Physiological and biochemical contributions to the taxonomy of the genera Ankistrodesmus and Scenedesmus

The 17 Ankistrodesmus strains studied do not produce extracellular amylolytic enzymes. 20 out of 28 Scenedesmus strains, however, are able to hydrolyse starch.The groups of strains previously characterized physiologically and biochemically were assigned to established species of the genera Ankistrodesmus (including Monoraphidium) and Scenedesmus.Abbreviation GC Guanine + cytosine     

Über das ätherische Öl von Grünalgen

Zusammenfassung Das ätherische Öl und seine Hauptkomponenten (Proazulene) erwiesen sich auch bei den Gattungen Ankistrodesmus und Scenedesmus als brauchbare taxonomische Merkmale. Der Ölgehalt licgt in der gleichen Größenordnung wie innerhalb der Gattung Chlorella. Einige Ankistrodesmus-Stämme, die keine Proazulene bilden, müssen offensichtlich anderen Gattungen zugeordnet werden.

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On the essential oil of green algaeII. The oils of the genera Ankistrodesmus and Scenedesmus

The essential oil and its main components (i.e., proazulenes) are useful taxonomic characters also in the genera Ankistrodesmus and Scenedesmus. The amounts of oil in these genera are similar to that of the genus Chlorella. A few strains of Ankistrodesmus, which are unable to synthesize proazulenes, seem to belong to other genera.

     

Growth of Corbicula fluminea (bivalvia) fed artificial and algal diets

Laboratory experiments were conducted to assess the growth of Corbicula fluminea Müller (1774) fed mixed diets of the green alga Ankistrodesmus and five artificial foods (nine-grain cereal, rice flour, rye bran, denatured brewer's yeast and Ralston Purina trout chow). Clams lost weight after 30 days on each artificial diet; however, weight losses were significantly less than those of control clams which were not fed (p < 0.05). In contrast, clam growth was positive in each treatment which included Ankistrodesmus. There was no significant difference in the tissue growth of clams fed on an Ankistrodesmus monoculture and Ankistrodesmus supplemented with each artificial diet (p > 0.2). Therefore, we conclude that all of the artificial diets evaluated were of nutritional value. However, the alga Ankistrodesmus appears to be a superior food item. In subsequent experiments, clams were fed on diets consisting of various combinations of six genera of green algae (Pedinomonas, Ankistrodesmus, Chlamydomonas, Chlorella, Scenedesmus, and Selenastrum). All trialgal combinations of Chlamydomonas, Chlorella, Scenedesmus, and Ankistrodesmus and a dialgal culture of Chlamydomonas and Chlorella resulted in strong positive tissue growth. No other dialgal or monoalgal culture resulted in similar growth. Cultures of Selenastrum were found to be toxic, and their filtrate was determined to inhibit Corbicula filtration.     

Physiologische und biochemische Beiträge zur Taxonomie der Gattungen Ankistrodesmus und Scenedesmus

The base composition of the DNA was found to be a species-specific, taxonomically valuable character also in the genera Ankistrodesmus (21 strains) and Scenedesmus (27 strains). As compared to the genus Chlorella, however, the differences between different species are considerably smaller. The GC content is between 63 and 70% in the genus Ankistrodesmus, and between 52 and 62% in the genus Scenedesmus. Obviously, a few strains with a very much different base composition do not belong to the genus Ankistrodesmus.     

Algal bioassays to determine toxicity of metal mixtures

The shortcomings of many established toxicity criteria for metals have resulted from a simplification of bioassays performed with single metals. A more comprehensive approach was needed to diagnose the effects of metal mixtures on aquatic organisms. Using Chlorella, Ankistrodesmus and Scenedesmus as test organisms, we experimented on a number of factors which affected metal toxicity bioassays. These included metal interactions, algal competitions, species sensitivities, the ratio of an excess metal to other metals and nutrient levels. An alternative technique was finally established which involved an evaluation of individual assays of Chlorella and Ankistrodesmus in separate media and a tissue-metal analysis on Chlorella. Chlorella fusca, commonly found in lakes with high metal concentrations, showed high tolerance to mixed-metal solutions, while Ankistrodesmus proved to be very sensitive. By determining the maximum yield ratio between Ankistrodesmus and Chlorella (i.e. the A/Ch ratio) it was possible to compare the toxic strength of harmful metals according to an established standard curve of A/Ch ratio versus mixed-metal concentrations. The levels of tissue-metal analysed in Chlorella also gave some indication as to which metals were responsible for the toxicity.     

