Culture studies on Radula flaccida Lindenb. & Gottsche

Abstract

Radula flaccida has been grown on defined inorganic media. Its growth varies with the type and concentration of the media, lower concentrations proving better than higher ones. Basile's medium supported a better growth of the liverwort than that of Diller et al., Müller's medium being least effective. The liverwort was successfully established on leaves and bark of some higher plants, thus proving that it is not ‘host’ specific with respect to leaf types and that it is not exclusively epiphyllous. Its less frequent occurrence on bark may be due to other habitat factors.     

AXENIC CULTURES OF ACETABULARIA (CHLOROPHYTA): A DECONTAMINATION PROTOCOL WITH POTENTIAL APPLICATION TO OTHER ALGAE1

We developed an easy, reliable decontamination protocol for caps of Acetabularia acetabulum (L.) Silva; with minimal labor hundreds of caps can be decontaminated. In addition, cysts isolated from these caps do not exhibit dormancy and can be used immediately to establish large populations of axenic cell cultures. This method consists of three successive incubations: 1) proteinase K/sodium dodecyl sulfate/1,3-bis[tris(hydroxymethyl)-methylamino]propane for 1 h, 2) mild silver protein for 5 min, and 3) an antibiotic solution (neomycin, chloramphenicol, nystatin, ampicillin, streptomycin)for 3 days. This protocol eliminates bacterial, algal, fungal, and yeast contaminants. It is useful for decontaminating caps from lab cultures and caps collected from the wild and may also be effective in decontaminating the reproductive structures of other algae. Cyst dormancy was reduced from 15 weeks to less than 1 week, which represents a 40% reduction of the life cycle of A. acetabulum. We routinely obtained 90-100% gamete release from cysts 3-14 days after they were made axenic. The component of our culturing methods that allows gamete release without a “dormant” period is unknown.     

ANATOMICAL DEVELOPMENT OF THE LEAF TRICHILIUM AND MÜLLERIAN BODIES OF CECROPIA PELTATA L.

The anatomical development of trichilia and Müllerian bodies of Cecropia peltata was investigated at the light microscope level. The petiole base is initiated in a normal manner and the abaxial surface becomes keel-shaped during later stages of development. At maturity the trichilium is well supplied with vascular tissue but no transfer cells were observed. Before Müllerian body development, trichilium epidermal cells become modified into two types of trichomes. Müllerian bodies are initiated in sub-epidermal tissue and by continued cell division and expansion reach a size of 1 mm wide X 3 mm in length. There is no storage polysaccharide present during early stages of Müllerian body development. Glycogen production and storage begins as the food body reaches three-fourths final size and continues after cell division ceases. The mature Müllerian body is mutlicellular and rich in lipid and glycogen.     

PRODUCTION OF MALE GAMETES AND AUXOSPORES IN THE CENTRIC DIATOMS CYCLOTELLA MENEGHINIANA AND C. CRYPTICA12

Vegetative cells of Cyclotella meneghiniana Kützing form male gametes and anxospores following transfer from a synthetic freshwater medium to a modified artificial seawater. Both spermatogenesis and auxospore formation are correlated with an increase in the Na⁺ concentration in the medium. Spermatogenesis can also be induced in C. cryptica Reimann, Lewin, and Guillard in artificial seawater with an adjusted sodium level.     

EVOLUTION OF AN ARTIFICIAL SEAWATER MEDIUM: IMPROVEMENTS IN ENRICHED SEAWATER, ARTIFICIAL WATER OVER THE LAST TWO DECADES

