Crossroads in the evolution of plant specialized metabolism.

The monophyletic group of embryophytes (land plants) stands out among photosynthetic eukaryotes: they are the sole constituents of the macroscopic flora on land. In their entirety, embryophytes account for the majority of the biomass on land and constitute an astounding biodiversity. What allowed for the massive radiation of this particular lineage? One of the defining features of all land plants is the production of an array of specialized metabolites. The compounds that the specialized metabolic pathways of embryophytes produce have diverse functions, ranging from superabundant structural polymers and compounds that ward off abiotic and biotic challenges, to signaling molecules whose abundance is measured at the nanomolar scale. These specialized metabolites govern the growth, development, and physiology of land plants-including their response to the environment. Hence, specialized metabolites define the biology of land plants as we know it. And they were likely a foundation for their success. It is thus intriguing to find that the closest algal relatives of land plants, freshwater organisms from the grade of streptophyte algae, possess homologs for key enzymes of specialized metabolic pathways known from land plants. Indeed, some studies suggest that signature metabolites emerging from these pathways can be found in streptophyte algae. Here we synthesize the current understanding of which routes of the specialized metabolism of embryophytes can be traced to a time before plants had conquered land.

Evolutionary and immune-activating character analyses of NLR genes in algae suggest the ancient origin of plant intracellular immune receptors.

Nucleotide-binding leucine-rich repeat (NLR) proteins are crucial intracellular immune receptors in plants, responsible for detecting invading pathogens and initiating defense responses. While previous studies on the evolution and function of NLR genes were mainly limited to land plants, the evolutionary trajectory and immune-activating character of NLR genes in algae remain less explored. In this study, genome-wide NLR gene analysis was conducted on 44 chlorophyte species across seven classes and seven charophyte species across five classes. A few but variable number of NLR genes, ranging from one to 20, were identified in five chlorophytes and three charophytes, whereas no NLR gene was identified from the remaining algal genomes. Compared with land plants, algal genomes possess fewer or usually no NLR genes, implying that the expansion of NLR genes in land plants can be attributed to their adaptation to the more complex terrestrial pathogen environments. Through phylogenetic analysis, domain composition analysis, and conserved motifs profiling of the NBS domain, we detected shared and lineage-specific features between NLR genes in algae and land plants, supporting the common origin and continuous evolution of green plant NLR genes. Immune-activation assays revealed that both TNL and RNL proteins from green algae can elicit hypersensitive responses in Nicotiana benthamiana, indicating the molecular basis for immune activation has emerged in the early evolutionary stage of different types of NLR proteins. In summary, the results from this study suggest that NLR proteins may have taken a role as intracellular immune receptors in the common ancestor of green plants.

Endosidin 5 disruption of the Golgi apparatus and extracellular matrix secretion in the unicellular charophyte Penium margaritaceum.

BACKGROUND AND AIMS: Endosidins are a group of low-molecular-weight compounds, first identified by 'chemical biology' screening assays, that have been used to target specific components of the endomembrane system. In this study, we employed multiple microscopy-based screening techniques to elucidate the effects of endosidin 5 (ES5) on the Golgi apparatus and the secretion of extracellular matrix (ECM) components in Penium margaritaceum. These effects were compared with those caused by treatments with brefeldin A and concanamycin A. Penium margaritaceum's extensive Golgi apparatus and endomembrane system make it an outstanding model organism for screening changes to the endomembrane system. Here we detail changes to the Golgi apparatus and secretion of ECM material caused by ES5. METHODS: Changes to extracellular polymeric substance (EPS) secretion and cell wall expansion were screened using fluorescence microscopy. Confocal laser scanning microscopy and transmission electron microscopy were used to assess changes to the Golgi apparatus, the cell wall and the vesicular network. Electron tomography was also performed to detail the changes to the Golgi apparatus. KEY RESULTS: While other endosidins were able to impact EPS secretion and cell wall expansion, only ES5 completely inhibited EPS secretion and cell wall expansion over 24 h. Short treatments of ES5 resulted in displacement of the Golgi bodies from their typical linear alignment. The number of cisternae decreased per Golgi stack and trans face cisternae in-curled to form distinct elongate circular profiles. Longer treatment resulted in a transformation of the Golgi body to an irregular aggregate of cisternae. These alterations could be reversed by removal of ES5 and returning cells to culture. CONCLUSIONS: ES5 alters secretion of ECM material in Penium by affecting the Golgi apparatus and does so in a markedly different way from other endomembrane inhibitors such as brefeldin A and concanamycin A.

