RESUMEN
PREMISE OF THE STUDY: Stinging hairs are striking examples of plant microengineering-the plant equivalent of the hypodermic syringe. The requisite mechanical properties are mostly achieved by cell wall mineralization. Stinging hairs of Urtica dioica (Urticaceae) are known to be mineralized with silica and calcium carbonate and those of Loasaceae also with calcium phosphate, but no comparative study has been provided across different taxa with stinging hairs. METHODS: Light microscopy and scanning electron microscopy (SEM) with cryo-SEM and energy-dispersive x-ray spectroscopy were used to analyze morphology and biomineralization of stinging hairs of 43 species from the families Caricaceae, Euphorbiaceae, Loasaceae, Namaceae, and Urticaceae. KEY RESULTS: Stinging hair morphology is similar across the taxa studied, in striking contrast to the divergent patterns of biomineralization. Trichome bases are mostly calcified, sometimes silicified, the shafts are mostly calcified, and the apices silicified (Urticaceae), and contain calcium phosphate (Caricaceae, Namaceae), both silica and calcium phosphate (some Loasaceae), or no minerals (Cnidoscolus, Euphorbiaceae). Some stinging hairs are superficially thinly coated with silica over a cell wall otherwise mineralized with calcium carbonate or calcium phosphate. CONCLUSIONS: Mineralization patterns are surprisingly diverse and involve three different biominerals deposited in different parts of individual trichomes with calcium phosphate a common component. The physical properties of different wall regions of the stinging trichomes are thus fine-tuned to optimize their function via modulation of wall thickness and differential element deposition. Similar function is apparently achieved through divergent wall compositions.
Asunto(s)
Minerales/metabolismo , Streptophyta/fisiología , Biomineralización , Fosfatos de Calcio/metabolismo , Pared Celular/fisiología , Pared Celular/ultraestructura , Microscopía Electrónica de Rastreo , Streptophyta/ultraestructura , Tricomas/fisiología , Tricomas/ultraestructuraRESUMEN
Two new genera (Streptosarcina and Streptofilum) and three new species (Streptosarcina arenaria, S. costaricana and Streptofilum capillatum) of streptophyte algae were detected in cultures isolated from terrestrial habitats of Europe and Central America and described using an integrative approach. Additionally, a strain isolated from soil in North America was identified as Hormidiella parvula and proposed as an epitype of this species. The molecular phylogeny based on 18S rRNA and rbcL genes, secondary structure of ITS-2, as well as the morphology of vegetative and reproductive stages, cell ultrastructure, ecology and distribution of the investigated strains were assessed. The new genus Streptosarcina forms a sister lineage to the genus Hormidiella (Klebsormidiophyceae). Streptosarcina is characterized by packet-like (sarcinoid) and filamentous thalli with true branching and a cell organization typical for Klebsormidiophyceae. Streptofilum forms a separate lineage within Streptophyta. This genus represents an easily disintegrating filamentous alga which exhibits a cell coverage of unique structure: layers of submicroscopic scales of piliform shape covering the plasmalemma and exfoliate inside the mucilage envelope surrounding cells. The implications of the discovery of the new taxa for understanding evolutionary tendencies in the Streptophyta, a group of great evolutionary interest, are discussed.
