Food preference of freshwater invertebrates: comparing fresh and decomposed angiosperm and a filamentous alga

1. Fresh and decomposed Mougeotia sp. (a filamentous green alga) and Elodea nuttallii (a vascular plant) were offered as food to three species of aquatic macroinvertebrates (Lymnaea peregra, Asellus meridianus and Endochironomus albipennis) to test: (i) if filamentous algae are preferred to aquatic higher plants (hereafter, called ‘macrophytes’) and (ii), as is known for higher plants, if decomposition also results in greater palatability of filamentous algae. 2. Compared with the alga, the macrophyte in both states was of higher nutritional value. Conditioning improved the nutritional value of both food types, but especially of the macrophyte. 3. Both fresh alga and fresh macrophyte were eaten little by all animals except A. meridianus feeding upon the macrophyte. Consumption was higher for both plants in their decomposed state. However, L. peregra consumed more decomposed macrophyte than the decomposed alga. Both decomposed plants were eaten most by E. albipennis followed by A. meridianus and L. peregra. 4. Digestibility of both plants, but especially of the macrophyte, increased significantly after decomposition. The assimilation efficiencies of all animals on the fresh E. nuttallii were higher than on fresh Mougeotia sp. After decomposition, the efficiency increased significantly only on the alga. Consequently, both decomposed plants were assimilated with similar efficiency by all test animals. 5. Amongst aquatic macrophytes, the increase of their consumption and digestibility upon decomposition has hitherto been known only for vascular plants but not for filamentous algae.     

Sucrose phosphate phosphatase in the green alga <Emphasis Type="Italic">Klebsormidium flaccidum</Emphasis> (Streptophyta) lacks an extensive C-terminal domain and differs from that of land plants

Previously, it was reported that like land plants, the green alga Klebsormidium flaccidum (Streptophyta) accumulates sucrose during cold acclimation (Nagao et al. Plant Cell Environ 31:872–885, 2008), suggesting that synthesis of sucrose could enhance the freezing tolerance of this alga. Because sucrose phosphate phosphatase (SPP; EC 3.1.3.24) is a key enzyme in the sucrose synthesis pathway in plants, we analyzed the SPP gene in K. flaccidum (KfSPP, GenBank accession number AB669024) to clarify its role in sucrose accumulation. As determined from its deduced amino acid sequence, KfSPP contains the N-terminal domain that is characteristic of the L-2-haloacid-dehalogenase family of phosphatases/hydrolases (the HAD phosphatase domain). However, it lacks the extensive C-terminal domain found in SPPs of land plants. Database searches revealed that the SPPs in cyanobacteria also lack the C-terminal domain. In addition, the green alga Coccomyxa (Chlorophyta) and K. flaccidum, which are closely related to land plants, have cyanobacterial-type SPPs, while Chlorella (Chlorophyta) has a land plant-type SPP. These results demonstrate that even K. flaccidum (Streptophyta), as a recent ancestor of land plants, has the cyanobacterial-type SPP lacking the C-terminal domain. Because SPP and sucrose phosphate synthase (SPS) catalyze sequential reactions in sucrose synthesis in green plant cells and the lack of the C-terminal domain in KfSPP is predicted to decrease its activity, the interaction between decreased KfSPP activity and SPS activity may alter sucrose synthesis during cold acclimation in K. flaccidum.     

The Complete Mitochondrial Genome Sequence of the Hornwort <Emphasis Type="Italic">Megaceros</Emphasis><Emphasis Type="Italic">aenigmaticus</Emphasis> Shows a Mixed Mode of Conservative Yet Dynamic Evolution in Early Land Plant Mitochondrial Genomes