Green land: Multiple perspectives on green algal evolution and the earliest land plants.

Green plants, broadly defined as green algae and the land plants (together, Viridiplantae), constitute the primary eukaryotic lineage that successfully colonized Earth's emergent landscape. Members of various clades of green plants have independently made the transition from fully aquatic to subaerial habitats many times throughout Earth's history. The transition, from unicells or simple filaments to complex multicellular plant bodies with functionally differentiated tissues and organs, was accompanied by innovations built upon a genetic and phenotypic toolkit that have served aquatic green phototrophs successfully for at least a billion years. These innovations opened an enormous array of new, drier places to live on the planet and resulted in a huge diversity of land plants that have dominated terrestrial ecosystems over the past 500 million years. This review examines the greening of the land from several perspectives, from paleontology to phylogenomics, to water stress responses and the genetic toolkit shared by green algae and plants, to the genomic evolution of the sporophyte generation. We summarize advances on disparate fronts in elucidating this important event in the evolution of the biosphere and the lacunae in our understanding of it. We present the process not as a step-by-step advancement from primitive green cells to an inevitable success of embryophytes, but rather as a process of adaptations and exaptations that allowed multiple clades of green plants, with various combinations of morphological and physiological terrestrialized traits, to become diverse and successful inhabitants of the land habitats of Earth.

Heat stress response in the closest algal relatives of land plants reveals conserved stress signaling circuits.

All land plants (embryophytes) share a common ancestor that likely evolved from a filamentous freshwater alga. Elucidating the transition from algae to embryophytes - and the eventual conquering of Earth's surface - is one of the most fundamental questions in plant evolutionary biology. Here, we investigated one of the organismal properties that might have enabled this transition: resistance to drastic temperature shifts. We explored the effect of heat stress in Mougeotia and Spirogyra, two representatives of Zygnematophyceae - the closest known algal sister lineage to land plants. Heat stress induced pronounced phenotypic alterations in their plastids, and high-performance liquid chromatography-tandem mass spectroscopy-based profiling of 565 transitions for the analysis of main central metabolites revealed significant shifts in 43 compounds. We also analyzed the global differential gene expression responses triggered by heat, generating 92.8 Gbp of sequence data and assembling a combined set of 8905 well-expressed genes. Each organism had its own distinct gene expression profile; less than one-half of their shared genes showed concordant gene expression trends. We nevertheless detected common signature responses to heat such as elevated transcript levels for molecular chaperones, thylakoid components, and - corroborating our metabolomic data - amino acid metabolism. We also uncovered the heat-stress responsiveness of genes for phosphorelay-based signal transduction that links environmental cues, calcium signatures and plastid biology. Our data allow us to infer the molecular heat stress response that the earliest land plants might have used when facing the rapidly shifting temperature conditions of the terrestrial habitat.

Evo-physio: on stress responses and the earliest land plants.

Embryophytes (land plants) can be found in almost any habitat on the Earth's surface. All of this ecologically diverse embryophytic flora arose from algae through a singular evolutionary event. Traits that were, by their nature, indispensable for the singular conquest of land by plants were those that are key for overcoming terrestrial stressors. Not surprisingly, the biology of land plant cells is shaped by a core signaling network that connects environmental cues, such as stressors, to the appropriate responses-which, thus, modulate growth and physiology. When did this network emerge? Was it already present when plant terrestrialization was in its infancy? A comparative approach between land plants and their algal relatives, the streptophyte algae, allows us to tackle such questions and resolve parts of the biology of the earliest land plants. Exploring the biology of the earliest land plants might shed light on exactly how they overcame the challenges of terrestrialization. Here, we outline the approaches and rationale underlying comparative analyses towards inferring the genetic toolkit for the stress response that aided the earliest land plants in their conquest of land.