Asunto(s)
Ecosistema , Filogenia , Streptophyta/clasificación , Streptophyta/genética , América Central , Análisis por Conglomerados , ADN de Plantas/química , ADN de Plantas/genética , ADN Ribosómico/química , ADN Ribosómico/genética , ADN Espaciador Ribosómico/química , ADN Espaciador Ribosómico/genética , Europa (Continente) , Microscopía , Microscopía Electrónica de Transmisión , América del Norte , Conformación de Ácido Nucleico , ARN Ribosómico 18S/genética , Ribulosa-Bifosfato Carboxilasa/genética , Análisis de Secuencia de ADN , Microbiología del Suelo , Streptophyta/ultraestructuraRESUMEN
A bacterial endosymbiont was unexpectedly found in the "axenic" culture strain of the streptophyte green alga Mesostigma viride (NIES-995). Phylogenetic analyses based on 16S rRNA gene sequences showed that the symbiont belongs to the order Rickettsiales, specifically to the recently designated clade "Candidatus Megaira," which is closely related to the well-known Rickettsia clade. Rickettsiales bacteria of the "Ca. Megaira" clade are found in a taxonomically diverse array of eukaryotic hosts, including chlorophycean green algae, several ciliate species, and invertebrates such as Hydra. Transmission electron microscopy, fluorescence in situ hybridi-zation, and SYBR Green I staining experiments revealed that the endosymbiont of M. viride NIES-995 is rod shaped, typically occurs in clusters, and is surrounded by a halo-like structure, presumably formed by secretory substances from the bacterium. Two additional M. viride strains (NIES-296 and NIES-475), but not SAG50-1, were found to house the rickettsial endosymbiont. Analyses of strain NIES-995 transcriptome data indicated the presence of at least 91 transcriptionally active genes of symbiont origins. These include genes for surface proteins (e.g., rOmpB) that are known to play key roles in bacterial attachment onto host eukaryotes in related Rickettsia species. The assembled M. viride transcriptome includes transcripts that code for a suite of predicted algal-derived proteins, such as Ku70, WASH, SCAR, and CDC42, which may be important in the formation of the algal-rickettsial association.
Asunto(s)
Chlorophyta/microbiología , Rickettsia/fisiología , Streptophyta/microbiología , Simbiosis , Chlorophyta/genética , Chlorophyta/ultraestructura , Hibridación Fluorescente in Situ , Funciones de Verosimilitud , Microscopía Fluorescente , Anotación de Secuencia Molecular , Filogenia , ARN Mensajero/genética , ARN Mensajero/metabolismo , ARN Ribosómico 16S/genética , Subunidades Ribosómicas Pequeñas , Streptophyta/ultraestructura , Transcriptoma/genéticaRESUMEN
Key steps in evolution are often singularities. The emergence of land plants is one such case and it is not immediately apparent why. A recent analysis found that the zygnematophycean algae represent the closest relative to embryophytes. Intriguingly, many exaptations thought essential to conquer land are common among various streptophytes, but zygnematophycean algae share with land plants the transfer of a few plastid genes to the nucleus. Considering the contribution of the chloroplast to terrestrialization highlights potentially novel exaptations that currently remain unexplored. We discuss how the streptophyte chloroplast evolved into what we refer to as the embryoplast, and argue this was as important for terrestrialization by freshwater algae as the host cell-associated exaptations that are usually focused upon.
Asunto(s)
Evolución Biológica , Plastidios/fisiología , Streptophyta/fisiología , Biodiversidad , Agua Dulce , Modelos Biológicos , Streptophyta/ultraestructuraRESUMEN
The green-algal class Klebsormidiophyceae (Streptophyta), which occurs worldwide, comprises the genera Klebsormidium, Interfilum, Entransia, and Hormidiella. Ecophysiological research has so far focused on the first two genera because they are abundant in biological soil crust communities. The present study investigated the photosynthetic performances of Hormidiella attenuata and two strains of Entransia fimbriata under light, temperature, and desiccation stress. Their ultrastructure was compared using transmission electron microscopy. The two Entransia strains showed similar physiological responses. They used light more efficiently than Hormidiella, as indicated by higher oxygen production and relative electron transport rate under low light conditions, lower light saturation and compensation points, and higher maximum oxygen production during light saturation. Their requirement for low light levels explains the restriction of Entransia to dim limnetic habitats. In contrast, Hormidiella, which prefers drier soil habitats, responded to light gradients similarly to other aero-terrestrial green algae. Compared to Entransia, Hormidiella was less affected by short-term desiccation, and rehydration allowed full recovery of the photosynthetic performance. Nevertheless, both strains of Entransia coped with low water availability better than other freshwater algae. Photosynthetic oxygen production in relation to respiratory consumption was higher in low temperatures (Entransia: 5 °C, Hormidiella: 10 °C) and the ratio decreased with increasing temperatures. Hormidiella exhibited conspicuous triangular spaces in the cell wall corners, which were filled either with undulating cell wall material or with various inclusions. These structures are commonly seen in various members of Klebsormidiophyceae. The data revealed significant differences between Hormidiella and Entransia, but appropriate adaptations to their respective habitats.
