Comparative experimental tests of natricine antipredator displays, with special reference to the apparently unique displays in the Asian genus, Rhabdophis

Nuchal glands are unique organs known in only twelve Old World natricine species (three genera) including Rhabdophis tigrinus tigrinus. The glands of R. t. tigrinus contain a toxic secretion that may be derived from its toxic toad diet. A series of peculiar antipredator behaviors, for example neck arch, neck butt, and dorsal-facing posture, are exhibited by R. t. tigrinus, and a functional association between this behavior and the nuchal glands has been suggested. To investigate the ubiquitousness of these putatively unique displays among snakes, antipredator responses of 27 taxa of natricine snakes, both with and without nuchal glands, were studied using a common testing procedure. Three of four taxa with nuchal glands exhibited neck arch, neck butt, and dorsal-facing posture. None of the remaining 23 taxa, which do not possess the glands, showed neck arch and neck butt. Principal-components analysis indicated the association of the above three displays along with the behaviors termed neck flatten and head elevation, confirming the presence of a series of nuchal gland-related behaviors. These results support the assumption that the suite of peculiar displays is associated with the deterrent effects of the nuchal gland secretion.     

Is <Emphasis Type="Italic">Gammarus tigrinus</Emphasis> (Crustacea,Amphipoda) becoming cosmopolitan through shipping? Predicting its potential invasive range using ecological niche modeling

While the intensity of global shipping has increased dramatically over the last decades, species exchange between continents has likewise intensified. Ballast water of ships is recognized playing a major role in this process. Many of the larger sea ports have become bridgeheads for invasions. Ecological niche modeling is used to investigate the potential invasive range and high invasive risk ports of the North American amphipod Gammarus tigrinus. Sixty-two occurrences of G. tigrinus in its native range (North America) and 34 environmental data sets were compiled. Data on dispersal distances were used via ecological niche modeling to analyze the invasive potential of G. tigrinus. The invasive risk of large ports was analyzed according to modeling result, as well as their salinity in the main oceanic routes of the world. G. tigrinus had a rapid range extension on the British Isles and in the rest of Western Europe. Now it is invading the countries surrounding the Baltic Sea. Worldwide it has a vast potential invasive range. It has a high invasive risk for many large ports along the main oceanic routes, among which the ports of Shanghai, Buenos Aires and Montevideo have the highest invasive risk. G. tigrinus may become cosmopolitan through shipping, and this possibility is increasing. Particular emphasis should be placed on preventing human-mediated dispersal. Ports may be the first places G. tigrinus invades. This study can identify high invasive risk ports, especially those at risk of introduced North America species. More importantly, the water of large ports should be monitored regularly for exotic aquatic organisms that may survive temporarily or permanently.     

Characterization of protoplasts prepared from the edible fungus, Stropharia rugoso-annulata

Protoplast preparation and regeneration conditions of the edible fungus, Stropharia rugoso-annulata Farlow apud Murrill were studied, and the regenerated progenies were characterized in this study. The optimal condition for protoplast preparation was incubation of young mycelia with gentle shaking in 1.5%(w/v) Lywallzyme at 30 °C for 3 h. PGPM (potato/glucose/peptone/mannitol) was the most suitable regeneration medium. Served as osmotic stabilizer, sugars (mannitol and sucrose) were better than inorganic salts (MgSO₄) for clone development and growth. Pre-incubation of protoplasts in liquid regeneration medium resulted in a significantly decreased regeneration rate. Both dikaryotic isolates and monokaryotic isolates could be identified from protoplast-regenerated progenies, with a much higher frequency of monokaryotic isolates identified from the early-developed and fast-growing regenerated clones. Two parental mating types were also identified from protoplasted monokaryotic isolates, but not segregated by 1:1. The mycelial growth rate of protoplasted monokaryotic isolates showed a mating type-dependent model when cultured at different incubation temperatures and pH values, with A2B2 mating type monokaryotic isolates growing faster than those of A1B1 mating type monokaryotic isolates.     