Shallow plant-dominated lakes - extreme environmental variability, carbon cycling and ecological species challenges.

BACKGROUND: Submerged plants composed of charophytes (green algae) and angiosperms develop dense vegetation in small, shallow lakes and in littoral zones of large lakes. Many small, oligotrophic plant species have declined due to drainage and fertilization of lakes, while some tall, eutrophic species have increased. Although plant distribution has been thoroughly studied, the physiochemical dynamics and biological challenges in plant-dominated lakes have been grossly understudied, even though they may offer the key to species persistence. SCOPE: Small plant-dominated lakes function as natural field laboratories with eco-physiological processes in dense vegetation dictating extreme environmental variability, intensive photosynthesis and carbon cycling. Those processes can be quantified on a whole lake basis at high temporal resolution by continuously operating sensors for light, temperature, oxygen, etc. We explore this hitherto hidden world. CONCLUSIONS: Dense plant canopies attenuate light and wind-driven turbulence and generate separation between warm surface water and colder bottom waters. Daytime vertical stratification becomes particularly strong in dense charophyte vegetation, but stratification is a common feature in small, shallow lakes also without plants. Surface cooling at night induces mixing of the water column. Daytime stratification in plant stands may induce hypoxia or anoxia in dark bottom waters by respiration, while surface waters develop oxygen supersaturation by photosynthesis. Intensive photosynthesis and calcification in shallow charophyte lakes depletes dissolved inorganic carbon (DIC) in surface waters, whereas DIC is replenished by respiration and carbonate dissolution in bottom waters and returned to surface waters before sunrise. Extreme diel changes in temperature, DIC and oxygen in dense vegetation can induce extensive rhythmicity of photosynthesis and respiration and become a severe challenge to the survival of organisms. Large phosphorus pools are bound in plant tissue and carbonate precipitates. Future studies should test the importance of this phosphorus sink for ecosystem processes and competition between phytoplankton and plants.

Plant evolution: landmarks on the path to terrestrial life.

Contents Summary 1428 I. The singularity of plant terrestrialization 1428 II. Adaptation vs exaptation - what shaped the land plant toolkit? 1430 III. Trait mosaicism in (higher-branching) streptophyte algae 1431 IV. CONCLUSIONS: a streptophyte algal perspective on land plant trait evolution 1432 Acknowledgements 1432 ORCID 1433 References 1433 SUMMARY: Photosynthetic eukaryotes thrive anywhere there is sunlight and water. But while such organisms are exceptionally diverse in form and function, only one phototrophic lineage succeeded in rising above its substrate: the land plants (embryophytes). Molecular phylogenetic data show that land plants evolved from streptophyte algae most closely related to extant Zygnematophyceae, and one of the principal aims of plant evolutionary biology is to uncover the key features of such algae that enabled this important transition. At the present time, however, mosaic and reductive evolution blur our picture of the closest algal ancestors of plants. Here we discuss recent progress and problems in inferring the biology of the algal progenitor of the terrestrial photosynthetic macrobiome.

Evolution of nuclear auxin signaling: lessons from genetic studies with basal land plants.

Auxin plays critical roles in growth and development through the regulation of cell differentiation, cell expansion, and pattern formation. The auxin signal is mainly conveyed through a so-called nuclear auxin pathway involving the receptor TIR1/AFB, the transcriptional co-repressor AUX/IAA, and the transcription factor ARF with direct DNA-binding ability. Recent progress in sequence information and molecular genetics in basal plants has provided many insights into the evolutionary origin of the nuclear auxin pathway and its pleiotropic roles in land plant development. In this review, we summarize the latest knowledge of the nuclear auxin pathway gained from studies using basal plants, including charophycean green algae and two major model bryophytes, Marchantia polymorpha and Physcomitrella patens. In addition, we discuss the functional implication of the increase in genetic complexity of the nuclear auxin pathway during land plant evolution.

How Embryophytic is the Biosynthesis of Phenylpropanoids and their Derivatives in Streptophyte Algae?