Asunto(s)
Adaptación Fisiológica/fisiología , Frío , Desecación , Calor , Fotosíntesis/fisiología , Streptophyta/fisiología , Streptophyta/ultraestructura , Estrés Fisiológico , Transporte de Electrón/fisiología , Luz , Microscopía Electrónica de Transmisión , Oxígeno/metabolismo , Streptophyta/clasificaciónRESUMEN
Two newly isolated strains of green algae from alpine regions were compared physiologically at different culture ages (1, 6, 9 and 15 months). The strains of Zygnema sp. were from different altitudes ('Saalach' (S), 440 m above sea level (a.s.l.), SAG 2419 and 'Elmau-Alm' (E-A), 1,500 m a.s.l., SAG 2418). Phylogenetic analysis of rbcL sequences grouped the strains into different major subclades of the genus. The mean diameters of the cells were 23.2 µm (Zygnema S) and 18.7 µm (Zygnema E-A) but were reduced significantly with culture age. The photophysiological response between the strains differed significantly; Zygnema S had a maximal relative electron transport rate (rETR max) of 103.4 µmol electrons m(-2) s(-1), Zygnema E-A only 61.7 µmol electrons m(-2) s(-1), and decreased significantly with culture age. Both strains showed a low-light adaption and the absence of strong photoinhibition up to 2,000 µmol photons m(-2) s(-1). Photosynthetic oxygen production showed similar results (P max Zygnema S, 527.2 µmol O2 h(-1) mg(-1) chlorophyll (chl.) a, Zygnema E-A, 390.7 µmol O2 h(-1) mg(-1) chl. a); the temperature optimum was at 35 °C for Zygnema S and 30 °C for Zygnema E-A. Increasing culture age moreover leads to the formation of pre-akinetes, which accumulate storage products as revealed by light and transmission electron microscopy. Desiccation at 84 % relative air humidity (RH) lead to a reduction of the effective quantum yield of photosystem II (PSII) (ΔFv/Fm') to zero between 90 to 120 min (Zygnema S) and between 30 to 60 min (Zygnema E-A), depending on the culture age. A partial recovery of ΔFv/Fm' was only observed in older cultures. We conclude that pre-akinetes are crucial for the aeroterrestrial lifestyle of Zygnema.
Asunto(s)
Fotosíntesis , Streptophyta/metabolismo , Adaptación Fisiológica , Deshidratación , Transporte de Electrón , Datos de Secuencia Molecular , Oxígeno/metabolismo , Filogenia , Análisis de Secuencia de ADN , Streptophyta/genética , Streptophyta/ultraestructura , Estrés FisiológicoRESUMEN
Antarctic algae play a fundamental role in polar ecosystem thanks to their ability to grow in an extreme environment characterized by low temperatures and variable illumination. Here, for prolonged periods, irradiation is extremely low and algae must be able to harvest light as efficiently as possible. On the other side, at low temperatures even dim irradiances can saturate photosynthesis and drive to the formation of reactive oxygen species. Colonization of this extreme environment necessarily required the optimization of photosynthesis regulation mechanisms by algal organisms. In order to investigate these adaptations we analyzed the time course of physiological and morphological responses to different irradiances in Koliella antarctica, a green microalga isolated from Ross Sea (Antarctica). Koliella antarctica not only modulates cell morphology and composition of its photosynthetic apparatus on a long-term acclimation, but also shows the ability of a very fast response to light fluctuations. Koliella antarctica controls the activity of two xanthophyll cycles. The first, involving lutein epoxide and lutein, may be important for the growth under very low irradiances. The second, involving conversion of violaxanthin to antheraxanthin and zeaxanthin, is relevant to induce a fast and particularly strong non-photochemical quenching, when the alga is exposed to higher light intensities. Globally K. antarctica thus shows the ability to activate a palette of responses of the photosynthetic apparatus optimized for survival in its natural extreme environment.