Land plants possess some of the most unusual mitochondrial genomes among eukaryotes. However, in early land plants these genomes resemble those of green and red algae or early eukaryotes. The question of when during land plant evolution the dramatic change in mtDNAs occurred remains unanswered. Here we report the first completely sequenced mitochondrial genome of the hornwort, Megaceros aenigmaticus, a member of the sister group of vascular plants. It is a circular molecule of 184,908 base pairs, with 32 protein genes, 3 rRNA genes, 17 tRNA genes, and 30 group II introns. The genome contains many genes arranged in the same order as in those of a liverwort, a moss, several green and red algae, and Reclinomonas americana, an early-branching eukaryote with the most ancestral form of mtDNA. In particular, the gene order between mtDNAs of the hornwort and Physcomitrella patens (moss) differs by only 8 inversions and translocations. However, the hornwort mtDNA possesses 4 derived features relative to green alga mtDNAs—increased genome size, RNA editing, intron gains, and gene losses—which were all likely acquired during the origin and early evolution of land plants. Overall, this genome and those of other 2 bryophytes show that mitochondrial genomes in early land plants, unlike their seed plant counterparts, exhibit a mixed mode of conservative yet dynamic evolution. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users. Libo Li and Bin Wang contributed equally to this work.     

Are the photosynthetic membranes of cryptophyte algae inside out?

Summary Cryptomonads are unicellular algae with a unique photosynthetic apparatus, both in structure and pigment composition. A cryptomonad,Rhodomonas lens (R. lens), was studied by conventional electron microscopy, freeze-fracture, and freeze-etch in order to determine whether the thylakoids of this alga are everted with respect to those of other plants, as has been postulated (Gantt et al. 1971,Gantt 1979, 1980) as a means to compensate for the location of the cryptomonad light-harvesting apparatus on the opposite side of the thylakoid membrane from that of related algae. We have characterized the thylakoids of this alga and conclude that they are not everted, but are oriented in the same manner as those of other algae and green plants. Implications for energy transfer are discussed.     

Evolution of Plant-Like Crystalline Storage Polysaccharide in the Protozoan Parasite Toxoplasma gondii Argues for a Red Alga Ancestry

Single-celled apicomplexan parasites are known to cause major diseases in humans and animals including malaria, toxoplasmosis, and coccidiosis. The presence of apicoplasts with the remnant of a plastid-like DNA argues that these parasites evolved from photosynthetic ancestors possibly related to the dinoflagellates. Toxoplasma gondii displays amylopectin-like polymers within the cytoplasm of the dormant brain cysts. Here we report a detailed structural and comparative analysis of the Toxoplasma gondii, green alga Chlamydomonas reinhardtii, and dinoflagellate Crypthecodinium cohnii storage polysaccharides. We show Toxoplasma gondii amylopectin to be similar to the semicrystalline floridean starch accumulated by red algae. Unlike green plants or algae, the nuclear DNA sequences as well as biochemical and phylogenetic analysis argue that the Toxoplasma gondii amylopectin pathway has evolved from a totally different UDP-glucose-based metabolism similar to that of the floridean starch accumulating red alga Cyanidioschyzon merolae and, to a lesser extent, to those of glycogen storing animals or fungi. In both red algae and apicomplexan parasites, isoamylase and glucan–water dikinase sequences are proposed to explain the appearance of semicrystalline starch-like polymers. Our results have built a case for the separate evolution of semicrystalline storage polysaccharides upon acquisition of photosynthesis in eukaryotes.This article contains online-only supplementary material.Reviewing Editor:Dr. Patrick Keeling     

Chloroplast transit peptides from the green alga Chlamydomonas reinhardtii share features with both mitochondrial and higher plant chloroplast presequences

Chloroplast transit peptides from the green alga Chlamydomonas reinhardtii have been analyzed and compared with chloroplast transit peptides from higher plants and mitochondrial targeting peptides from yeast, Neurospora and higher eukaryotes. In terms of length and amino acid composition, chloroplast transit peptides from C. reinhardtii are more similar to mitochondrial targetting peptides than to chloroplast transit peptides from higher plants. They also contain the potential amphiphilic α-helix characteristic of mitochondrial presequences. However, in similarity with chloroplast transit peptides from higher plants, they contain a C-terminal region with the potential to form an amphiphilic β-strand. As in higher plants, transit peptides that route proteins to the thylakoid lumen consist of an N-tenninal domain similar to stroma-targeting transit peptides attached to a C-terminal apolar domain that share many characteristics with secretory signal peptides.     