The origin of land plants from algae is a long-standing question in evolutionary biology. It is becoming increasingly clear that many characters that were once assumed to be 'embryophyte specific' can in fact be found in their closest algal relatives, the streptophyte algae. One such case is the phenylpropanoid pathway. While biochemical data indicate that streptophyte algae harbor lignin-like components, the phenylpropanoid core pathway, which serves as the backbone of lignin biosynthesis, has been proposed to have arisen at the base of the land plants. Here we revisit this hypothesis using a wealth of new sequence data from streptophyte algae. Tracing the biochemical pathway towards lignin biogenesis, we show that most of the genes required for phenylpropanoid synthesis and the precursors for lignin production were already present in streptophyte algae. Nevertheless, phylogenetic analyses and protein structure predictions of one of the key enzyme classes in lignin production, cinnamyl alcohol dehydrogenase (CAD), suggest that CADs of streptophyte algae are more similar to sinapyl alcohol dehydrogenases (SADs). This suggests that the end-products of the pathway leading to lignin biosynthesis in streptophyte algae may facilitate the production of lignin-like compounds and defense molecules. We hypothesize that streptophyte algae already possessed the genetic toolkit from which the capacity to produce lignin later evolved in vascular plants.

Extreme diel dissolved oxygen and carbon cycles in shallow vegetated lakes.

A common perception in limnology is that shallow lakes are homogeneously mixed owing to their small water volume. However, this perception is largely gained by downscaling knowledge from large lakes to their smaller counterparts. Here we show that shallow vegetated lakes (less than 0.6 m), in fact, undergo recurring daytime stratification and nocturnal mixing accompanied by extreme chemical variations during summer. Dense submerged vegetation effectively attenuates light and turbulence generating separation between warm surface waters and much colder bottom waters. Photosynthesis in surface waters produces oxygen accumulation and CO2 depletion, whereas respiration in dark bottom waters causes anoxia and CO2 accumulation. High daytime pH in surface waters promotes precipitation of CaCO3 which is re-dissolved in bottom waters. Nocturnal convective mixing re-introduces oxygen into bottom waters for aerobic respiration and regenerated inorganic carbon into surface waters, which supports intense photosynthesis. Our results reconfigure the basic understanding of local environmental gradients in shallow lakes, one of the most abundant freshwater habitats globally.

Profound afternoon depression of ecosystem production and nighttime decline of respiration in a macrophyte-rich, shallow lake.

Small, shallow lakes with dense growth of submerged macrophytes are extremely abundant worldwide, but have remained grossly understudied although open water oxygen measurements should be suitable to determine diel fluctuations and test drivers of ecosystem metabolism during the day. We measured the temporal and spatial variability of environmental conditions as well as net ecosystem production (NEP) and respiration (R) in a small, shallow Swedish lake with dense charophyte stands by collecting data from oxygen-, pH-, temperature- and light-sensors across horizontal and vertical gradients during different periods between April and June in 3 years. We found reproducible diel oxygen patterns and daily metabolic rates. The charophyte canopy accounted for almost all primary production and respiration of the ecosystem. Two novel discoveries-profound afternoon depression of production and nighttime decline of respiration-occurred on virtually every day. Extensive increase of oxygen-, temperature- and pH-levels and depletion of dissolved inorganic carbon (DIC) and CO2 concentrations could account for maximum NEP-rates before noon and afternoon depression with low NEP-rates. Ecosystem respiration declined during the night to 24-70% of rates at sunset, probably because of depletion of respiratory substrates. Afternoon depression of photosynthesis should be widespread in numerous habitats with dense growth of macrophytes, periphyton, or phytoplankton implying that daily photosynthesis and growth are restricted and species with efficient DIC use may have an advantage.

Bioaccumulation and toxicity studies of macroalgae (Charophyceae) treated with aluminium: Experimental studies in the context of lake restoration.