Asunto(s)
Aclimatación/efectos de la radiación , Fotosíntesis/efectos de la radiación , Streptophyta/efectos de la radiación , Adaptación Fisiológica , Frío , Ambiente , Luz , Luteína/metabolismo , Streptophyta/fisiología , Streptophyta/ultraestructura , Xantófilas/metabolismo , Zeaxantinas/metabolismoRESUMEN
BACKGROUND: Filamentous Zygnematophyceae are typical components of algal mats in the polar hydro-terrestrial environment. Under field conditions, they form senescent vegetative cells, designated as pre-akinetes, which are tolerant to desiccation and osmotic stress. KEY FINDINGS: Pre-akinete formation and desiccation tolerance was investigated experimentally under monitored laboratory conditions in four strains of Arctic and Antarctic isolates with vegetative Zygnema sp. morphology. Phylogenetic analyses of rbcL sequences revealed one Arctic strain as genus Zygnemopsis, phylogenetically distant from the closely related Zygnema strains. Algae were cultivated in liquid or on solidified medium (9 weeks), supplemented with or lacking nitrogen. Nitrogen-free cultures (liquid as well as solidified) consisted of well-developed pre-akinetes after this period. Desiccation experiments were performed at three different drying rates (rapid: 10% relative humidity, slow: 86% rh and very slow); viability, effective quantum yield of PS II, visual and ultrastructural changes were monitored. Recovery and viability of pre-akinetes were clearly dependent on the drying rate: slower desiccation led to higher levels of survival. Pre-akinetes survived rapid drying after acclimation by very slow desiccation. CONCLUSIONS: The formation of pre-akinetes in polar Zygnema spp. and Zygnemopsis sp. is induced by nitrogen limitation. Pre-akinetes, modified vegetative cells, rather than specialized stages of the life cycle, can be hardened by mild desiccation stress to survive rapid drying. Naturally hardened pre-akinetes play a key role in stress tolerance and dispersal under the extreme conditions of polar regions, where sexual reproduction and production of dormant stages is largely suppressed.
Asunto(s)
Nitrógeno/metabolismo , Streptophyta/metabolismo , Regiones Antárticas , Regiones Árticas , Secuencia de Bases , ADN de Plantas/análisis , Desecación , Ecosistema , Microscopía Electrónica de Transmisión , Datos de Secuencia Molecular , Presión Osmótica , Complejo de Proteína del Fotosistema II/química , Complejo de Proteína del Fotosistema II/metabolismo , Filogenia , Reacción en Cadena de la Polimerasa , Streptophyta/clasificación , Streptophyta/ultraestructuraRESUMEN
BACKGROUND: Water loss has significant effects on physiological performance and survival rates of algae. However, despite the prominent presence of aeroterrestrial algae in terrestrial habitats, hardly anything is known about the molecular events that allow aeroterrestrial algae to survive harsh environmental conditions. We analyzed the transcriptome and physiology of a strain of the alpine aeroterrestrial alga Klebsormidium crenulatum under control and strong desiccation-stress conditions. PRINCIPAL FINDINGS: For comparison we first established a reference transcriptome. The high-coverage reference transcriptome includes about 24,183 sequences (1.5 million reads, 636 million bases). The reference transcriptome encodes for all major pathways (energy, carbohydrates, lipids, amino acids, sugars), nearly all deduced pathways are complete or missing only a few transcripts. Upon strong desiccation, more than 7000 transcripts showed changes in their expression levels. Most of the highest up-regulated transcripts do not show similarity to known viridiplant proteins, suggesting the existence of some genus- or species-specific responses to desiccation. In addition, we observed the up-regulation of many transcripts involved in desiccation tolerance in plants (e.g. proteins similar to those that are abundant in late embryogenesis (LEA), or proteins involved in early response to desiccation ERD), and enzymes involved in the biosynthesis of the raffinose family of oligosaccharides (RFO) known to act as osmolytes). Major physiological shifts are the up-regulation of transcripts for photosynthesis, energy production, and reactive oxygen species (ROS) metabolism, which is supported by elevated cellular glutathione content as revealed by immunoelectron microscopy as well as an increase in total antiradical power. However, the effective quantum yield of Photosystem II and CO2 fixation decreased sharply under the applied desiccation stress. In contrast, transcripts for cell integrative functions such as cell division, DNA replication, cofactor biosynthesis, and amino acid biosynthesis were down-regulated. SIGNIFICANCE: This is the first study investigating the desiccation transcriptome of a streptophyte green alga. Our results indicate that the cellular response is similar to embryophytes, suggesting that embryophytes inherited a basic cellular desiccation tolerance from their streptophyte predecessors.
