biology of Bryopsis remains poorly understood. Here, we present a high-quality assembly and annotation of the nuclear genome of Bryopsis sp. (90.7 Mbp, 27 contigs, N50 = 6.7 Mbp, 14 034 protein-coding genes). Comparative genomic analyses indicate that the genes encoding BPL-1/Bryohealin, the aggregation-promoting lectin, are heavily duplicated in Bryopsis, whereas homologous genes are absent in other ulvophyceans, suggesting the basis of regeneration capability of Bryopsis. Bryopsis sp. possesses >30 kinesins but only a single myosin, which differs from other green algae that have multiple types of myosin genes. Consistent with this biased motor toolkit, we observed that the bidirectional motility of chloroplasts in the cytoplasm was dependent on microtubules but not actin in Bryopsis sp. Most genes required for cytokinesis in plants are present in Bryopsis, including those in the SNARE or kinesin superfamily. Nevertheless, a kinesin crucial for cytokinesis initiation in plants (NACK/Kinesin-7II) is hardly expressed in the coenocytic part of the thallus, possibly underlying the lack of cytokinesis in this portion. The present genome sequence lays the foundation for experimental biology in coenocytic macroalgae.


Assuntos
Genoma de Planta , Genoma de Planta/genética , Filogenia , Clorófitas/genética , Clorófitas/fisiologia , Regeneração/genética , Bryopsida/genética , Bryopsida/fisiologia , Bryopsida/citologia , Cinesinas/genética , Cinesinas/metabolismo , Miosinas/genética , Miosinas/metabolismo
11.
Transgenic Res ; 33(1-2): 67-74, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38573428

RESUMO

Genome editing via CRISPR/Cas has enabled targeted genetic modifications in various species, including plants. The requirement for specific protospacer-adjacent motifs (PAMs) near the target gene, as seen with Cas nucleases like SpCas9, limits its application. PAMless SpCas9 variants, designed with a relaxed PAM requirement, have widened targeting options. However, these so-call PAMless SpCas9 still show variation of editing efficiency depending on the PAM and their efficiency lags behind the native SpCas9. Here we assess the potential of a PAMless SpCas9 variant for genome editing in the model plant Physcomitrium patens. For this purpose, we developed a SpRYCas9i variant, where expression was optimized, and tested its editing efficiency using the APT as a reporter gene. We show that the near PAMless SpRYCas9i effectively recognizes specific PAMs in P. patens that are not or poorly recognized by the native SpCas9. Pattern of mutations found using the SpRYCas9i are similar to the ones found with the SpCas9 and we could not detect off-target activity for the sgRNAs tested in this study. These findings contribute to advancing versatile genome editing techniques in plants.


Assuntos
Bryopsida , Edição de Genes , Edição de Genes/métodos , Sistemas CRISPR-Cas/genética , Proteína 9 Associada à CRISPR/genética , Proteína 9 Associada à CRISPR/metabolismo , RNA Guia de Sistemas CRISPR-Cas , Mutação , Bryopsida/genética , Genoma de Planta/genética
12.
New Phytol ; 242(5): 1996-2010, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38571393

RESUMO

The conquest of land by plants was concomitant with, and possibly enabled by, the evolution of three-dimensional (3D) growth. The moss Physcomitrium patens provides a model system for elucidating molecular mechanisms in the initiation of 3D growth. Here, we investigate whether the phytohormone ethylene, which is believed to have been a signal before land plant emergence, plays a role in 3D growth regulation in P. patens. We report ethylene controls 3D gametophore formation, based on results from exogenously applied ethylene and genetic manipulation of PpEIN2, which is a central component in the ethylene signaling pathway. Overexpression (OE) of PpEIN2 activates ethylene responses and leads to earlier formation of gametophores with fewer gametophores produced thereafter, phenocopying ethylene-treated wild-type. Conversely, Ppein2 knockout mutants, which are ethylene insensitive, show initially delayed gametophore formation with more gametophores produced later. Furthermore, pharmacological and biochemical analyses reveal auxin levels are decreased in the OE lines but increased in the knockout mutants. Our results suggest that evolutionarily, ethylene and auxin molecular networks were recruited to build the plant body plan in ancestral land plants. This might have played a role in enabling ancient plants to acclimate to the continental surfaces of the planet.