PHOTOSYNTHETIC PIGMENT COMPOSITION IN THE PRIMITIVE GREEN ALGA MESOSTIGMA VIRIDE (PRASINOPHYCEAE): PHYLOGENETIC AND EVOLUTIONARY IMPLICATIONS1

The photosynthetic pigment composition of Mesostigma viride Lauterborn, a primitive green alga, was determined. This alga contained chl a and b, lycopene, lutein, siphonaxanthin, γ‐carotene, β‐carotene, antheraxanthin, violaxanthin, neoxanthin, and two novel carotenoid fatty acid esters, siphonaxanthin C12:0 ester and siphonaxanthin C14:0 ester. The esters were saturated, whereas all previously identified siphonaxanthin and loroxanthin esters have been mono‐unsaturated (trans‐Δ2). Neoxanthin was the all‐trans form. This is the first such case detected in the chloroplasts of green plants. The 9′‐cis form of neoxanthin is believed to be universally present in the chloroplasts of green plants (Streptophyta and Chlorophyta) and is a precursor of abscisic acid. However, the 9′‐cis form was not found in M. viride. Based on these results, we discuss the phylogenetic implications and early evolution of the antenna pigment system in green plants.     

Mechanism of plasmodesmata formation in characean algae in relation to evolution of intercellular communication in higher plants

It is generally accepted that higher plants evolved from ancestral forms of the modern charophytes. For this reason, we chose the characean alga, Chara corallina Klein ex Willd., em. R.D.W. (C. australis R. Br.), to determine whether this transition species produces plasmodesmata in a manner analogous to higher plants. As with higher plants and unlike most green algae, Chara utilizes a phragmoplast for cell division; however, in contrast with the situation in both lower and higher vascular plants, the developing cell plate and newly formed cell wall were found to be completely free of plasmodesmata. Only when the daughter cells had separated completely were plasmodesmata formed across the division wall. Presumably, highly localized activity of wall-degrading (or loosening) enzymes inserted into the plasma membrane play a central role in this process. In general appearance characean plasmodesmata are similar to those of higher plants with the notable exception that they lack an appressed endoplasmic reticulum. Further secondary modifications in plasmodesmal structure were found to occur as a function of cell development, giving rise to highly branched plasmodesmata in mature cell walls. These findings are discussed in terms of the evolution of the mechanism for plasmodesmata formation in algae and higher plants.This work was supported in part by National Foundation grant No. DCB-9016756 (W.J.L.). We thank the Electron Microscopy Center of Washington State University and the Zoology Department, University of California, Davis, for the use of their microscopy facilities.     

Evolutionary patterns in auxin action

This review represents the first effort ever to survey the entire literature on auxin (indole-3-acetic acid, IAA) action in all plants, with special emphasis on the green plant lineage, including charophytes (the green alga group closest to the land plants), bryophytes (the most basal land plants), pteridophytes (vascular non-seed plants), and seed plants. What emerges from this survey is the surprising perspective that the physiological mechanisms for regulating IAA levels and many IAA-mediated responses found in seed plants are also present in charophytes and bryophytes, at least in nascent forms. For example, the available evidence suggests that the apical regions of both charophytes and liverworts synthesize IAA via a tryptophan-independent pathway, with IAA levels being regulated via the balance between the rates of IAA biosynthesis and IAA degradation. The apical regions of all the other land plants utilize the same class of biosynthetic pathway, but they have the potential to utilize IAA conjugation and conjugate hydrolysis reactions to achieve more precise spatial and temporal control of IAA levels. The thallus tips of charophytes exhibit saturable IAA influx and efflux carriers, which are apparently not sensitive to polar IAA transport inhibitors. By contrast, two divisions of bryophyte gametophytes and moss sporophytes are reported to carry out polar IAA transport, but these groups exhibit differing sensitivities to those inhibitors. Although the IAA regulation of charophyte development has received almost no research attention, the bryophytes manifest a wide range of developmental responses, including tropisms, apical dominance, and rhizoid initiation, which are subject to IAA regulation that resembles the hormonal control over corresponding responses in seed plants. In pteridophytes, IAA regulates root initiation and vascular tissue differentiation in a manner also very similar to its effects on those processes in seed plants. Thus, it is concluded that the seed plants did not evolve de novo mechanisms for mediating IAA responses, but have rather modified pre-existing mechanisms already operating in the early land plants. Finally, this paper discusses the encouraging prospects for investigating the molecular evolution of auxin action.     