Nuclear selection in oidium formation from dikaryotic mycelia ofFlammulina velutipes

The effect of nuclear dominance in monokaryotic oidium formation from dikaryotic mycelia in a tetrapolar basidiomycete,Flammulina velutipes, was examined. A total of 46 monokaryotic stocks were used to produce 194 hybrid dikaryotic stocks by crossing. The proportion of homokaryons among the oidium isolates from dikaryotic mycelia was over 95%. The staining of nuclei of oidia with propidium iodide showed that over 90% of oidia were monokaryotic and suggested that these oidia had single haploid nuclei at the G1 stage. The monokaryotic oidium isolates from hybrid dikaryons were backrossed to parental monokaryotic stocks. Although most of the monokaryotic oidium isolates (except for those from 17 hybrid dikaryons from a total of 194 test stocks) showed nuclear types similar to only one of the parental stocks, the process seems to produce essentially the split nuclear type composition. Therefore, the monokaryotization in oidium formation from dikaryotic mycelia essentially involves the process of nuclear selection. The two separate results of hierarchies of relative dominance among two nuclei of the parental dikaryons in the monokaryotic oidium formation by grouping with incompatibility factor compositions were determined. Only a few discrepancies were found in the hierarchies between the two specific nuclear compositions of hybrid dikaryons.     

Ectomycorrhiza formation between Pseudotsuga menziesii seedling roots and monokaryotic and dikaryotic isolates of Laccaria bicolor

Seedling roots of Pseudotsuga menziesii were colonized with three monokaryotic isolates and one dikaryotic isolate of Laccaria bicolor to assess the effect of fungal genotype on ectomycorrhiza formation. Ectomycorrhizas resulting from colonization by the dikaryotic isolate had a multilayered mantle and a cortical Hartig net. One monokaryotic isolate (ss7) formed ectomycorrhizas comparable in anatomy to those induced by the dikaryotic isolate. Two other monokaryotic isolates (ss5, ss1) failed to form mantles or Hartig nets. Roots colonized by these isolates developed characteristics indicating an incompatible reaction.     

Nuclear selection in monokaryotic oidium formation from dikaryotic mycelia in a basidiomycete,Pholiota nameko

The effect of nuclear dominance in monokaryotic oidium formation from dikaryotic mycelia inPholiota nameko was examined. Over 90% of oidium isolates from dikaryotic mycelia were monokaryotic. Although only one parental nuclear type was recovered from an average of about 80% in these isolates, the nuclear selection process in oidium formation seems essentially to produce split nuclear type composition in oidium products. The hierarchy of relative dominance among the nuclear types of the parental dikaryons in monokaryotic oidium formation was determined. The two hierarchies in nuclear selection between monokaryotic oidium formation and monokaryotic mycelium formation coincided at a level of at least 75%.     

Food selection in three leaf-shredding stream invertebrates

Leaf disks (Betula papyrifera) were conditioned for two weeks by six species of aquatic hyphomycetes. Mass losses of the leaves were determined, and their concentrations of protein (extracted at pH 7, 10 and 12.8), phenolics (Folin-Ciocalteu and BSA-precipitation), lipids, and ergosterol (as indicator of fungal biomass) were measured. Enzymatic activities of the culture filtrates against cellulose, xylan and pectin were estimated. Gammarus tigrinis, Pycnopsyche guttifer and Tipula caloptera were given a choice of the six leaf/fungus combinations. G. tigrinus and P. guttifer consistently preferred some combinations over others; T. caloptera appeared to feed randomly. There were no significant correlations between consumption and any of the measured characteristics of leaf disks. With G. tigrinus and P. guttifer, the sequence of preference could be reproduced by extracting mycelia with non-polar solvents and applying the extracts to unconditioned leaf disks. Consumption of extract-coated disks was lower than consumption of conditioned disks. Numbers of endosymbiotic gut bacteria increased from G. tigrinus to P. guttifer to T. caloptera; diet diversity showed the opposite trend.     

Germination and growth ofErythrorchis ochobinesis (Orchidaceae) accelerated by monokaryons and dikaryons ofLenzites betulinus andTrametes hirsuta

Four sib-monokaryons and two reconstituted dikaryons of two basidiomycetes,Lenzites betulinus andTrametes hiruta, accelerated the seed germination ofErythrorchis ochobiensis, an achlorophyllous orchid. All isolates ofL. betulinus and three isolates ofT. hirsuta induced the development of plants from germinated seeds. Although three monokaryotic isolates ofT. hirsuta failed to induce the development of plants, the reconstituted dikaryons induced the development.     

STUDIES ON THE MATING SYSTEMS OF ELEVEN SPECIES OF MYXOMYCETES

Genetic analysis of 20 isolates representing 11 species of Myxomycetes was carried out to determine whether they were heterothallic or homothallic. Each of the single isolates of Badhamia utricularis, Didymium minus, D. nigripes, and Physarum cinereum was found to be heterothallic, the first such reports for all of the species except D. nigripes. All isolates of Badhamia foliicola (1), Didymium squamulosum (1), Physarum compressum (3), P. gyrosum (1), P. pusillum (2), Stemonitis flavogenita (6) and an unidentified species of the order Physarales (2) were homothallic, with B. foliicola, P. compressum, and S. flavogenita being reported for the first time.     

