CELLULAR SLIME MOLDS OF SOUTHEAST ASIA. I. DESCRIPTION OF NEW SPECIES

Sixteen species of cellular slime molds were isolated from Southeast Asian forest soils. Ten of these, Dictyostelium mucoroides Brefeld, D. purpureum Olive, D. polycephalum Raper, D. lacteum van Tieghem, D. rhizopodium Raper and Fennell, D. lavandulum Raper and Fennell, D. vinaceo-fuscum Raper and Fennell, D. coeruleo-stipes Raper and Fennell, Polysphondylium violaceum Brefeld, and P. pallidum Olive have been previously described and are well-recognized species occurring in other parts of the world. Two, one in the genus Dictyostelium and one belonging to the family Guttulinaceae, are considered species by the author but have not been formally described. Four are described in this paper as new: Dictyostelium intermedium, D. multi-stipes, D. bifurcatum, and Acytostelium subglobosum. A new variety papilloideum of D. lacteum is also described. One other discovery of special interest is an isolate of Polysphondylium violaceum which produces abundant macrocysts, now known to be the sexual stage in the life cycle of cellular slime molds.     

THE YELLOW-PIGMENTED DICTYOSTELIA

The cellular slime molds described herein are of a clear golden yellow color and belong to two naturally occurring groups. One we call Dictyostelium mexicanum sp. n. because it is found in soils from various parts of tropical and subtropical Mexico. The well-tapered stalk and disklike base are somewhat reminiscent of D. discoideum. The other is regarded as D. aureum E. W. Olive (1901) with which we feel it is closely identified if not identical. In the latter group the form of both the aggregation and sorocarp is suggestive of D. mucoroides. Some less pigmented isolates are separated as D. aureum var. luteolum var. n.     

Description of dictyostelid cellular slime mold species in Norway

Ten different species of dictyostelid cellular slime molds were found in Norway, namely Dictyostelium mucoroides Brefeld, D. sp. 1, D. sp. 2, D. aureo–stipes Cavender, Raper & Quinlan, D. fasckulatum Traub, Hohl & Cavender, D. sp. 3, D. minutum Raper, Polysphondylium violaceum Brefeld, P. pallidum Olive and Acylo–stelium leptosomum Raper & Quinlan. D. mucoroides included two varieties, D. sp. 1 and D. sp. 2 are probably related to D. septentrionalis Cavender and D. sphaerocephalum (Oud.) Sacc. & March respectively and D. sp. 3 is probably a new species.
Two different spore types, characterized by the presence or absence of ribosomes on the outer mitochondrial membranes, were revealed by transmission electron microscopic studies. This property was related to other properties such as the presence or absence of polar spore granules (PG) and the aggregative responses of the amoebae towards cAMP.     

CELLULAR SLIME MOLDS OF SWITZERLAND. I. DESCRIPTION OF NEW SPECIES

During the study of the distribution of cellular slime molds in Swiss forest soils, three new species were isolated and studied in culture, namely, Dictyostelium fasciculatum, Dictyostelium polycarpum and Polysphondylium filamentosum. The first is cosmopolitan, the latter two are known only from Switzerland. Dictyostelium polycarpum is a very delicate species and is the first species reported which is restricted to a high altitude environment. It is characterized by a coremiform habit, reminiscent of D. polycephalum, and relatively long reniform spores. Dictyostelium fasciculatum is a larger species which resembles D. mucoroides in gross morphology but is much closer to Polysphondylium in its life cycle pattern and behavior. Polysphondylium filamentosum is only the third well-defined species of the genus to be described. It produces filamentose principal and lateral axes. Optimum temperature for all three is around 20 C, somewhat below that of most other species in the family.     

A NATURALLY OCCURRING DEVELOPMENTAL SYNERGISM BETWEEN THE CELLULAR SLIME MOLD,DICTYOSTELIUM MUCOROIDES AND THE FUNGUS,MUCOR HIEMALIS

We here report the second record of a developmentally aberrant strain of a cellular slime mold from natural populations and demonstrate that this Dictyostelium mucoroides variant is capable of undergoing normal morphogenesis in the presence of the phycomycete fungus, Mucor hiemalis. The synergism is induced by an extracellular product(s) which is diffusable through thin agar membranes and is released by the fungus. The presence of the fungus not only induces stalk formation in this stalkless variant, but also increases the rate of sorocarp formation in 3 of 5 additional species of cellular slime molds assayed.     

Distinct lectin activities from six species of cellular slime molds.