The objective of this study was to examine the impact of aluminium on the perennial macroalgae Chara hispida L. and its bioaccumulation capacities. Aluminium (Al) was introduced into the environment in the form of polyaluminium chloride, an agent utilized in the restoration of waterbodies. Research was conducted in an experimental setting using mesocosms (volume 0.8m3) placed in the littoral zone of a lake with C. hispida. Three doses of the coagulant were applied, each with a different volume: low - 6.1g Al m-3, medium - 12.2gm-3 and high - 24.5g Al m-3. A significant acidification of environment was determined, which would imply the presence of toxic Al3+ ions. It has been demonstrated that aluminium penetrates and accumulates in the cells of the charophyte. This caused damage to the thalli, which manifested itself in chloroses, necroses, flaking of the cortex cells and softening of the thallus, whose severity was proportionate to the dose of the coagulant. The first negative signs were observed after 24h. The study shows that C. hispida is a poor accumulator of aluminium (bioconcentration factor < 200), while bioaccumulation capacity was inhibited at the concentration of approx. 2.0mg Al g-1 d.w. Accumulation in the thalli of the charophytes accounted for 58% of variation following removal of aluminium from the environment. The results of the experiment demonstrate a negative impact of aluminium on charophytes at concentrations used in aggressive restoration of lakes.

Sugar composition of the pectic polysaccharides of charophytes, the closest algal relatives of land-plants: presence of 3-O-methyl-D-galactose residues.

BACKGROUND AND AIMS: During evolution, plants have acquired and/or lost diverse sugar residues as cell-wall constituents. Of particular interest are primordial cell-wall features that existed, and in some cases abruptly changed, during the momentous step whereby land-plants arose from charophytic algal ancestors. METHODS: Polysaccharides were extracted from four charophyte orders [Chlorokybales (Chlorokybus atmophyticus), Klebsormidiales (Klebsormidium fluitans, K. subtile), Charales (Chara vulgaris, Nitella flexilis), Coleochaetales (Coleochaete scutata)] and an early-diverging land-plant (Anthoceros agrestis). 'Pectins' and 'hemicelluloses', operationally defined as extractable in oxalate (100 °C) and 6 m NaOH (37 °C), respectively, were acid- or Driselase-hydrolysed, and the monosaccharides analysed chromatographically. One unusual monosaccharide, 'U', was characterized by (1)H/(13)C-nuclear magnetic resonance spectroscopy and also enzymically. KEY RESULTS: 'U' was identified as 3-O-methyl-D-galactose (3-MeGal). All pectins, except in Klebsormidium, contained acid- and Driselase-releasable galacturonate, suggesting homogalacturonan. All pectins, without exception, released rhamnose and galactose on acid hydrolysis; however, only in 'higher' charophytes (Charales, Coleochaetales) and Anthoceros were these sugars also efficiently released by Driselase, suggesting rhamnogalacturonan-I. Pectins of 'higher' charophytes, especially Chara, contained little arabinose, instead possessing 3-MeGal. Anthoceros hemicelluloses were rich in glucose, xylose, galactose and arabinose (suggesting xyloglucan and arabinoxylan), none of which was consistently present in charophyte hemicelluloses. CONCLUSIONS: Homogalacturonan is an ancient streptophyte feature, albeit secondarily lost in Klebsormidium. When conquering the land, the first embryophytes already possessed rhamnogalacturonan-I. In contrast, charophyte and land-plant hemicelluloses differ substantially, indicating major changes during terrestrialization. The presence of 3-MeGal in charophytes and lycophytes but not in the 'intervening' bryophytes confirms that cell-wall chemistry changed drastically between major phylogenetic grades.

The capability to synthesize phytochelatins and the presence of constitutive and functional phytochelatin synthases are ancestral (plesiomorphic) characters for basal land plants.