ITS rDNA variation of the Coprinopsis phlyctidospora (syn.: Coprinus phlyctidosporus) complex in the Northern and Southern Hemispheres

 Coprinopsis phlyctidospora (syn: Coprinus phlyctidosporus) from the Netherlands, Japan, New Zealand, and Australia can be segregated into two groups, northern and southern, based on the nucleotide sequences of their ITS regions. The mating type of a C. phlyctidospora isolate was tetrapolar. Mating reactions were compatible between monokaryotic testers derived from basidiospores of a Japanese isolate and dikaryotic isolates obtained from a wide geographic area in Japan. In contrast, mating between the Japanese monokaryotic and dikaryotic isolates from Australia and New Zealand were incompatible. These results indicated that C. phlyctidospora was complex and individuals currently recognized as C. phlyctidospora in the Northern Hemisphere and those in the Southern Hemisphere are distinct taxa. The relationship between the clades and the biogeography of the C. phlyctidospora complex are also discussed. Received: January 22, 2002 / Accepted: March 15, 2002     

Seasonal variability in the grazing potential of the invasive amphipod Gammarus tigrinus and the native amphipod Gammarus salinus (Amphipoda: Crustacea) in the northern Baltic Sea

Mesograzers are known to reduce the biomass of their host plant and modify the structure of the whole macrophyte community in many ecosystems. Thus, the introduction of an efficient mesograzer may destabilize macrophyte community and also affect the native grazers. We estimated how large proportion of macrophyte production are consumed by the alien gammarid G. tigrinus and the native gammarid G. salinus in the species poor ecosystem of the northern Baltic Sea. We analysed whether G. tigrinus consumes different diet as the native G. salinus and whether the effect of G. tigrinus on the survival of the native G. salinus is macrophyte species specific. Grazing experiments showed that there was a clear difference in the grazing rates of gammarids among the studied macrophyte species in summer and autumn but not in spring. The grazing rates were significantly higher in the prevailing macrophyte Pilayella littoralis as compared to other macrophytes. The grazing was inversely related to the diurnal net photosynthetic values of macrophytes. The gammarid amphipods potentially removed only a minor part of plant primary production except for summer and autumn when grazing of a few perennial species exceeded macrophyte production. Macrophyte species and presence of G. salinus had no effect on the survival of G. tigrinus. The presence of G. tigrinus, however, reduced the survival of the native gammarids within P. littoralis in summer. To conclude it is likely that both native and alien gammarid amphipods do not exert significant pressure on the macroalgal communities in the northern Baltic Sea. Competitive interactions between G. tigrinus and G. salinus within the prevailing macrophyte P. littoralis is the likely explanation of the decline of the native gammarid amphipods after the establishment of G. tigrinus in the northern Baltic Sea.     

Genetic analysis of nuclear ribosomal DNA of Lentinula edodes

Genetic analysis of nuclear ribosomal DNA (rDNA) of Lentinula edodes was carried out using rDNA restriction fragment length polymorphisms (RFLPs) as genetic markers. Two compatible monokaryotic strains that differed in the endonuclease digestion patterns of their rDNA were used. The dikaryotic strain established by crossing them produced mixed RFLP patterns. Single-spore isolates derived from the dikaryotic strain showed three types of rDNA RFLP patterns: either one of the two parental types or a mixed type. From the frequency of the mixed type, the recombination value of rDNA tandem repeats was calculated to be 31.4%. Linkage analysis between rDNA and two incompatibility factors (A and B) revealed that rDNA was not linked to either factor. The rDNA genotypes did not affect mycelial growth among the single-spore isolates.     

The role of laccase and peroxidase of <Emphasis Type="Italic">Lentinus (Panus) tigrinus</Emphasis> fungus in biodegradation of high phenol concentrations in liquid medium

The possibility of the usage of Lentinus tigrinus fungus strain VKM F-3616D for biodegradation of high (up to 5%) phenol concentrations in liquid medium and the involvement of laccase and peroxidase in this process have been studied. L. tigrinus fungus was demonstrated to effectively degrade phenol with easy biomass deletion from the liquid. Decrease in phenol concentration was accompanied by increased secretion level and laccase activity at the preliminary stages of biodegradation, while that of peroxidase was at the latest stages of biodegradation. These enzyme secretions in distinct ratios and consequences are necessary for effective phenol biodegradation. An effective approach for phenol concentration decrease in the waste water of smoking shops in meat-processing factories using L. tigrinus fungus was described.     