Asunto(s)
Adaptación Fisiológica/genética , Desecación , Streptophyta/genética , Streptophyta/fisiología , Transcriptoma/genética , Ácido Ascórbico/metabolismo , Carotenoides/biosíntesis , Sequías , Regulación de la Expresión Génica , Glutatión/metabolismo , Complejo de Proteína del Fotosistema II/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Streptophyta/inmunología , Streptophyta/ultraestructuraRESUMEN
Plant species collected from various climatic zones and stressed in vitro at various temperatures reveal changes in cellular ultrastructure which are in accordance with the climate at their sampling sites. This observation initiated the investigation to establish if stress at different temperatures may cause diverse extents of changes in the ultrastructure of microalgal strains originating from different geographic zones. The study revealed that the six Cosmarium strains demonstrated ultrastructural characteristics that were consistent with their source location under optimal, low and high temperature conditions, pointing to their preference to specific climatic niches. Interestingly, chloroplasts of all of the Cosmarium strains correspond to a sun-adapted type, which is concomitant with earlier statements that these strains are rendered as high-light adapted algae. The Cosmarium strains developed multiple ultrastructural responses which enabled them to cope with excessive temperatures, occasionally occurring in desmid natural habitats. The appearance of cubic membranes and increased number of plastoglobules may represent the first line in protection against high-temperature stress, which is accompanied by the alteration of protein synthesis and the appearance of stress granules in order to preserve cell homeostasis. However, the prolonged warm- or cold-temperature stress obviously initiated the programmed cell death, as concluded from the appearance of several ultrastructural features observed in all of the Cosmarium strains. The fair acclimation possibilities and the ability to undergo programmed cell death in order to save the population, certainly favor the cosmopolitan distribution of the genus Cosmarium.
Asunto(s)
Geografía , Streptophyta/fisiología , Streptophyta/ultraestructura , Estrés Fisiológico , Temperatura , Cloroplastos/efectos de la radiación , Cloroplastos/ultraestructura , Respuesta al Choque Térmico/efectos de la radiación , Indoles/metabolismo , Luz , Especificidad de la Especie , Streptophyta/crecimiento & desarrollo , Streptophyta/efectos de la radiación , Estrés Fisiológico/efectos de la radiaciónRESUMEN
Ultraviolet (UV) radiation has become an important stress factor in polar regions due to anthropogenically induced ozone depletion. Although extensive research has been conducted on adaptations of polar organisms to this stress factor, few studies have focused on semi-terrestrial algae so far, in spite of their apparent vulnerability. This study investigates the effect of UV on two semi-terrestrial arctic strains (B, G) and one Antarctic strain (E) of the green alga Zygnema, isolated from Arctic and Antarctic habitats. Isolates of Zygnema were exposed to experimentally enhanced UV A and B (predominant UV A) to photosynthetic active radiation (PAR) ratio. The pigment content, photosynthetic performance and ultrastructure were studied by means of high-performance liquid chromatography (HPLC), chlorophyll a fluorescence and transmission electron microscopy (TEM). In addition, phylogenetic relationships of the investigated strains were characterised using rbcL sequences, which determined that the Antarctic isolate (E) and one of the Arctic isolates (B) were closely related, while G is a distinct lineage. The production of protective phenolic compounds was confirmed in all of the tested strains by HPLC analysis for both controls and UV-exposed samples. Moreover, in strain E, the content of phenolics increased significantly (p = 0.001) after UV treatment. Simultaneously, the maximum quantum yield of photosystem II photochemistry significantly decreased in UV-exposed strains E and G (p < 0.001), showing a clear stress response. The phenolics were most probably stored at the cell periphery in vacuoles and cytoplasmic bodies that appear as electron-dense particles when observed by TEM after high-pressure freeze fixation. While two strains reacted moderately on UV exposure in their ultrastructure, in strain G, damage was found in chloroplasts and mitochondria. Plastidal pigments and xanthophyll cycle pigments were investigated by HPLC analysis; UV A- and UV B-exposed samples had a higher deepoxidation state as controls, particularly evident in strain B. The results indicate that phenolics are involved in UV protection of Zygnema and also revealed different responses to UV stress across the three strains, suggesting that other protection mechanisms may be involved in these organisms.