Assuntos
Bryopsida , Etilenos , Regulação da Expressão Gênica de Plantas , Ácidos Indolacéticos , Proteínas de Plantas , Etilenos/metabolismo , Ácidos Indolacéticos/metabolismo , Ácidos Indolacéticos/farmacologia , Bryopsida/crescimento & desenvolvimento , Bryopsida/genética , Bryopsida/efeitos dos fármacos , Bryopsida/metabolismo , Proteínas de Plantas/metabolismo , Proteínas de Plantas/genética , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Células Germinativas Vegetais/metabolismo , Células Germinativas Vegetais/crescimento & desenvolvimento , Células Germinativas Vegetais/efeitos dos fármacos , Mutação/genética
13.
Biochem Soc Trans ; 52(2): 505-515, 2024 Apr 24.
Artigo em Inglês | MEDLINE | ID: mdl-38629612

RESUMO

In eukaryotic cells, organelle and vesicle transport, positioning, and interactions play crucial roles in cytoplasmic organization and function. These processes are governed by intracellular trafficking mechanisms. At the core of that trafficking, the cytoskeleton and directional transport by motor proteins stand out as its key regulators. Plant cell tip growth is a well-studied example of cytoplasm organization by polarization. This polarization, essential for the cell's function, is driven by the cytoskeleton and its associated motors. This review will focus on myosin XI, a molecular motor critical for vesicle trafficking and polarized plant cell growth. We will center our discussion on recent data from the moss Physcomitrium patens and the liverwort Marchantia polymorpha. The biochemical properties and structure of myosin XI in various plant species are discussed, highlighting functional conservation across species. We further explore this conservation of myosin XI function in the process of vesicle transport in tip-growing cells. Existing evidence indicates that myosin XI actively organizes actin filaments in tip-growing cells by a mechanism based on vesicle clustering at their tips. A hypothetical model is presented to explain the essential function of myosin XI in polarized plant cell growth based on vesicle clustering at the tip. The review also provides insight into the in vivo localization and dynamics of myosin XI, emphasizing its role in cytosolic calcium regulation, which influences the polymerization of F-actin. Lastly, we touch upon the need for additional research to elucidate the regulation of myosin function.


Assuntos
Miosinas , Células Vegetais , Miosinas/metabolismo , Células Vegetais/metabolismo , Bryopsida/metabolismo , Bryopsida/crescimento & desenvolvimento , Proteínas de Plantas/metabolismo , Citoesqueleto de Actina/metabolismo , Marchantia/metabolismo , Marchantia/crescimento & desenvolvimento , Desenvolvimento Vegetal/fisiologia
14.
New Phytol ; 242(5): 2251-2269, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38501480

RESUMO

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.


Assuntos
Ceras , Ceras/metabolismo , Álcoois/metabolismo , Filogenia , Marchantia/genética , Marchantia/metabolismo , Proteínas de Plantas/metabolismo , Proteínas de Plantas/genética , Bryopsida/genética , Bryopsida/metabolismo , Briófitas/genética , Briófitas/metabolismo , Aldeído Oxirredutases/metabolismo , Aldeído Oxirredutases/genética , Vias Biossintéticas/genética , Evolução Molecular , Regulação da Expressão Gênica de Plantas , Aciltransferases/metabolismo , Aciltransferases/genética , Evolução Biológica , Arabidopsis/genética , Arabidopsis/metabolismo , Mutação/genética
15.
Planta ; 259(5): 92, 2024 Mar 20.
Artigo em Inglês | MEDLINE | ID: mdl-38504021