Mating system and DNA polymorphism of monokaryons with different mating type of Stropharia rugoso-annulata

The mating system of Stropharia rugoso-annulata Farlow apud Murrill was studied by pairing single spore isolates from the same fruitbody, and the genetic diversity of monokaryotic strains with different mating types was evaluated by the RAPD technique. Basidiospores could germinate normally on PGP (potato/glucose/peptone) medium at 30 °C. Analysis of self- and cross-pairings revealed that Stropharia rugoso-annulata was heterothallic and tetrapolar. RAPD analysis detected polymorphism among monokaryotic strains, with more genetic variation within monokaryotic strains with non-parental mating type compared to monokaryotic strains with parental mating type. These results were in general agreement with the existing knowledge, confirming the validity and usefulness of the RAPD technique. Therefore, the RAPD technique will provide an exciting and valuable tool for a large-scale study on identification and genetic resources of monokaryotic strains, and should lead to a more efficient understanding and utilization of genetic diversity of monokaryotic strains in cross breeding by breeders.     

Geographic distribution and food habits of Leopardus tigrinus and L. geoffroyi (Carnivora,Felidae) at their geographic contact zone in southern Brazil

The objective of this study is to define the geographic distribution of Leopardus tigrinus and L. geoffroyi in one of the few regions of South America where they co-occur, the state of Rio Grande do Sul (RS), in southernmost Brazil. We compiled 133 records for both species and constructed a distribution map, which shows sharp geographic segregation between them. Leopardus tigrinus was found to be associated more with forested ecoregions in the northern part of the state, while L. geoffroyi records were mainly associated with open habitats of the Pampas biome in southern RS. We present data on the diet of these two species that indicate trophic niche separation between them in this region of geographic contact. Our results thus suggest that these species exhibit ecological partitioning with respect to habitat and prey, and that these factors may influence the observed pattern of limited spatial overlap in this region.     

Interference competition between alien invasive gammaridean species

The relative abundances of gammaridean species in the river Rhine have profoundly changed since the invasion of Dikerogammarus villosus in 1994/1995. This study tested whether these changes in gammaridean dominance could have been determined by interspecific competition and unequal mortality, for example by intraguild predation (IGP). Single and two species tests have been carried out in aquariums provided with all substrata present in the main channel of the Rhine. Changes in substratum choice, increased swimming activity and increased mortality of a species were used as indicators of interspecific competition during interaction between gammaridean species. Interspecific competition and mortality between the most abundant invasive gammaridean species in the Rhine, viz. Gammarus tigrinus, Echinogammarus ischnus and Dikerogammarus villosus were tested. In single-species experiments, G. tigrinus and D. villosus showed similar preferences for a stony substratum, whereas E. ischnus mostly occupied the water column. The two-species aquarium experiments indicated direct interference competition for substratum and unequal mortality between G. tigrinus and D. villosus, with D. villosus being the stronger competitor. Competitive stress was influenced by population density, was size-dependent and varied between the different types of substratum due to substratum choice. G. tigrinus did not show any behaviour indicative of interference competition in the presence of E. ischnus, and neither did E. ischnus or D. villosus in the presence of any of the other gammarideans. Swimming in the water layer may already enable E. ischnus to minimise its encounters with the stone-dwelling D. villosus and G. tigrinus. To maximise the encounters between E. ischnus and D. villosus, a fish (Lepomis gibbosus) was added to occupy the water layer during the aquarium experiments. E. ischnus showed a higher mortality in the presence of both D. villosus and fish, probably due to increased stress, as shelter opportunities to escape the predators had been minimised. The study shows that interference competition between gammaridean species can explain the replacement of the North American invader G. tigrinus by D. villosus in the river Rhine. E. ischnus and D. villosus both Ponto-Caspian invaders did not show interference competition in our experiments and co-exist in the Rhine.     