Extracts of cohesive cells of four species of cellular slime mold, D. mucoroides, D. purpureum, D. rosarium and P. violaceum agglutinate erythrocytes in a manner that is similar to that previously observed with extracts of D. discoideum and P. pallidum. We determined inhibitory activity of a series of sugars on the agglutination activity of each of these extracts, using both semiquantitative and quantitative agglutination assays. The inhibitory potency of this series of sugars was distinct for each extract, although only slight differences were found between several species, especially D. discoideum and P. violaceum. A possible role of these agglutinins in species-specific cell cohesion is considered.     

4D confocal microscopy of Dictyostelium discoideum morphogenesis and its presentation on the Internet

 Methods to present three-dimensional (3D) and time series of 3D datasets (4D) are demonstrated using the recent advances in confocal microscopy and computer visualization. The process of cell sorting during tip formation in the slime mould Dictyostelium discoideum is examined as an example by in vivo confocal microscopy of spectrally different green fluorescent protein (GFP) variants as reporters of cell-type specific gene expression. Also, cell sorting of the co-aggregating slime mould species D. discoideum and D. mucoroides is observed using a GFP variant and a spectrally distinguishable fluorescent vital stain. The confocal data are handled as 3D and 4D datasets, their processing and the advantages of different methods of visualization are discussed step by step. Selected sequences of the experiments can be viewed on the Internet, giving a much better impression of the complex cellular movements during Dictyostelium morphogenesis than printed photographs. Received: 17 February 1998 / Accepted: 14 June 1998     

Dictyostelids from Mediterranean forests of the south of Europe

The first results of a study of dictyostelid diversity in soils of Mediterranean vegetation are presented. Surface soil samples were collected during 2003–2004 from different Mediterranean forests in Spain and Portugal and were plated out for cellular slime moulds. Isolated during this study were 14 species of dictyostelids. One species, Dictyostelium firmibasis is reported for the first time from Europe, and nine (Dictyostelium aureo-stipes, Dictyostelium sp. 1, Dictyostelium fasciculatum, Dictyostelium giganteum, Dictyostelium implicatum, Dictyostelium leptosomum, Dictyostelium sphaerocephalum, Polysphondylium candidum, Polysphondylium pallidum) are new records for the Mediterranean region. Members of the three genera Dictyostelium (ten species), Polysphondylium (three species) and Acytostelium (one species) are present in this region of the world. Comments on their diversity and distribution are included.     

Folic Acid as Second Chemotactic Substance in the Cellular Slime Moulds

IT has been known for some time that in certain species of cellular slime moulds acrasin, the substance which attracts the amoebae to central collection points during the aggregation phase, is cyclic AMP^1–4. We were also able to show that E. coli gave off another substance besides cyclic AMP (henceforth referred to as bacterial factor, or BF) which attracted the vegetative amoebae of Dictyostelium discoideum⁵. Here we demonstrate that this second attractant has the properties of folic acid or one of its derivatives. We also show that folic acid and related compounds not only attract the vegetative amoebae of D. discoideum (No. NC-4H) but also the amoebae of six other species (Dictyostelium rosarium No. CC-7; D. mucoroides No. 11; D. purpureum No. 2; D. minutum No. V-3; Polysphondylium violaceum No. 1; P. pallidum No. 2). For the latter three species cyclic AMP is not the aggregative attractant (ref. 6 and J. T. B., E. M. H., S. Noller, F. B. Oleson and A. B. Roberts, in preparation) which raises the interesting question of whether their acrasin might be related to the folates.     

Comparison of developmentally regulated lectins from three species of cellular slime mold

Extracts of the cohesive forms of the cellular slime molds Dictyostelium discoideum, Dictyostelium mucoroides and Dictyostelium purpureum contain lectin activity, assayed as hemagglutination activity. The lectin activity from each species binds quantitatively to Sepharose 4B and can be eluted with d-galactose. The resultant purified lectins are abundant proteins representing, in the case of D. purpureum, up to 5% of the total soluble protein of cohesive cells. The preparations from each species are similar but distinct in amino acid composition and other properties. Each purified preparation gives rise to two protein bands on sodium dodecyl sulfate-polyacrylamide gel electrophoresis with the major band representing as little at 77% (D. purpureum) and as much as 96% (D. mucoroides) of the total protein in the two bands. The molecular weights of the pair of bands were different for each species, ranging between about 23 000 and 26 000. The two bands are believed to represent subunits of lectins made up of either one or a combination of these two proteins. The apparent molecular weights of the purified lectin activities determined by sucrose density gradient centrifugation were all in the range of 100 000. $\text{N-}\text{Acetyl-}\text{d}\text{-galactosamine}$ was a potent inhibitor of the hemagglutination activity of each preparation; but there were some differences in the relative inhibitory potency of a number of other saccharides. Antiserum raised against each preparation, as well as univalent antibody fragments derived from these antisera, reacted best with the antigens to which they were raised; but showed some cross reaction measured both by precipitin reactions and by inhibition of hemagglutination activity of the purified lectins. The differences between the lectins from the different species could be trivial; but they also could be important for defining specific properties of these three species which reliably segregate into colonies of a single species when grown in mixed culture.     