PRODUCTS OF PHOTOSYNTHESIS IN CAULERPA SIMPLICIUSCULA (CHLOROPHYCEAE)1

The kinetics of carbon flow through various metabolites of Caulerpa simpliciuscula C. Ag., a siphonaceous green alga, are described. The pattern of C fixation in this alga differs from those observed in higher plants and other green algae in a number of respects. After 40 min incubation, soluble β-linked polyglucans are the most important labelled carbohydrates. Sucrose, although delectable, is never a major recipient of label and does not label until after other oligosaccharide fractions. The sugar monophosphates contain a large proportion of the ¹⁴C assimilated even after 40 min and behave not only as metabolic intermediates, but also as storage pools. The pattern of ammo acid labelling resembles that reported for other species of green algae in that alanine becomes labelled most rapidly, followed by glycine, and much later by glutamate, glutamine and asparagine. These labelling patterns are discussed in the light, of what is known of patterns of C flow in other algae.     

An update on carotenoid biosynthesis in algae: phylogenetic evidence for the existence of two classes of phytoene synthase

Carotenoids play crucial roles in structure and function of the photosynthetic apparatus of bacteria, algae, and higher plants. The entry-step reaction to carotenoid biosynthesis is catalyzed by the phytoene synthase (PSY), which is structurally and functionally related in all organisms. A comparative genomic analysis regarding the PSY revealed that the green algae Ostreococcus and Micromonas possess two orthologous copies of the PSY genes, indicating an ancient gene duplication event that produced two classes of PSY in algae. However, some other green algae (Chlamydomonas reinhardtii, Chlorella vulgaris, and Volvox carteri), red algae (Cyanidioschyzon merolae), diatoms (Thalassiosira pseudonana and Phaeodactylum tricornutum), and higher plants retained only one class of the PSY gene whereas the other gene copy was lost in these species. Further, similar to the situation in higher plants recent gene duplications of PSY have occurred for example in the green alga Dunaliella salina/bardawil. As members of the PSY gene families in some higher plants are differentially regulated during development or stress, the discovery of two classes of PSY gene families in some algae suggests that carotenoid biosynthesis in these algae is differentially regulated in response to development and environmental stress as well.     

SEQUENCE AND PHYLOGENY OF THE PSAB GENE OF PYLAIELLA LITTORALIS(PHAEOPHYTA)1

The psaB gene codes for one of two highly conserved P700 chlorophyll a apoproteins of photosystem I. This gene was cloned from the brown alga, Pylaiella littoralis (L.) Kjellm., and its primary sequence was determined. The inferred amino acid sequence of the P. littoralis protein was compared to homologous sequences from land plants, green algae, and a cyanobacterium. The psaB protein sequence is very conserved in all the examined taxa, and an unrooted phylogenetic tree, generated from a distance matrix, shows that the P. littoralis gene is closer to that of the cyanobacterium Synechocossus sp. PCC 7002 than are those of green algae, land plants, and Euglena gracilis.     