The secotioid form of Lentinus tigrinus: genetics and development of a fungal morphological innovation

Secotioid fungi resemble gasteromycetes, but are presumably closely related to agaricoid fungi. Lentinus tigrinus is a wood-decaying mushroom that has both a secotioid and an agaricoid form. We examined ontogeny and heritability of the secotioid phenotype in L. tigrinus with a combination of formal genetic crosses, scanning electron microscopy, and macroscopic observation of cultured sporocarps. For F1 analysis, we crossed single-spore isolates (SSIs) representing four mating types derived from a secotioid dikaryon and four mating types from an agaricoid dikaryon. All F1 sporocarps had typical agaricoid morphology. For F2 analysis, 200 SSIs from one F1 sporocarp and 100 SSIs from another F1 sporocarp were backcrossed to tester SSIs from sporocarps produced by the parental secotioid dikaryon. Ratios of secotioid to agaricoid dikaryons thus produced were 47:49 and 84:109, which confirms previous reports that the secotioid phenotype is conferred by a recessive allele at a single locus (χ² = 0.0417, P > 0.05, χ² = 3.2383, P > 0.05, respectively). Early ontogeny of the secotioid form is indistinguishable from that of the agaricoid form. Later, the hymenophore is obscured by a weft of hyphae that proliferates from the margins of the developing lamellae. Longevity of the sporocarps and rate and duration of sporocarp growth are approximately equal in the secotioid and agaricoid forms. Developmental evolution of the secotioid form is interpreted as an example of von Baerian differentiation, rather than paedomorphosis, which has been implicated in evolution of other secotioid taxa.     

Population dynamics of alien gammarid species in the River Odra estuary

During the period of 1988–2000, three alien gammarid species, i.e. Gammarus tigrinus Sexton, 1939, Pontogammarus robustoides G. O. Sars, 1894, and Dikerogammarus haemobaphes (Eichwald, 1841) were recorded in the Odra estuary. This study, the first record of G. tigrinus and P. robustoides in Poland, focuses on the distribution, abundance, and biomass as well as on population structure of the three new species found in the coastal zone of the estuary. G. tigrinus and P. robustoides were first sampled in the Szczecin Lagoon (mid-part of the estuary) in 1988. The densities of both species were high from the time they were first recorded and exceeded 10 000 ind. m^–2, while the biomass was 25 and 140 g m^–2, respectively. G.tigrinus occurred at higher densities in northern and eastern parts of the Lagoon, while P. robustoides was particularly abundant in the south-eastern part, most heavily affected by River Odra discharge. In 1999, the third gammarid, D. haemobaphes, was recorded in the southern part of the estuary. Both in 1999 and 2000, the species was found in qualitative samples (dredge collections of Dreissena polymorpha) taken from the western branch of the river. As estimated from quantitative samples, densities and biomass in the eastern branch were rather low and did not exceed 150 ind. m^–2 and 2.7 g m^–2, respectively. The Szczecin Lagoon population of G. tigrinus produces at least two generations within a year: a summer one and an overwintering one. The latter is dominated by older and larger specimens. The species may reproduce in the Lagoon from April until November.     

Pigmentation and sporulation in selected Myxomycetes

Chemical, chromatographic and spectrometric methods are used to characterize plasmodial pigments and determine relationships between pigmentation and sporulation in selected Myxomycetes. In Physarum gyrosum (white) a single pigment is identified and characterized as a flavone. Physarum polycephalum (yellow) and Didymium iridis (brown) contain four and six components, respectively, in their plasmodial pigments which test negatively for flavones but show the presence of some type of phenolic compound. No detectable component is identified in the white plasmodium of Didymium squamulosum which proved to be independent of light for fruiting. The absorption spectra of all species that were light sensitive for fruiting showed common peaks in the 300–400-mμ region, among others. Pigment changes associated with light absorption are reported for some white, yellow and brown plasmodial types. In Physarum gyrosum a yellow pigment forms in light which did not show the characteristic flavones present in the white plasmodial stage. Changes in absorption spectra are reported for Physarum polycephalum, Didymium iridis and Didymium squamulosum as the plasmodial pigments change prior to fruiting. Results show a close relationship between the physiology of plasmodial pigmentation and sporulation in the Myxomycete species studied.     

Effect of Ethanol on Enzymatic Activity of Fungal Laccases

Blue laccase from Coriolus versicolor and blue and yellow laccases from Panus tigrinus were isolated, purified and studied in acetate buffer solutions, with and without addition of various amounts of ethanol, using syringaldazine and 2,6-dimethoxyphenol as substrates. Effect of ethanol on blue laccases could be successfully described using the mixed inhibition model, over the range of 0–2.5 M ethanol concentrations. Yellow laccase from P. tigrinus behaves differently, which may be explained by the presence of some extra molecules in its structure, which possibly stabilize the enzyme and might be exchanged in ethanol solutions.