Nuclear actin bundles in Amoeba, dictyostelium and human HeLa cells induced by dimethyl sulfoxide

In a previous study we demonstrated that dimethyl sulfoxide (DMSO) induces the formation of microfilament bundles in the interphase nucleus of a cellular slime mold, Dictyostelium mucoroides [12], in which the microfilaments bound rabbit skeletal muscle heavy meromyosin, forming an ‘arrowhead’ structure, and that this binding could be reversed by Mg²⁺ and ATP. In the present study, we show electron microscopic data demonstrating the occurrence of such microfilament bundles in the nucleus of Amoeba proteus and human HeLa cells, as well as in D. mucoroides. The similarities in the morphology and dimension of the microfilanets, as well as the specific conditions by which they are induced, suggested that these microfilaments are actin. We present evidence that actin is involved in interphase nucleus of a variety of organisms, and that DMSO acts on the molecules to induce microfilament bundles specifically in the nucleus.     

Reclassification of current Dictyostelium mucoroides strain Dm7 based on rRNA sequences and morphological features

Recent classification and phylogenetic analyses of dictyostelids are based on molecular sequences, but not all strains have been examined using this approach. Since the original identification in 1869, several strains called Dictyostelium mucoroides have been isolated; however, molecular taxonomic analyses of these strains remain insufficient. We therefore attempted to verify classification of one of these strains (Dm7) using rRNA sequences. As a result, strain Dm7 was found to be phylogenetically closer to D. clavatum and D. longosporum than to D. mucoroides. Comparisons of the morphological features of these strains further supported the classification based on sequence data, suggesting that Dm7 should be classified as a strain of D. clavatum.     

Phylogeography and sexual macrocyst formation in the social amoeba <Emphasis Type="Italic">Dictyostelium giganteum</Emphasis>

Background  

Microorganisms are ubiquitous, yet we are only beginning to understand their diversity and population structure. Social amoebae (Dictyostelia) are a diverse group of unicellular eukaryotic microbes that display a unique social behaviour upon starvation in which cells congregate and then some die to help others survive and disperse. The genetic relationships among co-occurring cells have a major influence on the evolution of social traits and recent population genetic analysis found extensive genetic variation and possible cryptic speciation in one dictyostelid species (Dictyostelium purpureum). To further characterize the interplay among genetic variation, species boundaries, social behaviour, and reproductive isolation in the Dictyostelia, we conducted phylogenetic analyses and mating experiments with the geographically widespread social amoeba Dictyostelium giganteum.     

Articular and vertebral lesions in the Pleistocene sloths (Xenarthra,Folivora) from the Brazilian Intertropical Region

We performed a macroscopic study on six species of Pleistocene sloths from the Brazilian Intertropical Region (BIR) including Valgipes bucklandi, Catonyx cuvieri, Mylodonopsis ibseni, Glossotherium sp., Ocnotherium giganteum and Eremotherium laurillardi and compared the results with a previous investigation of Nothrotherium maquinense and Ahytherium aureum. Differential diagnostic analysis revealed six varieties of musculoskeletal diseases: Calcium Pyrosphosphate Deposition Disease (CPPD), Spondyloarthropathy (SpA), Osteoarthritis (OA), Osteochondritis Dessicans (OD), Schmorl’s Node (SN) and Spondylosis Deformans (SD). We also carried out multivariate statistical analyses on all Pleistocene sloths to verify the similarity between taxa based on diagnosed diseases.     

Genetic Heterogeneity in Wild Isolates of Cellular Slime Mold Social Groups

This study addresses the issues of spatial distribution, dispersal, and genetic heterogeneity in social groups of the cellular slime molds (CSMs). The CSMs are soil amoebae with an unusual life cycle that consists of alternating solitary and social phases. Because the social phase involves division of labor with what appears to be an extreme form of “altruism”, the CSMs raise interesting evolutionary questions regarding the origin and maintenance of sociality. Knowledge of the genetic structure of social groups in the wild is necessary for answering these questions. We confirm that CSMs are widespread in undisturbed forest soil from South India. They are dispersed over long distances via the dung of a variety of large mammals. Consistent with this mode of dispersal, most social groups in the two species examined for detailed study, Dictyostelium giganteum and Dictyostelium purpureum, are multi-clonal.     