Although most phycologists use natural seawater for culturing marine species, artificial media continue to play important roles in overcoming problems of supply and seasonal variability in the quality of natural seawater and also for experiments involving manipulation of micro- and macronutrients. Several artificial media have been developed over the last 90 years; enriched seawater, artificial water (ESAW) is among the more popular recipes. ESAW has the advantage of an ionic balance that is somewhat closer to that of normal seawater. The original paper compared the growth of 83 strains of microalgae in natural seawater (ESNW) versus ESAW and determined that 23% grew more poorly in the artificial water. Since 1980, however, the composition of ESAW, as used by the original authors, has changed considerably. In particular, the added forms of phosphate, iron, and silicate have been changed and the trace metal mixture has been altered to include nickel, molybdenum, and selenium. We tested whether these changes improved the ability of the artificial medium to grow previously difficult to grow phytoplankton species. To test this, we selected eight species that had been shown to grow better in ESNW than in ESAW and compared their growth again, using the currently used recipe with all the above modifications. For all but one species ( Apedinella spinifera ), growth rate and final yield was no different between the media but in one case ( Emiliania huxleyi ) was slightly higher in ESAW. No differences in cell morphology or volume were found in any case. We conclude that changes to the enrichment portion of the recipe have significantly improved this artificial seawater medium and that it can be used to grow an even wider range of coastal and open ocean species.     

Why are mycorrhizal fungi and symbiotic nitrogen-fixing bacteria not genetically integrated into plants?

The bacteria and fungi involved in diazotrophic and mycorrhizal symbioses with plants have varying degrees of genetic recombination. Those with very limited, or no, recombination are at risk from the progressive accumulation of mutations (‘Müller's ratchet'). This accumulation could be alleviated by complete genetic integration of the parts of the symbiont genomes relating to their function in hospice (nitrogen fixation and acquisition of a range of nutrients, respectively) into the nuclear genome of the (usually) sexually reproducing exhabitant. This integration is not known to have occurred, and the diazotrophic and mycorrhizal inhabitants with very restricted recombination survive ‘Müller's ratchet', as do some other symbioses, free-living organisms that have reproduced asexually for many generations and the organelle genomes of mitochondria and plastids. An evolutionary and environmental possibility is that it was difficult to maintain genetically integrated diazotrophs and mycorrhizas as environments changed with respect to the availability of combined nitrogen and of phosphorus. Genetic modification techniques could be used to attempt to genetically integrate diazotrophs and mycorrhizas; even granted that this could be done, a careful evaluation is required of whether the benefits of such manipulation outweigh whatever costs are entailed.     

Vittia salina L.Hedenäs & J.Muñoz,sp. nov., a new moss from Argentina

Abstract

A survey of the species of Campylopus Brid. reported from Sri Lanka (Ceylon) is presented. Of the 34 species that have been reported from this Island 16 are accepted at present. Campylopus nilghiriensis (Mitt.) Jaeg. is identical partly with C. goughii (Mitt.) Jaeg., partly with C. zollingerianus (C. Müll.) Bosch &; Lac., C. pseudogracilis Card. &; Dix. with C. goughii (Mitt.) Jaeg., C. caudatus (C. Müll.) Mont., C. reduncus (Reinw. &; Hornsch.) Bosch &; Lac. and C. trachythecius (C. Müll.) Jaeg. with C. comosus (Reinw. &; Hornsch.) Bosch &; Lac., C. herzogii Broth., C. subtricolor Lor. and probably also C. nodijlorus (C. Müll.) Jaeg. with C. aureus Bosch &; Lac., C nietneri (C. Müll.) Jaeg. with C. involutusz (C. Müll.) Jaeg., C. subgracilis Ren. &; Card. ex Gangulee and C. latinervis (Mitt.) Jaeg. with C. gracilis (Mitt.) Jaeg., and C. laetus (Mitt.) Jaeg. with C. savannarum (C. Müll.) Mitt. C. pterotoneuron (C. Müll.) Jaeg. is reduced to a variety of C. umbellatus (Arn.) Par. The occurrence of C. exasperatus Brid. on Sri Lanka could not be confirmed. Campylopus flagelliferus (C. Müll.) Jaeg. is reported as new to Sri Lanka.     