Physiologische und biochemische Beiträge zur Taxonomie der Gattungen Ankistrodesmus und Scenedesmus

Zusammenfassung Die Arten der Gattungen Ankistrodesmus und Scenedesmus, die sich hinsichtlich Hydrogenase-Aktivität, Bildung von Sekundär-Carotinoiden und Gelatine-Verflüssigung als biochemisch sehr einheitlich erwiesen hatten, zeigen zum Teil beträchtliche Unterschiede in ihrer Säureresistenz (vgl. Tab. 1–4), die somit als taxonomisches Merkmal verwendet werden kann.

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Physiological and biochemical contributions to the taxonomy of the genera Ankistrodesmus and Scenedesmus II. Acid tolerance

Summary Some species of the genera Ankistrodesmus and Scendesmus exhibit pronounced differences in acid tolerance (cf. Tables 1–4), in contrast to their biochemical uniformity in hydrogenase activity, formation of secondary carotenoids, and liquefaction of gelatin. Thus acid tolerance can serve as a taxonomic character in these genera.

     

Different tolerances and responses to low temperature and darkness between waterbloom forming cyanobacterium <Emphasis Type="Italic">Microcystis</Emphasis> and a green alga <Emphasis Type="Italic">Scenedesmus</Emphasis>

The dynamics of planktonic cyanobacteria in eutrophicated freshwaters play an important role in formation of annual summer blooms, yet overwintering mechanisms of these water bloom forming cyanobacteria remain unknown. The responses to darkness and low temperature of three strains (unicellular Microcystis aeruginosa FACHB-905, colonial M. aeruginosa FACHB-938, and a green alga Scenedesmus quadricauda FACHB-45) were investigated in the present study. After a 30-day incubation under darkness and low temperature, cell morphology, cell numbers, chlorophyll a, photosynthetic activity (ETR_max and I _k), and malodialdehyde (MDA) content exhibited significant changes in Scenedesmus. In contrast, Microcystis aeruginosa cells did not change markedly in morphology, chlorophyll a, photosynthetic activity, and MDA content. The stress caused by low temperature and darkness resulted in an increase of the antioxidative enzyme-catalase (CAT) in all three strains. When the three strains re-grew under routine cultivated condition subjected to darkness and low temperature, specific growth rate of Scenedesmus was lower than that of Microcystis. Flow cytometry (FCM) examination indicated that two distinct types of metabolic response to darkness and low temperature existed in the three strains. The results from the present study reveal that the cyanobacterium Microcystis, especially colonial Microcystis, has greater endurance and adaptation ability to the stress of darkness and low temperature than the green alga Scenedesmus. Handling editor: D. Hamilton     

Comparison of the roles of Ostracods and Cladocerans in regulating community structure and metabolism in freshwater microcosms

Laboratory microcosms were used to compare the effects of the littoral ostracod Cypridopsis vidua and the planktonic cladoceran Daphnia magna on community structure and metabolism. Filter-feeding by cladocerans, both in the presence and absence of ostracods, greatly reduced the abundance of planktonic algae when D. magna reached peak density around day 50; rotifers and euglenids were then limited to flocculent matter on the container bottom. Both net production and community respiration rates decreased as community composition changed. Microcosms containing ostracods as the only microcrustacean showed little reduction in total algal numbers but the otherwise dominant alga, Scenedesmus spp., was replaced by Ankistrodesmus spp. when peak ostracod density was reached around day 100. Rotifers were completely eliminated but euglenids were able to coexist with ostracods. Ostracods impacted community metabolism less than cladocerans, but depressed respiration slightly more than net production.     