Asunto(s)
Fenoles/análisis , Streptophyta/efectos de la radiación , Streptophyta/ultraestructura , Rayos Ultravioleta , Regiones Antárticas , Regiones Árticas , Clorofila/análisis , Clorofila A , Cromatografía Líquida de Alta Presión , ADN de Plantas/genética , Microscopía Electrónica de Transmisión , Fotosíntesis/efectos de la radiación , Complejo de Proteína del Fotosistema II/efectos de la radiación , Filogenia , Streptophyta/química , Xantófilas/análisisRESUMEN
Members of the cosmopolitan green algal genus Klebsormidium (Klebsormidiales, Streptophyta) are typical components of terrestrial microbiotic communities such as biological soil crusts, which have many important ecological functions. In the present study, Klebsormidium dissectum (Gay) Ettl & Gärtner was isolated from a high alpine soil crust in the Tyrolean Alps, Austria. Physiological performance in terms of growth and photosynthesis was investigated under different controlled abiotic conditions and compared with ultrastructural changes under the treatments applied. K. dissectum showed very low light requirements as reflected in growth patterns and photosynthetic efficiency. Increasing temperatures from 5°C to 40°C led to different effects on respiratory oxygen consumption and photosynthetic oxygen evolution. While at low temperatures (5-10°C), respiration was not detectable or on a very low level, photosynthesis was relatively high, Reversely, at the highest temperature, respiration was unaffected, and photosynthesis strongly inhibited pointing to strong differences in temperature sensitivity between both physiological processes. Although photosynthetic performance of K. dissectum was strongly affected under short-term desiccation and recovered only partly after rehydration, this species was capable to survive even 3 weeks at 5% relative air humidity. K. dissectum cells have a cell width of 5.6 ± 0.3 µm and a cell length of 8.4 ± 2.0 µm. Desiccated cells showed a strongly reduced cell width (46% of control) and cell length (65% of control). In addition, in desiccated cells, fewer mitochondria were stained by DIOC(6), and damaged plasma membranes were detected by FM 1-43 staining. High-pressure freeze fixation as well as chemical fixation allowed visualizing ultrastructural changes caused by desiccation. In such cells, the nucleus and chloroplast were still visibly intact, but the extremely thin cell walls (75-180 nm) were substantially deformed. The cytoplasm appeared electron dense and mitochondria were altered. Although K. dissectum can be characterized as euryoecious species, all ecophysiological and ultrastructural data indicate susceptibility to desiccation. However, the steadily occurring fragmentation of filaments into smaller units leads to improved self protection and thus may represent a life strategy to better survive longer periods of drought in exposed alpine soil crusts.