Biased predation could promote convergence yet maintain diversity within Müllerian mimicry rings of Oreina leaf beetles

Müllerian mimicry is a classic example of adaptation, yet Müller's original theory does not account for the diversity often observed in mimicry rings. Here, we aimed to assess how well classical Müllerian mimicry can account for the colour polymorphism found in chemically defended Oreina leaf beetles by using field data and laboratory assays of predator behaviour. We also evaluated the hypothesis that thermoregulation can explain diversity between Oreina mimicry rings. We found that frequencies of each colour morph were positively correlated among species, a critical prediction of Müllerian mimicry. Predators learned to associate colour with chemical defences. Learned avoidance of the green morph of one species protected green morphs of another species. Avoidance of blue morphs was completely generalized to green morphs, but surprisingly, avoidance of green morphs was less generalized to blue morphs. This asymmetrical generalization should favour green morphs: indeed, green morphs persist in blue communities, whereas blue morphs are entirely excluded from green communities. We did not find a correlation between elevation and coloration, rejecting thermoregulation as an explanation for diversity between mimicry rings. Biased predation could explain within‐community diversity in warning coloration, providing a solution to a long‐standing puzzle. We propose testable hypotheses for why asymmetric generalization occurs, and how predators maintain the predominance of blue morphs in a community, despite asymmetric generalization.     

Growth of Bacillus methanolicus in seawater-based media

Bacillus methanolicus has been proposed as a biocatalyst for the low cost production of commodity chemicals. The organism can use methanol as sole carbon and energy source, and it grows aerobically at elevated temperatures. Methanol can be made available from off-shore conversion of natural gas to methanol, through gas-to-liquid technology. Growth of the organism in seawater-based medium would further reduce the costs of chemical production performed near an off-shore natural gas source. The growth of strain PB1 (ATCC 51375) in shake flask experiments with trypticase soy broth medium showed minimal salt-inhibition at the concentration of NaCl in seawater. The ability of B. methanolicus PB1 to grow in Pacific Ocean water using methanol as a carbon and energy source was also tested. Following a simple adaptation procedure, PB1 was able to grow on methanol in semi-defined medium with 100% seawater with good growth yields and similar growth rates compared with those achieved on media prepared in deionized water.     

向日葵的幼胚培养

实验所用的基本培基中以B_5培养基对向日葵幼胚培养为最适。激素萘乙酸主要促进幼胚的胚性生长。吲哚乙酸(IAA)与激动素(KT)的配合使用,对幼胚的分化起明显的调节作用。天然提取物则明显地促进幼胚生长、其中荸荠汁的培养基可使向日葵球形胚进行正常发育。 用两种改良的B_5培养基做胚胎培养系统,使向日葵种间杂交胚生长发育。先将杂交后的心形胚培养在胚胎生长培养基上,进行充分的胚性生长。然后把胚转移到萌发培养基,使其萌发形成幼苗,最后移入土壤直至开花。     

UPTAKE OF INORGANIC CARBON BY CLADOPHORA GLOMERATA (CHLOROPHYTA) FROM THE BALTIC SEA1

Carbon uptake in the green macroalga Cladophora glomerata (L.) Kütz. from the brackish Baltic Sea was studied by recording changes in pH, alkalinity, and inorganic carbon concentration of the seawater medium during photosynthesis. The use of specific inhibitors identified three uptake mechanisms: 1) dehydration of HCO₃ ^? into CO₂ by periplasmic carbonic anhydrase, followed by diffusion of CO₂ into the cell; 2) direct uptake of HCO₃ ^? via a 4,4′‐diisothiocyanato‐stilbene‐2,2′‐disulfonate‐sensitive mechanism; and 3) uptake of inorganic carbon by the involvement of a vanadate‐sensitive P‐type H ⁺ ‐ATPase (proton pump). A decrease in the alkalinity of the seawater medium during carbon uptake, except when treated with vanadate, indicated a net uptake of the ionic species contributing to alkalinity (i.e. HCO₃ ^? , CO₃^{2 ?} , and OH ^? ) from the medium, where OH ^? influx is equivalent to H ⁺ efflux. This would suggest that the proton pump is involved in HCO₃ ^? transport. We also show that the proton pump can be induced by carbon limitation. The inducibility of carbon uptake in C. glomerata may partly explain why this species is so successful in the upper littoral zone of the Baltic Sea. Usually, carbon limitation is not a problem in the upper littoral of the sea. However, it may occur frequently within dense Cladophora belts with high photosynthetic rates that create high pH and low carbon concentrations in the alga's microenvironment.     