Survival of Chlorophyceae Ingested by Saprozoic Nematodes

The saprozoic nematode, Pristionchus lheritieri ingested cells of four species of unicellular Chlorophyceae (grass-green algae) including Chlamydomonas reinhardi and unidentified species of Ankistrodesmus, Chlamydornonas and Scenedesmus. Additional tests with Ankistrodesmus sp. and Chlamydomonas sp., indicated cells of Ankistrodesmus survived passage through the alimentary canal and were subsequently cultured, while viable cells of Chlarnydomonas were only occasionally recovered.     

Importance of Nutrient Competition and Allelopathic Effects in Suppression of the Green Alga Scenedesmus obliquus by the Macrophytes Chara, Elodea and Myriophyllum

Possible allelopathic effects of substances released from the macrophytes Chara globularis, Elodea canadensis, Myriophyllum spicatum on the common green alga Scenedesmus obliquus were tested in the laboratory with plastic plants and untreated medium as controls. A two-phase approach was used in which first the effects of physical presence of plants was studied (phase I) followed by the effects of plant culture filtrates (phase II). In the presence of plastic plants growth was reduced only marginally, but strong growth inhibition of Scenedesmus occurred in the physical presence of all macrophytes. In contrast, filtrates from Chara had no growth inhibitory effect on Scenedesmus. Myriophyllum filtrate reduced particle-based growth rate by 7% compared to filtration controls, while Elodea culture filtrate reduced volume-based growth by 12%, chlorophyll-based growth by 28% and particle-based growth by 15%. Photosystem II-efficiency of Scenedesmus was reduced in all three macrophyte treatments in phase I, but not in filtrates from macrophyte cultures (phase II). Thus, while enzyme activity or other physiological aspects may have been affected, the current study yielded no proof for allelopathically active compounds being directed at photosynthesis. Mean particle volume (MPV) of Scenedesmus was not influenced by macrophyte exudates and cultures remained dominated by unicells. The strong growth inhibitory effects found for Scenedesmus in the physical presence of macrophytes, but not in plastic controls, and no or weaker response in nutrient-enriched filtrates, suggest nutrient competition was a more powerful driving factor than allelochemicals. However, the experimental design does not exclude disappearance of allelochemicals during the filtration process.     

Kluyveromyces lactis and Saccharomyces cerevisiae, two potent deacidifying and volatile-sulphur-aroma-producing microorganisms of the cheese ecosystem

Cheese flavour is the result of complex biochemical transformations attributed to bacteria and yeasts grown on the curd of smear-ripened cheeses. Volatile sulphur compounds (VSCs) are responsible for the characteristic aromatic notes of several cheeses. In the present study, we have assessed the ability of Kluyveromyces lactis, Kluyveromyces marxianus and Saccharomyces cerevisiae strains, which are frequently isolated from smear-ripened cheeses, to grow and deacidify a cheese medium and generate VSCs resulting from l-methionine degradation. The Kluyveromyces strains produced a wider variety and higher amounts of VSCs than the S. cerevisiae ones. We have shown that the pathway is likely to be proceeding differently in these two yeast genera. The VSCs are mainly generated through the degradation of 4-methylthio-oxobutyric acid in the Kluyveromyces strains, in contrast to the S. cerevisiae ones which have higher l-methionine demethiolating activity, resulting in a direct conversion of l-methionine to methanethiol. The deacidification activity which is of major importance in the early stages of cheese-ripening was also compared in S. cerevisiae and Kluyveromyces strains.     

Phylogenetic position of someChlorella species within the chlorococcales based upon complete small-subunit ribosomal RNA sequences

Summary Complete small-subunit rRNA (16S-like rRNA) coding region sequences were determined for eight species of the Chlorococcales (Chlorophyceae). The genera investigated includePrototheca, Ankistrodesmus, Scenedesmus, and fiveChlorella species. Distance matrix methods were used to infer a phylogenetic tree that describes evolutionary relationships between several plant and green algal groups. The tree exhibits a bifurcation within the Chlorococcales consistent with the division into Oocystaceae and Scenedesmaceae, but three of the fiveChlorella species are more similar to other algae than toChlorella vulgaris. All of the sequences contain primary and secondary structural features that are characteristic of 16S-like rRNAs of chlorophytes and higher plants.Anikstrodesmus stipitatus, however, contains a 394-bp group I intervening sequence in its 16S-like rRNA coding region.     