长白山苔原带凋落物生态化学计量特征及其对模拟氮沉降的响应

长白山苔原是我国乃至欧亚大陆东部独有的高山苔原,根据前人调查植被以灌木苔原为主要类型。在全球变暖背景下,近30年来,长白山岳桦林下的草本植物侵入苔原带,原生灌木苔原分化为灌木苔原、灌草苔原和草本苔原,形成了灌木、灌草混合和草本3种不同类型的凋落物,凋落物数量和质量发生显著改变。与此同时长白山苔原氮沉降量也在逐年增加,导致了土壤中氮的累积,势必影响凋落物的分解。凋落物作为连接植物和土壤的纽带,其分解过程中碳（C）、氮（N）、磷（P）等化学组分和化学计量比的变化直接和间接影响着土壤养分有效性和植物养分利用策略。为揭示氮沉降增加对长白山苔原带不同类型凋落物化学组分及生态化学计量特征早期变化的影响,开展了为期8个月的模拟氮沉降室内凋落物分解实验。在苔原带采集灌木优势种牛皮杜鹃和草本优势种小叶章的凋落物带回实验室,模拟灌木牛皮杜鹃群落、灌草混合的牛皮杜鹃-小叶章群落和草本小叶章群落的3种不同类型凋落物,设置三个施氮处理：对照（CK,0 g N m^-2 a^-1）、低氮（LN,10 g N m^-2 a^-1）、高氮（HN,20 g N m^-2 a^-1）。研究表明：（1）不施氮处理时,3种凋落物的C、P均呈释放状态,木质素（Li）呈先累积再略有降解趋势;牛皮杜鹃凋落物的N元素富集而其余两种凋落物N元素呈释放状态;灌草混合和草本凋落物比原生的灌木凋落物C和N元素释放快、Li累积少;而灌木凋落物的P释放略快于灌草和草本凋落物。3种植被类型凋落物的C/N、C/P、Li/N大小表现为：牛皮杜鹃凋落物>牛皮杜鹃-小叶章混生群落凋落物>小叶章凋落物;N/P表现为：小叶章凋落物>牛皮杜鹃凋落物>牛皮杜鹃-小叶章混生群落凋落物。（2）氮沉降促进3种类型凋落物分解过程中C、N和P化学组分的释放,且氮浓度越高促进作用越显著。在牛皮杜鹃凋落物分解过程中,氮素添加到达某一阈值后,其C/N、C/P、N/P、Li/N的降幅最大,后续若再增加氮素,其对化学计量比的影响均会减弱;本实验中的氮素添加量增加促进了小叶章凋落物的C/N、Li/N下降。（3）草本植物入侵引起凋落物类型的变化带来凋落物分解加快,将导致长白山苔原带养分循环的变化;氮沉降增加对小叶章凋落物化学组分的释放及C/N、Li/N的下降更为促进,小叶章凋落物内难分解化合物减少,分解受到促进。高氮沉降加快了小叶章凋落物与土壤、草本植物之间的养分循环。因此,随着未来苔原带氮沉降量的增加,将更有利于小叶章在与牛皮杜鹃的竞争中获胜,使苔原带呈现草甸化趋势。     

Rhododendron aureum Georgi formed a special soil microbial community and competed with above‐ground plants on the tundra of the Changbai Mountain,China

Rhododendron aureum Georgi is a perennial evergreen dwarf shrub that grows at all elevations within the alpine tundra of northern China. Previous research has investigated the plant communities of R. aureum; however, little information is available regarding interspecific competition and underground soil microbial community composition. The objective of our study was to determine whether the presence of R. aureum creates a unique soil microbiome and to investigate the relationship between R. aureum and other plant species. Our study site ranged from 1,800 to 2,600 m above sea level on the northern slope of the Changbai Mountain. The results show that the soil from sites with an R. aureum community had a higher abundance of nitrogen‐fixing bacteria and a higher resistance to pathogens than soils from sites without R. aureum. We emphasize that R. aureum promotes a unique soil microbial community structure that is distinct from those associated with other plants. Elevation and microbial biomass were the main influencing factors for plant community structure. Analysis of interspecific relationships reveals that R. aureum is negatively associated with most other dominant shrubs and herbs, suggesting interspecific competition. It is necessary to focus on other dominant species if protection and restoration of the R. aureum competition is to occur. In the future, more is needed to prove whether R. aureum decreases species diversity in the tundra ecosystems of Changbai Mountain.     