Asunto(s)
Adaptación Fisiológica , Fotosíntesis , Streptophyta/metabolismo , Streptophyta/ultraestructura , Deshidratación , Sequías , Luz , Suelo , TemperaturaRESUMEN
The osmotic potential and effects of plasmolysis were investigated in two different Klebsormidium strains from alpine habitats by incubation in 300-2,000 (3,000) mM sorbitol. Several members of this genus were previously found to tolerate desiccation in the vegetative state yet information was lacking on the osmotic potentials of these algae. The strains were morphologically determined as Klebsormidium crenulatum and Klebsormidium nitens. These species belong to distinct clades, as verified by phylogenetic analysis of the rbcL gene. K. crenulatum is part of to the K. crenulatum/mucosum ('F' clade) and K. nitens of the 'E2' clade. Plasmolysis occurred in K. crenulatum at 800 mM sorbitol (961 mOsmol kg(-1), Ψ = -2.09 MPa) and in K. nitens at 600 mM sorbitol (720 mOsmol kg(-1), Ψ = -1.67 MPa). These are extraordinarily high osmotic values (very negative osmotic potentials) compared with values reported for other green algae. In K. crenulatum, the maximum photosynthetic rate (Pmax) in the light-saturated range was 116 µmol O(2) h(-1) mg(-1) chl a. Incubation in 1,000 mM sorbitol decreased Pmax to 74.1% of the initial value, whereas 2,000 mM sorbitol (Ψ = -5.87 MPa) lead to an almost complete loss of oxygen production. In K. nitens, Pmax was 91 µmol O(2) h(-1) mg(-1) chl a under control conditions and incubation in 800 mM sorbitol did not decrease Pmax, 2,000 mM sorbitol decreased Pmax only to about 62.6% of the initial value whereas 3,000 mM sorbitol stopped oxygen evolution. This indicated a broader amplitude for photosynthesis in the examined strain of K. nitens. Control samples and samples plasmolysed for 3 h in 800 mM sorbitol (K. nitens), 1,000 mM sorbitol (K. crenulatum), or 2,000 mM sorbitol were investigated by transmission electron microscopy after chemical or high-pressure freeze fixation. In cells undergoing plasmolysis the protoplasts were retracted from the cell wall, the cytoplasm appeared dense, vacuoles were small and fragmented, and the cytoplasm was filled with ribosomes. Thin cytoplasmic strands were connected to the cell wall; 2,000 mM sorbitol increased the effect. The content of soluble carbohydrates in these two strains was investigated by HPLC, as this is one known mechanism for cells to maintain high osmotic pressure of the cytosol. Both Klebsormidium species contained diverse soluble carbohydrates, including a dominant mixed peak of unidentified oligosaccharides, and more minor amounts of raffinose, sucrose, glucose, xylose, galactose, mannose, inositol, fructose, glycerol, mannitol, and sorbitol. The total content of soluble carbohydrates was approximately 1.2% of the dry weight, indicating that this is not a major factor contributing to the high osmotic potential in these strains of Klebsormidium.
Asunto(s)
Streptophyta/ultraestructura , Deshidratación , Higroscópicos/química , Monosacáridos/aislamiento & purificación , Monosacáridos/metabolismo , Oligosacáridos/aislamiento & purificación , Oligosacáridos/metabolismo , Presión Osmótica , Oxígeno/metabolismo , Fotosíntesis , Filogenia , Protoplastos/fisiología , Ribulosa-Bifosfato Carboxilasa/genética , Análisis de Secuencia de ADN , Sorbitol/química , Streptophyta/genética , Streptophyta/metabolismo , Estrés FisiológicoRESUMEN
Simple sequence repeats (SSRs) or microsatellites constitute a countable portion of genomes. However, the significance of SSRs in organelle genomes has not been completely understood. The availability of organelle genome sequences allows us to understand the organization of SSRs in their genic and intergenic regions. In the current study we surveyed the patterns of SSRs in mitochondrial genomes of different taxa of plants. A total of 16 mitochondrial genomes, from algae to angiosperms, have been considered to analyze the pattern of simple sequence repeats present in them. Based on study, the mononucleotide repeats of A/T were found to be more prevalent in mitochondrial genomes over other repeat types. The dinucleotides repeats, TA/AT, were the second most numerous, whereas tri-, tetra-, and pentanucleotide repeats were in less number and present in intronic or intergenic portions only. Mononucleotide repeats prevailed in protein-coding exonic portions of all organisms. These results indicates that microsatellite pattern in mitochondrial genomes is different from nuclear genomes and also focuses on organization and diversity at SSR locuses in mitochondrial genomes. This is the novel report of microsatellite polymorphism in plant mitochondrion on whole genome level.