Isolation of protoplasts and ion channel recording of plasma membrane patches and whole-cell recordings of the marine alga, <Emphasis Type="Italic">Ulva pertusa</Emphasis> (Chlorophyta)

Whole-cell patch-clamp techniques were used to study ion channels of a marine alga. High quality protoplasts suitable for electrophysiological studies were isolated from the green marine alga, Ulva pertusa, using enzyme mixtures consisting of cellulase and abalone power and identified by calcofluor fluorescence. The vitality of protoplasts varied depending on the alga growth stage, and those isolated from younger tissue in March maintained a high vitality with high sealing success rate compared with protoplasts isolated from mature or non-growing plants in August or November. In the whole-cell configuration, large inward currents were elicited by negative voltage pulses. The voltage-dependent component was predominantly carried by Cl⁻, as confirmed by the use of the Cl⁻ channel inhibitor DIDS and reversal potential of current-voltage plots. This evidence suggests that hyperpolarization-activated Cl⁻ permeable channels are responsible for the influx of Cl⁻ into U. pertusa cells. Voltage-dependent outward currents were also recorded in several protoplasts, and their properties need further investigation.     

Anomalous substrate specificities among the algal peroxidases

Semipurified tissue preparations from 13 red and brown algae oxidized pyrogallol and p-coumaric acid but could not oxidize guaiacol and other ortho (methoxy)-substituted phenols, including common lignin precursors such as coniferaldehyde. They also failed to oxidize the aromatic amine, benzidine. In contrast, preparations from green algae were like horseradish peroxidase and vascular plant preparations in their ability to oxidize unsubstituted phenols, those substituted at one or both ortho-positions, and benzidine. One brown alga, Postelsia, was also unable to oxidize the commonplace peroxidase substrates, iodide and eugenol. These results suggest a phylogenetic limitation on the potential for lignification based upon enzyme stereospecificity.     

Chemical examinations of some non-vascular Paleozoic plants

The organic chemical constituents of compression fossils ofNematothallus,Spongiophyton, Orestovia andEohostimella are identified and compared with those isolated from living and fossil forms ofBotryococcus (a green alga) andTaeniocrada (a vascular plant fossil). The range and maxima in the carbon numbers observed in the normal, saturated acids isolated fromNematothallus, Orestovia, andSpongiophyton are similar to those of fossilBotryococcus, while those acids contained within compression fossils ofEohostimella are similar to the hydrocarbon composition ofTaeniocrada. Isoprenoid, branched hydrocarbons and steroids identified fromNematothallus, Orestovia, andSpongiophyton suggest these genera have algal affinities, while the presence of thick cuticles and in some cases cutin-like compounds appear to show adaptation to a terrestrial environment. Phenolic compounds retained within rock matrices associated withEohostimella are similar to those isolated fromTaeniocrada suggesting chemical, as well as morphological parallels with the land plant habit. These data are interpreted as indicating an early polyphyletic exploitation of the terrestrial habitat during the Paleozoic.     

DNA-Binding Proteins and Structural Characteristics of Chloroplast Nucleoids

A comparative analysis of proteins from chloroplast nucleoids was performed in two higher-plant species (Pisum sativumL. andArabidopsis thalianaL.) and a green alga Chlamydomonas reinhardtiiDang. In the nucleoids of the higher plants and the alga, 26–27 proteins were detected with their mol wts ranging from 10 to more than 94 kD. In all the species tested, the distribution of nucleoid proteins by their mol wts was similar, especially between the predominant proteins with mol wts of 10 to 40 kD. Six DNA-binding proteins (12–18 kD) were detected in nucleoids fromCh. reinhardtiichloroplasts after in vivocovalent cross-linking between chloroplast DNA and proteins. Under an electron miscroscope, some regular structures resembling nucleosome-like particles of bacterial cells were revealed.     

Bioactive volatile compounds from marine algae: feeding attractants

Chemical communications play an important role in plants, fungi, and algae. Volatile organic compounds in marine algae are released into the seawater. These compounds play a role as either pheromones or allelochemicals. We observed that the turbinid gastropod Lunella coronata coreensis inhabits the intertidal zone and often grazes the green alga Ulva pertusa. Feeding tests and feeding preference studies were performed with green, brown and red algae or by using the powdered freeze-dried seaweed in agar. The snails fed on U. pertusa preferentially compared to the other marine algae, and recognized chemoreception compounds from the alga but not their structural or morphological differences. From feeding tests using artificial foods, it is suggested that the feeding attractants are in the essential oil of the alga U. pertusa.     