Interspecies and intraspecies interactions in social amoebae

The stable co‐existence of individuals of different genotypes and reproductive division of labour within heterogeneous groups are issues of fundamental interest from the viewpoint of evolution. Cellular slime moulds are convenient organisms in which to address both issues. Strains of a species co‐occur, as do different species; social groups are often genetically heterogeneous. Intra‐ and interspecies 1 : 1 mixes of wild isolates of Dictyostelium giganteum and D. purpureum form chimaeric aggregates, following which they segregate to varying extents. Intraspecies aggregates develop in concert and give rise to chimaeric fruiting bodies that usually contain more spores (reproductives) of one component than the other. Reproductive skew and variance in the proportion of reproductives are positively correlated. Interspecies aggregates exhibit almost complete sorting; most spores in a fruiting body come from a single species. Between strains, somatic compatibility correlates weakly with sexual compatibility. It is highest within clones, lower between strains of a species and lowest between strains of different species. Trade‐offs among fitness‐related traits (between compatible strains), sorting out (between incompatible strains) and avoidance (between species) appear to lie behind coexistence.     

Radial growth responses to gap creation in large,old Sequoiadendron giganteum

Questions: Do large, old Sequoiadondron giganteum trees respond to the creation of adjacent canopy gaps? Do other co‐occurring tree species and younger S. giganteum adjacent to gaps also respond? What are the likely factors affecting growth responses? Location: Mixed‐conifer forests of the southern Sierra Nevada, California, USA. Methods: We measured the growth response of large, old S. giganteum trees (mean DBH=164 cm; ages estimated >1000 yr) to gap creation by coring trees and comparing growth after gap creation to growth before gap creation. We also measured young Abies concolor, Pinus lambertiana, and young S. giganteum. Gap‐adjacent trees were compared with non‐adjacent reference trees. Tree rings were analysed for carbon isotope discrimination and for longer‐term growth trend correlations with climate. Results: Following gap creation gap‐adjacent old S. giganteum grew more than reference trees. Abies concolor trees also exhibited a growth response to gap creation. No response was detected for young S. giganteum or P. lambertiana, although detection power was lower for these groups. There was no difference in carbon isotope discrimination response to gap creation between gap‐adjacent and reference trees for old S. giganteum and radial growth was positively correlated with winter precipitation, but not growing season temperature. Conclusion: It is unclear what caused the growth release in old S. giganteum trees, although liberation of below‐ground resources following removal of competing vegetation appears to be a significant contributor. Sequoiadondron giganteum, the third‐longest lived and the largest of all species, remains sensitive to local environmental changes even after canopy emergence. Management activities that reduce vegetation surrounding individual specimen trees can be expected to result in increased vigor of even these very old and large trees.     

The ethylene action in the development of cellular slime molds: an analogy to higher plants

Summary The cellular slime moldDictyostelium mucoroides-7 (Dm 7) and its mutant (MF 1) exhibit sexual or asexual development depending upon culture conditions. During the sexual cycle macrocyst formation occurs, whereas sorocarps containing spores and stalk cells are asexually formed. As previously reported, the macrocyst formation is marked by the emergence of true zygotes, and is induced by a potent plant hormone, ethylene. The concentration of ethylene required for macrocyst induction was determined to establish the similarity of ethylene action between this organism and higher plants. Macrocysts are induced by low (1 l/l) exogenous concentrations of ethylene. Higher concentrations (10–1,000 ul/l) also gave essentially the same inductive activity. Ethionine, an analogue of methionine, was found to inhibit zygote formation during sexual development through its interference with ethylene production by Dm 7 and MF 1 cells. In fact, the inhibitory effect of ethionine was mostly nullified by the application of ethylene, S-adenosyl-L-methionine, or 1-aminocyclopropane-1-carboxylic acid. Taken together these results suggest that both the effective concentration of ethylene and the pathway of ethylene biosynthesis inD. mucoroides may be similar to those in higher plants. Ethylene was also found to be produced in various species and strains of cellular slime molds, even during the asexual process. The possible functions of ethylene in the asexual development are discussed in relation to cell aggregation and differentiation.Abbreviations SAM S-adenosyl-L-methionine - ACC 1-aminocyclopropane-1-carboxylic acid - AOA (aminooxy) acetic acid - BSS Bonner's salt solution - DAPI 4,6-diamidino-2-phenylindole