Distribution maps of Bryophytes in Britain and Ireland

Abstract

Eight epithets of Rigodium (R. araucarieti C. Müll., R. argentinicum C. Müll., R. brachypodium (C. Müll.) Par., R. nano-fasciculatum C. Müll. ex Thér., R. pendulum Herz. &; Thér., R. pseudo-thuidium Dus., R. solutum (Tayl.) Par., and R. tamarix C. Müll), lectotypified here, are listed with their nomenclature and pertinent details involving the careful identification of type specimens and original material. For Rigodium solutum (Tayl.) Par., a specimen from Taylor's original herbarium may likely be the holotype but had not been indicated as such by Taylor, the original author. A conservative approach for this situation is taken here by designating this specimen as a lectotype.     

On the identities of some neglected species of Grimmiaceae from Îles Kerguelen

Taxonomic status of five species of Grimmia Hedw. (G. minuta Müll.Hal., G. suborthotrichacea Müll.Hal., G. defoliata Müll.Hal., G. genuflexa Müll.Hal., and G. orthotrichacea Müll.Hal.) is appraised. All these species correctly belong within the genus Bucklandiella Roiv. They are actually inseparable from one another and represent a single species for which B. orthotrichacea (Müll.Hal.) Bednarek-Ochyra & Ochyra is chosen as the final name. The remaining four names are considered its synonyms, all but G. genuflexa being new. All the names are lectotypified and some details of the leaves of G. minuta and G. defoliata are illustrated.     

Mosses of southwest Ethiopian montane forests – notes on their occurrence pattern and many new country records

One hundred and thirty-nine taxa of mosses are reported from the montane forest zone of southwest Ethiopia. Of these, 53 taxa are new country records and Meteoriopsis reclinata (Müll.Hal.) M.Fleisch. ex Broth. is, in addition, new to Africa. Most of the new records are of species also found in Uganda, Kenya or Tanzania, but there are some surprises, such as Fabronia perciliata Müll.Hal. with the closest location in South Africa and Namibia, Entodontella cameruniae Broth. previously known only from West Africa and Bryomaltaea obtusifolia (Hook.) Goffinet, otherwise reported only from the Democratic Republic of the Congo. We list all taxa and give a short account of their frequency and in which habitat and on which substrate they grow. That such a large proportion of the species are new to the country suggests that Ethiopia is bryologically very under-explored, and that further exploration will reveal many additional species. In particular, the remnant forests of southwest Ethiopia are likely to be of great importance as bryophyte hotspots.     

INFLUENCE OF SALINITY ON THE FORM OF ASTEROCYTIS IN PURE CULTURE1

The growth form of an isolate of Asterocytis ornata in pure culture changed from filamentous in a full-strength seawater medium to a unicellular or bicellular form closely resembling the genus Chroothece in a medium of quarter-strength seawater. The alga did not grow in media of lower salinities, or in media in which seawater was replaced by KCl, NaCl, or CaCl₂. The formation of filaments from single cells could not be attributed to osmolarity. The controlling factors seemed to be the concentrations of cations, notably Na⁺ and Mg⁺⁺. However, the filaments produced experimentally in a defined medium, were never as extensively developed as those occurring in seawater, an indication that further studies should be done. The occurrence of the Asterocytis form, in freshwater habitats remains to be explained. There may be distinct physiological races, if not true species, within the genus.     

Comparing the low‐salinity tolerance of Ulva species distributed in different environments

The green macroalgal genus Ulva (Ulvales, Ulvophyceae, Chlorophyta) is distributed worldwide from marine to freshwater environments. Comparative analyses of hyposalinity tolerance among marine, brackish, and freshwater Ulva species were performed by fluorescein diacetate viability counts. The subtidal marine species Ulva sp., collected from a depth of 30 m, showed the poorest tolerance to low salinity. This species died in 5 practical salinity units (PSU) artificial seawater or freshwater within 1 day. Its closely related species U. linza L. (an intertidal species) and U. prolifera Müller (a brackish species) showed varying tolerances to low salinity. After 7 days of freshwater exposure, the viability of U. linza L. decreased to approximately 20%, while U. prolifera Müller showed nearly 100% viability. The freshwater species U. limnetica Ichihara et Shimada, not yet found in coastal areas, was highly viable in seawater.     