RESUMO

MAIN CONCLUSION: Fiber-like cells with thickened cell walls of specific structure and polymer composition that includes (1 → 4)-ß-galactans develop in the outer stem cortex of several moss species gametophytes. The early land plants evolved several specialized cell types and tissues that did not exist in their aquatic ancestors. Of these, water-conducting elements and reproductive organs have received most of the research attention. The evolution of tissues specialized to fulfill a mechanical function is by far less studied despite their wide distribution in land plants. For vascular plants following a homoiohydric trajectory, the evolutionary emergence of mechanical tissues is mainly discussed starting with the fern-like plants with their hypodermal sterome or sclerified fibers that have xylan and lignin-based cell walls. However, mechanical challenges were also faced by bryophytes, which lack lignified cell-walls. To characterize mechanical tissues in the bryophyte lineage, following a poikilohydric trajectory, we used six wild moss species (Polytrichum juniperinum, Dicranum sp., Rhodobryum roseum, Eurhynchiadelphus sp., Climacium dendroides, and Hylocomium splendens) and analyzed the structure and composition of their cell walls. In all of them, the outer stem cortex of the leafy gametophytic generation had fiber-like cells with a thickened but non-lignified cell wall. Such cells have a spindle-like shape with pointed tips. The additional thick cell wall layer in those fiber-like cells is composed of sublayers with structural evidence for different cellulose microfibril orientation, and with specific polymer composition that includes (1 → 4)-ß-galactans. Thus, the basic cellular characters of the cells that provide mechanical support in vascular plant taxa (elongated cell shape, location at the periphery of a primary organ, the thickened cell wall and its peculiar composition and structure) also exist in mosses.


Assuntos
Briófitas , Bryopsida , Células Germinativas Vegetais/metabolismo , Plantas/metabolismo , Bryopsida/metabolismo , Lignina/metabolismo , Galactanos/metabolismo , Parede Celular/metabolismo
16.
Methods Mol Biol ; 2758: 375-385, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38549025

RESUMO

Here, we report our approach to peptidomic analysis of the plant model Physcomitrium patens. Intracellular and extracellular peptides were extracted under conditions preventing proteolytic digestion by endogenous proteases. The extracts were fractionated on size exclusion columns to isolate intracellular peptides and on reversed-phase cartridges to isolate extracellular peptides, with the isolated peptides subjected to LC-MS/MS analysis. Mass spectrometry data were analyzed for the presence of peptides derived from the known proteins or microproteins encoded by small open reading frames (<100 aa, smORFs) predicted in the moss genome. Experimental details are provided for each step.


Assuntos
Bryopsida , Espectrometria de Massas em Tandem , Cromatografia Líquida/métodos , Espectrometria de Massas em Tandem/métodos , Peptídeos/química , Proteínas/metabolismo , Peptídeo Hidrolases/metabolismo , Bryopsida/metabolismo
17.
Microb Ecol ; 87(1): 49, 2024 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-38427046

RESUMO

Moss-cyanobacteria symbioses were proposed to be based on nutrient exchange, with hosts providing C and S while bacteria provide N, but we still lack understanding of the underlying molecular mechanisms of their interactions. We investigated how contact between the ubiquitous moss Hylocomium splendens and its cyanobiont affects nutrient-related gene expression of both partners. We isolated a cyanobacterium from H. splendens and co-incubated it with washed H. splendens shoots. Cyanobacterium and moss were also incubated separately. After 1 week, we performed acetylene reduction assays to estimate N2 fixation and RNAseq to evaluate metatranscriptomes. Genes related to N2 fixation and the biosynthesis of several amino acids were up-regulated in the cyanobiont when hosted by the moss. However, S-uptake and the biosynthesis of the S-containing amino acids methionine and cysteine were down-regulated in the cyanobiont while the degradation of selenocysteine was up-regulated. In contrast, the number of differentially expressed genes in the moss was much lower, and almost no transcripts related to nutrient metabolism were affected. It is possible that, at least during the early stage of this symbiosis, the cyanobiont receives few if any nutrients from the host in return for N, suggesting that moss-cyanobacteria symbioses encompass relationships that are more plastic than a constant mutualist flow of nutrients.