Culturable Actinobacteria from the Marine Sponge Hymeniacidon perleve: Isolation and Phylogenetic Diversity by 16S rRNA gene-RFLP Analysis

A total of 106 actinobacteria associated with the marine sponge Hymeniacidon perleve collected from the Yellow Sea, China were isolated using eight different media. The number of species and genera of actinobacteria recovered from the different media varied significantly, underlining the importance of optimizing the isolation conditions. The phylogenetic diversity of the actinobacteria isolates was assessed using 16S rRNA gene amplification–restriction fragment length polymorphism (RFLP) analysis of the 106 strains with different morphologies. The RFLP fingerprinting of selected strains by HhaI-digestion of the 16S rRNA genes resulted in 11 different patterns. The HhaI-RFLP analysis gave good resolution for the identification of the actinobacteria isolates at the genus level. A phylogenetic analysis using 16S rRNA gene sequences revealed that the isolates belonged to seven genera of culturable actinobacteria including Actinoalloteichus, Micromonospora, Nocardia, Nocardiopsis, Pseudonocardia, Rhodococcus, and Streptomyces. The dominant genus was Streptomyces, which represented 74% of the isolates. Three of the strains identified are candidates for new species.     

Electrophoretic protein patterns of Geotrichum and its teleomorphs

Total patterns of water-soluble proteins of 35 strains (7 species) of Galactomyces and Dipodascus strains with their respective Geotrichum anamorphs are compared. Quantitative differences among a number of species are found with iso-electric focusing; bands that characterize species are selected. Qualitative differences are found between the teleomorph genera.     

Activities of superoxide dismutase (SOD) isoforms during growth of <Emphasis Type="Italic">Scenedesmus</Emphasis> (chlorophyta) species and strains grown in batch-cultures

Growth of Scenedesmus species and strains, grown for 28 days in mineral BBM medium in batch-cultures, displayed sigmoidal kinetics that comprised a lag, exponential and declining growth phases. Total SOD activity in these autotrophically cultured organisms, which oscillated within 0.6 – 1.4 Umg protein⁻¹, was rather species-specific and only to some extent depended on the growth phase. Contrary, three S. obliquus strains: wild type 276-6, mutant with blocked PS I (strain 56.80) and mutant with blocked PS II (strain 57.80), cultured for 7 days on BBM medium supplemented with bacto-tryptone and yeast extract (BBM+) turned out to be time-dependent and to have several times higher total SOD activity than one obtained for Scenedesmus grown autotrophically. Regardless of the media composition, the phase of growth and studied organism, dominant isoforms of total SOD were together determined Fe- and Mn-SOD. Profiles of SOD isoforms, obtained after PAGE analysis of all autotrophically and exponentially growing organisms, revealed that one Mn-SOD and one Cu/Zn-SOD bands located on gels at the same position whereas location of three bands of Fe-SOD depended on the strain. This suggests the presence of two different groups of Fe-SODs in analyzed organisms. Identical SOD profiles found in two S. armatus strains (276-4a and 276-4d) and S. subspicatus correspond well with their taxonomic position. The SOD profile of S. armatus B1-76 distinctly differed from two other S. armatus strains but was identical to S. microspinal B1-76 and S. quadricauda G-15 despite the fact that there were significant growth rate differences between these three species. SODs profiles of S. acutus 437 and S. obliguus 453 were species-specific. In S. obliquus strains cultured on BBM+ medium, there are four SOD bands: one slightly visible band of Mn-SOD, two intensive bands of Fe-SOD and one band of Cu/Zn-SOD. The above finding suggests that antioxidant response of algae kept in batch-cultures differs according to medium composition and the SOD activity mainly restricted to chloroplasts.     