Effects of Freeze-Drying on Permeability and Respiration of Germinating Lily Pollen

The germination of lily pollen (Lilium longiflorum cv. Ace) was impaired by freeze-drying. This loss of viability was associated with a modified pattern of respiration and an increased leakage of soluble carbohydrates, phosphate, and ninhydrin-positive material into the culture medium. 2,4-Dinitro-phenol, (DNP), an uncoupler of oxidative phosphorylation showed a decreased ability to stimulate O₂ uptake in freeze-dried pollen. The altered viability, respiration, and permeability resulted from drying under vacuum and not the initial freezing of the pollen. Mature lily pollen contained approximately 0.3 “Ai phosphate, of which 15 % was inorganic phosphate and about 50 %> was acid soluble organic phosphate of unknown identity.     

A revision of the moss genus Gymnostomiella Fleisch.

Abstract

A taxonomic revision of the moss genus Gymnostomiella Fleisch. recognizes five species and one variety, viz. G. burmensis Bartr., G. erosulum (C. Müll. ex Dus.) Arts, G. longinervis Broth., G. monodii P. Yarde, G. vernicosa (Harv.) Fleisch. var. vernicosa and G. vernicosa var. tenerum (C. Müll. ex Dus.) Arts. Gymnostomiella tenerum (C. Müll. ex Dus.) Arts is reduced to a variety of G. vernicosa and G. tanganyikae De Sloover is reduced to synonomy with G. erosulum.     

Tasting the difference: do multiple defence chemicals interact in Müllerian mimicry?

Müllerian mimicry, where two unpalatable species share a warning pattern, is classically believed to be a form of mutualism, where the species involved share the cost of predator education. The evolutionary dynamics of Müllerian mimicry have recently become a controversial subject, after mathematical models have shown that if minor alterations are made to assumptions about the way in which predators learn and forget about unpalatable prey, this textbook case of mutualism may not be mutualistic at all. An underlying assumption of these models is that Müllerian mimics possess the same defence chemical. However, some Müllerian mimics are known to possess different defence chemicals. Using domestic chicks as predators and coloured crumbs flavoured with either the same or different unpalatable chemicals as prey, we provide evidence that two defence chemicals can interact to enhance predator learning and memory. This indicates that Müllerian mimics that possess different defence chemicals are better protected than those that share a single defence chemical. These data provide insight into how multiple defence chemicals are perceived by birds,and how they influence the way birds learn and remember warningly coloured prey. They highlight the importance of considering how different toxins in mimicry rings can interact in the evolution and maintenance of Müllerian mimicry and could help to explain the remarkable variation in chemical defences found within and between species.     

Diversity in Müllerian mimicry: The optimal predator sampling strategy explains both local and regional polymorphism in prey

The convergent evolution of warning signals in unpalatable species, known as Müllerian mimicry, has been observed in a wide variety of taxonomic groups. This form of mimicry is generally thought to have arisen as a consequence of local frequency‐dependent selection imposed by sampling predators. However, despite clear evidence for local selection against rare warning signals, there appears an almost embarrassing amount of polymorphism in natural warning colors, both within and among populations. Because the model of predator cognition widely invoked to explain Müllerian mimicry (Müller's “fixed ” model) is highly simplified and has not been empirically supported; here, we explore the dynamical consequences of the optimal strategy for sampling unfamiliar prey. This strategy, based on a classical exploration–exploitation trade‐off, not only allows for a variable number of prey sampled, but also accounts for predator neophobia under some conditions. In contrast to Müller's “fixed ” sampling rule, the optimal sampling strategy is capable of generating a variety of dynamical outcomes, including mimicry but also regional and local polymorphism. Moreover, the heterogeneity of predator behavior across space and time that a more nuanced foraging strategy allows, can even further facilitate the emergence of both local and regional polymorphism in prey warning color.