Bryophytes, a paraphyletic group which includes liverworts, mosses, and hornworts, have been stated as land plants that under metal stress (particularly cadmium) do not synthesize metal-binding peptides such as phytochelatins. Moreover, very little information is available to date regarding phytochelatin synthesis in charophytes, postulated to be the direct ancestors of land plants, or in lycophytes, namely very basal tracheophytes. In this study, it was hypothesized that basal land plants and charophytes have the capability to produce phytochelatins and possess constitutive and functional phytochelatin synthases. To verify this hypothesis, twelve bryophyte species (six liverworts, four mosses, and two hornworts), three charophytes, and two lycophyte species were exposed to 0-36 µM cadmium for 72 h, and then assayed for: (i) glutathione and phytochelatin quali-quantitative content by HPLC and mass spectrometry; (ii) the presence of putative phytochelatin synthases by western blotting; and (iii) in vitro activity of phytochelatin synthases. Of all the species tested, ten produced phytochelatins in vivo, while the other seven did not. The presence of a constitutively expressed and functional phytochelatin synthase was demonstrated in all the bryophyte lineages and in the lycophyte Selaginella denticulata, but not in the charophytes. Hence, current knowledge according to phytochelatins have been stated as being absent in bryophytes was therefore confuted by this work. It is argued that the capability to synthesize phytochelatins, as well as the presence of active phytochelatin synthases, are ancestral (plesiomorphic) characters for basal land plants.

Rediscovering Chara as a model organism for molecular and evo-devo studies.

Chara has been used as a model for decades in the field of plant physiology, enabling the investigation of fundamental physiological processes. In electrophysiological studies, Chara has been utilized thanks to its large internodal cells that can be easily manipulated. Additionally, Chara played a pioneering role in elucidating the presence and function of the cytoskeleton in cytoplasmic streaming, predating similar findings in terrestrial plants. Its representation considerably declined following the establishment and routine application of genetic transformation techniques in Arabidopsis. Nevertheless, the recent surge in evo-devo studies can be attributed to the whole genome sequencing of the Chara braunii, which has shed light on ancestral traits prevalent in land plants. Surprisingly, the Chara braunii genome encompasses numerous genes that were previously regarded as exclusive to land plants, suggesting their acquisition prior to the colonization of terrestrial habitats. This review summarizes the established methods used to study Chara, while incorporating recent molecular data, to showcase its renewed importance as a model organism in advancing plant evolutionary developmental biology.

Interactions between stoneworts (Charales) and waterbirds.

Stoneworts (Charales) are green algae that represent an important food resource for many waterbird species in Europe and elsewhere. Browsing avian herbivores (e.g. swan, goose, duck and coot species) consume Charales plant vegetative parts, by head-dipping, up-ending or diving. A lower fibre content and longer growing season may make Charales as attractive to such herbivores as sympatric submerged higher plant species in some circumstances. Charales respond to environmental stress (e.g. drought) by producing abundant diaspores, in the form of oospores (sexual) and bulbils (asexual), both rich in starch, generating abundant food for waterbirds at critical stages in their annual migratory cycles. Waterbirds feed on these by diving (e.g. common pochard Aythya ferina and red-crested pochard Netta rufina) or by filtering from the water column (e.g. dabbling duck species), ensuring dispersal of sexually produced and vegetative diaspores locally (because of predator swamping) and remotely (through endo- and ectozoochorous dispersal by long-distance migratory waterbirds). Greater invertebrate density and diversity associated with Charales canopies enhances their attractiveness over other submerged macrophyte beds to diving predators [e.g. tufted duck Aythya fuligula, common pochard and Eurasian coot Fulica atra (hereafter coot)]. Fish fry preying on these invertebrates use such vegetation as predator cover, in turn providing prey for avian piscivores such as grebes and cormorants. Abundant Charales contribute to maintaining a transparent water column due to canopy density, nutrient effects, dampening of sedimentation/remobilisation of suspended matter and nutrients and allelopathic effects on other plants (especially phytoplankton). Shallow, relatively eutrophic waters can flip between clear-water high-biodiversity (where Charales thrive) and turbid species-poor depauperate stable states (lacking Charales). Shifts between turbid conditions and rich submerged Charales beds have profound elevating effects on aquatic diversity, to which waterbirds show rapid aggregative responses, making them ideal indicator species of ecological change; in the case of Charales specialists (such as red-crested and common pochard), indicators of the presence and abundance of these plants. Large-bodied colonial nesting birds (e.g. cormorants, gulls, heron and egrets) aggregating along lake shores contribute high N and P loadings to water bodies sensitive to such external and internal inputs and can cause local eutrophication and potential loss of Charales. Despite variation from complete seasonal removal of Charales biomass to undetectable grazing effects by herbivorous birds, evidence suggests little effect of avian grazing on biomass accumulation or the stability of community composition (under otherwise stable conditions), but we urge more research on this under-researched topic. We also lack investigations of the relative foraging profitability of different Charales organs to waterbirds and the degree of viability of gyrogonites (fertilised and calcified oospores), vegetative bulbils and plant fragments after passage through the guts of waterbirds. We especially need to understand better how much the carbonate armour of these organs affects their viability/dispersal via waterbirds and urge more research on these neglected plants and their relationships and interactions with other organisms in the aquatic biota.