Asunto(s)
Genoma Mitocondrial , Genoma de Planta , Repeticiones de Microsatélite , Mitocondrias/genética , Streptophyta/genética , ADN Mitocondrial/química , Análisis de Secuencia de ADN , Streptophyta/ultraestructuraRESUMEN
A detailed comparative examination of microtubule (MT) organization in interphase and dividing cells of Uronema sp., Klebsormidium flaccidum, K. subtilissimum, Stichococcus bacillaris and S. chloranthus was made using tubulin immunofluorescence and transmission electron microscopy (TEM). During interphase all the species bear a well-organized cortical MT system, consisting of parallel bundles with different orientations. In Uronema sp. the cortical MT bundles are longitudinally oriented, whereas in the other species they are in transverse orientation to the axis of the cells. Considerable differences in MT organization were also observed during stages of mitosis, mainly preprophase, as well as cytokinesis. In Uronema sp., a particular radial MT assembly is organized during preprophase-early prophase, which was not observed in the other species. In Stichococcus a fine MT ring surrounded the nucleus during preprophase and prophase. An MT ring, together with single cytoplasmic MTs, was also found associated with the developing diaphragm during cytokinesis in Stichococcus. A phycoplast participates in cytokinesis in Uronema sp., but not in the other species. In Uronema sp. the centrosome functions as a microtubule organizing center (MTOC) during mitosis, but not during interphase and cytokinesis. The phylogenetic significance of these differences is discussed in combination with SSU/ITS sequencing and other, existing molecular data.
Asunto(s)
Chlorophyta/ultraestructura , Cilióforos/ultraestructura , Centro Organizador de los Microtúbulos/ultraestructura , Streptophyta/ultraestructura , Chlorophyta/clasificación , Chlorophyta/citología , Chlorophyta/genética , Cilióforos/clasificación , Cilióforos/citología , Cilióforos/genética , Técnica del Anticuerpo Fluorescente , Microscopía Electrónica de Transmisión , Datos de Secuencia Molecular , Filogenia , Streptophyta/clasificación , Streptophyta/citología , Streptophyta/genéticaRESUMEN
The core of plant physiology is a set of functional solutions to a tradeoff between CO(2) acquisition and water loss. To provide an important evolutionary perspective on how the earliest land plants met this tradeoff, we constructed a mathematical model (constrained geometrically with measurements of fossils) of the hydraulic resistance of Asteroxylon, an Early Devonian plant. The model results illuminate the water transport physiology of one of the earliest vascular plants. Results show that Asteroxylon's vascular system contains cells with low hydraulic resistances; these resistances are low because cells were covered by scalariform pits, elliptical structures that permit individual cells to have large areas for water to pass from one cell to another. Asteroxylon could move a large amount of water quickly given its large pit areas; however, this would have left these plants particularly vulnerable to damage from excessive evapotranspiration. These results highlight a repeated pattern in plant evolution, wherein the evolution of highly conductive vascular tissue precedes the appearance of adaptations to increase water transport safety. Quantitative insight into the vascular transport of Asteroxylon also allows us to reflect on the quality of CO(2) proxy estimates based on early land plant fossils. Because Asteroxylon's vascular tissue lacked any safety features to prevent permanent damage, it probably used stomatal abundance and behavior to prevent desiccation. If correct, low stomatal frequencies in Asteroxylon reflect the need to limit evapotranspiration, rather than adaptation to high CO(2) concentrations in the atmosphere. More broadly, methods to reveal and understand water transport in extinct plants have a clear use in testing and bolstering fossil plant-based paleoclimate proxies.