Temperature dependence of nitrate reductase activity in marine phytoplankton: biochemical analysis and ecological implications

The temperature dependence of NADH:NR activity was examined in several marine phytoplankton species and vascular plants. These species inhabit divergent thermal environments, including the chromophytes Skeletonema costatum (12–15° C), Skeletonema tropicum (18–25° C), Thalassiosira antarctica (?2 to 4° C), and Phaeocystis antarctica (?2 to 4° C), the green alga Dunaliella tertiolecta (14–28° C), and the vascular plants Cucurbita maxima (20–35° C) and Zea mays (20–25° C). Despite the difference in growth habitats, similar temperature response curves were observed among the chromophytic phytoplankton, with temperatures optimal for NR activity being between 10–20° C. In contrast, the chlorophyll b‐containing alga and vascular plants exhibited optimal temperatures for NR activity above 30° C. Such dramatic differences in NR thermal characteristics from the two taxonomic groups reflect a divergence in NR structure that may be associated with the evolutionary diversification of chromophytes and chlorophytes. Further, it suggests a potential contribution of the thermal performance of NR to the geographic distributions, seasonal abundance patterns, and species composition of phytoplankton communities. NR partial activities, which assess the individual functions of Mo‐pterin and FAD domains, were evaluated on NR purified from S. costatum to determine the possible causes for high temperature (>20° C) inactivation of NR from chromophytes. It was found that the FAD domain and electron transport among redox centers were sensitive to elevated temperatures. S. costatum cells grown at 5, 15, and 25° C exhibited an identical optimal temperature (15° C) for NADH:NR activity, whereas the maximal NR activity and NR protein levels differed and were positively correlated with growth temperature and growth rate. These findings demonstrate that thermal acclimation of NO₃^? reduction capacity is largely at the level of NR protein expression. The consequences of these features on NO₃^? utilization are discussed.     

Molecular Evolution of Lycopene Cyclases Involved in the Formation of Carotenoids in Eukaryotic Algae

Carotenoids play crucial roles in structure and function of the photosynthetic apparatus of bacteria, algae, and higher plants. The formation of carotenoids from lycopene is catalyzed by the enzyme lycopene cyclase (LCY), which is structurally and functionally conserved in all organisms. A comparative genomic analysis regarding the LCY revealed that the higher plant (Arabidopsis thaliana) and the green alga (Ostreococcus sp. RCC809, Ostreococcus tauri, Ostreococcus lucimarinus, Micromonas sp. RCC299, Micromonas pusiua, Chlorella vulgaris, Volvox carteri, and Coccomyxa sp. C-169) possess two different LCY (beta- and epsilon-type). This indicated that an ancient gene duplication event must have occurred, which produced two classes of LCY in algae. However, some other green alga retained only one class of LCY, such as Haematococcus pluvialis (beta), Dunaliella salina (beta), Chlamydomonas reinhardtii (epsilon), and Chlorella sp. NC64A (epsilon), and the other gene copy was lost in these species. Furthermore, the similar LCY lost occurred in red alga (Cyanidioschyzon merolae) and Heterokontophyta (Phaeodactylum tricornutum and Thalassiosira pseudonana), which possess only the LCYB. In addition, the protein sequence of LCYB is highly similar to capsanthin–capsorubin synthase (CCS), which is another carotenogenic enzyme of plants. As a result, it is proposed that the CCS evolved from a duplicated LCYB. The discovery of two classes of LCY families in some algae suggests that carotenoid biosynthesis is differentially regulated in response to development and environmental stress in these algae, like members of LCY families are differentially regulated during development or stress in some higher plants.