Macrophytes in Inland Waters: From Knowledge to Management.

The huge biodiversity of inland waters and the many different aquatic habitats or ecosystems occurring there are particularly threatened by human impacts. In this Special Issue, ten articles have been collected that show new data on the distribution and ecology of some rare aquatic macrophytes, including both vascular plants and charophytes, but also on the use of these organisms for the monitoring, management, and restoration of wetlands.

Plastid DNA sequences and oospore characters of some European taxa of Tolypella section Tolypella (Characeae) identify five clusters, including one new cryptic Tolypella taxon from Sardinia, but they do not coincide with current morphological descriptions.

In Europe, the genus Tolypella (Characeae) comprises four to eight Tolypella taxa in sections Rothia and Tolypella that have been distinguished by vegetative morphology and gametangial characters such as antheridial size and oospore wall ornamentation. However, morphological differentiation is difficult in some cases due to overlapping and variable vegetative features, which in many cases are difficult to observe clearly. To clarify the taxonomic status of the five European taxa of Tolypella in section Tolypella, sequence data of the plastid genes atpB, rbcL and psbC for Tolypella glomerata (Desv.) Leonh., Tolypella hispanica Allen, Tolypella nidifica (O.F. Müll.) A. Braun, Tolypella normaniana (Nordst.) Nordst. and Tolypella salina Cor. were combined with data on oospore morphology, including oospore wall ornamentation. Gene sequence data identified five distinct clusters, but they were not consistent with the morphologically identified five taxa. T. glomerata consisted of some of the samples morphologically identified as T. glomerata and seven samples of T. normaniana, while the remaining T. glomerata samples clustered with specimens of unclear affiliation (Tolypella sp.). We identified two clusters of T. hispanica within the European material: cluster T. hispanica I consisted of samples from various locations, whereas the second cluster (T. hispanica II) consisted of samples of T. hispanica from Sardinia Island. The remaining cluster consisted of all the specimens that had been determined as T. salina or T. nidifica in addition to two specimens of T. normaniana. Oospore morphology was most clearly distinguishable for T. glomerata. Oospore characteristics for all other taxa were not as informative but showed some geographical and/or environmentally influenced differences, especially for T. nidifica and T. salina. Our results suggest the need to further check the different taxonomy of Tolypella sect. Tolypella in which specimens normally identified as T. glomerata might be two different taxa, T. glomerata and an unidentified taxon; T. nidifica and T. salina are not separate taxa; T. normaniana is a diminutive variant of two different Tolypella taxa; and T. hispanica comprises two different taxa, one from the Mediterranean island Sardinia.

Submergence of the filamentous Zygnematophyceae Mougeotia induces differential gene expression patterns associated with core metabolism and photosynthesis.

The streptophyte algal class Zygnematophyceae is the closest algal sister lineage to land plants. In nature, Zygnematophyceae can grow in both terrestrial and freshwater habitats and how they do this is an important unanswered question. Here, we studied what happens to the zygnematophyceaen alga Mougeotia sp., which usually occurs in permanent and temporary freshwater bodies, when it is shifted to liquid growth conditions after growth on a solid substrate. Using global differential gene expression profiling, we identified changes in the core metabolism of the organism interlinked with photosynthesis; the latter went hand in hand with measurable impact on the photophysiology as assessed via pulse amplitude modulation (PAM) fluorometry. Our data reveal a pronounced change in the overall physiology of the alga after submergence and pinpoint candidate genes that play a role. These results provide insight into the importance of photophysiological readjustment when filamentous Zygnematophyceae transition between terrestrial and aquatic habitats.