Food quality effects on life history traits and fitness in the generalist herbivore Daphnia

Summary Life table experiments were conducted on the generalist suspension feeder Daphnia galeata, using as food the two green algae (Chlorophyta) Scenedesmus acutus and Oocystis lacustris. Oocystis was hypothesized to be a lower quality food because it is convered with a thick sheath, believed to reduce digestibility. Results showed that Oocystis is a lower quality food for Daphnia, but only at relatively low food concentrations (0.15 mg C/L) and not at higher concentrations (1.0 mg C/L). At 0.15 mg C/L, Daphnia intrinsic rate of increase (r) when grown on Oocystis was only half that when grown on Scenedesmus. Daphnia r was similar at 0.15 mg C/L Oocystis and 0.075 mg C/L Scenedesmus, indicating that Daphnia requires twice as much Oocystis as Scenedesmus to achieve the same fitness. Intrinsic rate of increase was lower on Oocystis mainly because age at first reproduction was greatly delayed compared to that on Scenedesmus (13.6 vs 7.3 d). In addition, juvenile growth and survivorship were reduced on Oocystis compared with Scenedesmus. Clutch sizes were similar on the two foods, indicating that once individuals reached adulthood, the two foods were similar in quality. In contrast, at high food concentrations (1.0 mg C/L), the two algae were similar in quality for both juveniles and adults, and r was not significantly different on the two foods. Ingestion and assimilation rate experiments whowed that Daphnia consumes the two algae at identical rates, and that adults assimilate the two algae at similar rates. However, juveniles assimilate Oocystis at much lower rates than Scenedesmus, possibly accounting for reduced juvenile growth and delay in age at maturity at low concentrations. Thus, Daphnia exhibits an ontogenetic shift in its ability to utilize Oocystis, and this can result in juvenile bottlenecks in which survival and growth of young age classes are of critical importance in determining population dynamics. Because food quality effects were manifested primarily in juveniles and at low concentrations, food quality effects in nature will depend on phytoplankton abundance and age-structure of Daphnia populations.     

高寒特境中甸黄芪植物根际微生物物种多样性及其抗生物膜活性成分

滇西北高寒地区分布着丰富的黄芪属植物资源，该属植物“根际效应”明显，其根际微生物极具抗菌药用资源研究价值。【目的】认知滇西北高寒特境中甸黄芪根际微生物的物种多样性，探究其可培养菌株次生代谢产物的化学多样性及抗菌、抗生物膜活性。【方法】采用宏基因组和微生物纯培养方法对中甸黄芪植物根际微生物进行物种多样性分析，同时采用高效液相色谱（high performance liquid chromatography，HPLC）、超高效液相色谱-质谱联用法（ultra-performance liquid chromatography-mass spectrometry，UPLC-MS）结合“微量肉汤稀释法” “孔板法”等多级联合筛选策略综合评估可培养菌株的抗菌活性药源研究价值。【结果】对中甸黄芪根际土壤样本的微生物分类操作单元（operational taxonomic units，OTU）进行分类注释，得到22门54纲105目187科316属856种微生物，其中优势菌群为慢生根瘤菌属。纯培养共获得27属54种95株可培养菌株，包括20属33种54株细菌和7属21种41株真菌，优势属分别为芽孢杆菌属和青霉属。其中，1株细菌Pseudomonas tolaasii ZTB4和3株真菌Aspergillus tabacinus ZNF17、Lecanicillium aphanocladii ZNF15、Umbelopsis nana ZTF31的次生代谢产物具有广谱抗菌活性。同时，菌株ZTB4和ZNF17的次生代谢产物也显示出优秀的抗耐甲氧西林金黄色葡萄球菌（methicillin-resistant Staphylococcus aureus，MRSA）生物膜活性，并已验证这2株菌株的主要活性成分分别为环脂肽类与黄酮类。【结论】中甸黄芪植物根际微生物物种多样性较为丰富，其可培养菌株次生代谢产物有较好的化学多样性和抗菌、抗生物膜活性。研究结果为我国特境特色微生物药用资源的开发利用提供理论依据。     