Assuntos
Briófitas , Bryopsida , Cianobactérias , Simbiose , Fixação de Nitrogênio , Bryopsida/genética , Bryopsida/metabolismo , Bryopsida/microbiologia , Cianobactérias/metabolismo , Aminoácidos/metabolismo
18.
Adv Sci (Weinh) ; 11(20): e2306767, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38552153

RESUMO

Plant movements for survival are nontrivial. Antheridia in the moss Physcomitrium patens (P. patens) use motion to eject sperm in the presence of water. However, the biological and mechanical mechanisms that actuate the process are unknown. Here, the burst of the antheridium of P. patens, triggered by water, results from elastic instability and is determined by an asymmetric change in cell geometry. The tension generated in jacket cell walls of antheridium arises from turgor pressure, and is further promoted when the inner walls of apex burst in hydration, causing water and cellular contents of apex quickly influx into sperm chamber. The outer walls of the jacket cells are strengthened by NAC transcription factor VNS4 and serve as key morphomechanical innovations to store hydrostatic energy in a confined space in P. patens. However, the antheridium in liverwort Marchantia polymorpha (M. polymorpha) adopts a different strategy for sperm release; like jacket cell outer walls of P. patens, the cells surrounding the antheridium of M. polymorpha appear to play a similar role in the storage of energy. Collectively, the work shows that plants have evolved different ingenious devices for sperm discharge and that morphological innovations can differ.


Assuntos
Bryopsida , Bryopsida/fisiologia , Bryopsida/citologia , Bryopsida/metabolismo , Marchantia/genética , Marchantia/metabolismo , Marchantia/citologia , Marchantia/fisiologia , Briófitas/fisiologia , Briófitas/metabolismo
19.
Commun Biol ; 7(1): 261, 2024 Mar 04.
Artigo em Inglês | MEDLINE | ID: mdl-38438476

RESUMO

Calpains are cysteine proteases that control cell fate transitions whose loss of function causes severe, pleiotropic phenotypes in eukaryotes. Although mainly considered as modulatory proteases, human calpain targets are directed to the N-end rule degradation pathway. Several such targets are transcription factors, hinting at a gene-regulatory role. Here, we analyze the gene-regulatory networks of the moss Physcomitrium patens and characterize the regulons that are misregulated in mutants of the calpain DEFECTIVE KERNEL1 (DEK1). Predicted cleavage patterns of the regulatory hierarchies in five DEK1-controlled subnetworks are consistent with a pleiotropic and regulatory role during cell fate transitions targeting multiple functions. Network structure suggests DEK1-gated sequential transitions between cell fates in 2D-to-3D development. Our method combines comprehensive phenotyping, transcriptomics and data science to dissect phenotypic traits, and our model explains the protease function as a switch gatekeeping cell fate transitions potentially also beyond plant development.


Assuntos
Bryopsida , Peptídeo Hidrolases , Humanos , Calpaína/genética , Endopeptidases , Diferenciação Celular/genética
20.
New Phytol ; 243(3): 981-996, 2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-38415863

RESUMO

Water scarcity, resulting from climate change, poses a significant threat to ecosystems. Syntrichia ruralis, a dryland desiccation-tolerant moss, provides valuable insights into survival of water-limited conditions. We sequenced the genome of S. ruralis, conducted transcriptomic analyses, and performed comparative genomic and transcriptomic analyses with existing genomes and transcriptomes, including with the close relative S. caninervis. We took a genetic approach to characterize the role of an S. ruralis transcription factor, identified in transcriptomic analyses, in Arabidopsis thaliana. The genome was assembled into 12 chromosomes encompassing 21 169 protein-coding genes. Comparative analysis revealed copy number and transcript abundance differences in known desiccation-associated gene families, and highlighted genome-level variation among species that may reflect adaptation to different habitats. A significant number of abscisic acid (ABA)-responsive genes were found to be negatively regulated by a MYB transcription factor (MYB55) that was upstream of the S. ruralis ortholog of ABA-insensitive 3 (ABI3). We determined that this conserved MYB transcription factor, uncharacterized in Arabidopsis, acts as a negative regulator of an ABA-dependent stress response in Arabidopsis. The new genomic resources from this emerging model moss offer novel insights into how plants regulate their responses to water deprivation.


Assuntos
Arabidopsis , Dessecação , Regulação da Expressão Gênica de Plantas , Genoma de Planta , Arabidopsis/genética , Arabidopsis/fisiologia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Ácido Abscísico/farmacologia , Ácido Abscísico/metabolismo , Filogenia , Sequência Conservada/genética , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Bryopsida/genética , Bryopsida/fisiologia , Genes de Plantas , Estresse Fisiológico/genética , Modelos Biológicos , Transcriptoma/genética
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