Genetic diversity and distribution of Bradyrhizobium and Azorhizobium strains associated with the herb legume Zornia glochidiata sampled from across Senegal

Herb legumes have great potential for rehabilitation of semi-arid degraded soils in Sahelian ecosystems as they establish mutualistic symbiosis with N₂-fixing rhizobia. A phylogenetic analysis was performed for 78 root nodule bacteria associated with the common Sahelian herb legume Zornia glochidiata Reichb ex DC in Senegal. Based on ITS (rDNA16S-23S) and recA sequences, these strains were shown to belong to the two genera Bradyrhizobium and Azorhizobium. Strains of this latter, although frequent, formed small and ineffective nodules and suggested a parasitism rather than a symbiotic association. A potential negative effect of Azorhizobium on Zornia growth was tested for when inoculated alone or in association with a Bradyrhizobium strain. Bradyrhizobium isolates were distributed in four groups. Groups A and B were two sister clades in a larger monophyletic group also including Bradyrhizobium liaoningense, Bradyrhizobium yuanmingense, and Bradyrhizobium japonicum. Strains of cluster D fell in a sister clade of the photosynthetic Bradyrhizobium sp. group, including ORS278, whereas group C appeared to be divergent from all known Bradyrhizobium clusters. Amplified fragment length polymorphism (AFLP) clustering was congruent with ITS and recA phylogenies, but displayed much more variability. However, within the main Bradyrhizobium clades, no obvious relationship could be detected between clustering and geographical origin of the strains. Each sub-cluster included strains sampled from different locations. Conversely, Azorhizobium strains showed a tendency in the phylogeny to group together according to the site of sampling. The predominance of ineffective Azorhizobium strains in the nodules of Zornia roots, the large Bradyrhizobium genetic diversity and the geographical genetic diversity pattern are explored.     

Antibiotic Susceptibility as a Taxonomic Characteristic of Proteobacteria of the Genera Alteromonas, Pseudoalteromonas, Marinomonas, and Marinobacter

Type strains of bacteria of the genera Alteromonas, Pseudoalteromonas, Marinomonas, and Marinobacterand symbiotrophic bacteria isolated from the Far Eastern bivalve mollusks Crenomytilus grayanusand Mizuhopecten yessoensisand identified as Pseudoalteromonas nigrifaciensand P. citreawere tested for susceptibility to antibiotics. Of the ten antibiotics used in this study, benzylpenicillin, polymyxin, lincomycin, and oleandomycin were valuable for the polyphasic taxonomy of marine aerobic proteobacteria. Marine bacteria of the genera studied exhibited characteristic antibiotic susceptibility profiles, which were useful for the differentiation of the genera.     

Grazer-induced changes in the desmid Staurastrum

In aquatic environments, predator kairomones have been shown to affect morphology of prey species. Past work on the interaction between zooplankton and phytoplankton was based mainly on the Daphnia–Scenedesmus model. Algae of the genus Staurastrum can produce mucilage, causing cell clumping and settling out of the water column. These clumps are too large to be eaten by daphniids. Thus, we hypothesised that this may be a grazer defence. We investigated whether Daphnia magna induces the formation of mucus globules in Staurastrum, how this occurs, and if the formation of clumps inhibits growth in juvenile Daphnia. Eight strains of Staurastrum were used to check whether mucus extrusion is induced by the presence of Daphniaor possibly by a chemical excreted by Daphnia magna. None of the strains reacted to the presence of Daphnia water alone, animals had to be present to induce clumping. Mechanical action (gentle stirring) caused the same strains to clump. The ecological relevance of clumping was then investigated. The different Staurastrum strains were used as food in a growth experiment with ecologically relevant densities of neonates of Daphnia hyalina. These small daphniids did not cause the same clumping observed for Daphnia magna when present in experiments at high densities. We observed that juvenile daphniids grew less well on strains with larger cell size.