Kalkbohrende Mikrothalli beiHelminthocladia undScinaia (Nemaliales,Rhodophyta)

Shell-boring microthalli in Helminthocladia and Scinaia (Nemaliales, Rhodophyta). Spores shed from pink mussel shells were shown to develop into branched monosiphonous thalli, their filaments penetrating into shell fragments. Isolates from four single germlings were cultivated. Two of these produced gametophytes ofHelminthocladia andScinaia; the others have so far only reproduced by tetraspores or monospores. Evidently the microthalli of some genera of the Nemaliales — which are, with the exception ofNemalion multifidum, known only from cultures — are shell-inhabiting and have therefore not been found in nature. The adult algae occur mainly on shells and CaCO₃ substrates. Until the beginning of the century,Helminthocladia andScinaia frequently occurred at Helgoland, but they have not been found there for more than 50 years. Their microthalli, however, are still present as shell-boring algae. This study is intended to stimulate similar ones in other genera of the Nemaliales so as to obtain a broader basis for discussion of systematic and phylogenetic relationships.     

RELATION OF TEMPERATURE TO DEVELOPMENT OF THE MACROTHALLUS OF DESMOTRICHUM UNDULATUM1,2

Desmotrichum undulatum (J. Agardh) Reinke (Phaeophyta) was found in the winter months in the littoral zone of a tidal marsh in Virginia. When field collections were maintained in culture media at 21 C, most cells of the thallus developed into plurilocular sporangia. Released zoospores developed into microthalli from which clonal cultures were established. At 21 C microthalli produced sporangia which released zoospores, subsequently developing into more microthalli. However, microthalli at 6 C formed filaments which subsequently developed into thalloid plants. When the thalloid plants of Desmotrichum formed in culture were subjected to 21 C, plurilocular sporangia were formed. These results support the proposition that the seasonal periodicity of Desmotrichum is due to a temperature-regulated phenomenon in the development of macrothalli.     

CULTURE STUDIES ON REPRODUCTION OF SPERMATOCHNUS PARADOXUS (PHAEOPHYCEAE,CHORDARIALES)1

The life history of Spermatochnus paradoxus (Roth) Kütz. isolated from the Mediterranean Sea was studied in culture. Meiospores develop to a microscopic stage (microthallus) which at 20°C perpetuates asexually by plurilocular sporangia and formation of new microthalli. At 9°C microthalli act as homothallic gametophytes. Fusion of isogametes results in a diploid microthallus which, after differentiation of an apical cell, leads back to Spermatochnus plants. In addition, gametes develop without fusion to form haploid macrothalli, the further fate of which has not been determined. Chromosome numbers alternate between n = 20 ± 2 in the microthalli and 2n = 41 ± 4 in macrothalli.     

A Comparative Study of the Infraciliature and Silverline System of the Fresh-Water Scuticociliates Pseudocohnilembus putrinus (Kahl, 1928) nov. comb., P. pusillus (Quennerstedt, 1869) nov. comb., and the Marine Form P. marinus Thompson, 19661

Three species of Pseudocohnilembus were examined with respect to infraciliature and the silverline system. The freshwater species P. putrinus and P. pusillus exhibited no differences in general organization of the argyrophilic structures when compared with the marine P. marinus. A review of the Pseudocohnilembus species that have been described on the basis of silver preparations shows that at present four species have been well characterized: P. pusillus (Quennerstedt, 1869) nov. comb., P. putrinus (Kahl, 1928) nov. comb., P. hargisi Evans & Thompson, 1964, and P. marinus Thompson, 1966, P. persalinus Evans & Thompson, 1964 and P. longiseta Evans & Thompson, 1964 are regarded as identical with the older known P. pusillus, because with respect to morphology and argyrophilic structures they are within the range of variability of that species; and their names thus fall as synonyms of P. pusillus.     

Using DNA‐based techniques to identify hybrids among linear‐leaved Potamogeton plants collected in China

Abstract It is well known that interspecific hybrids occur in the genus Potamogeton. The linear‐leaved Potamogeton species commonly have highly variable morphological characteristics. Their hybrids often show similar vegetative characters to their parental species and their identification based solely on morphology is not always conclusive. In order to clarify whether there are any hybrids from the linear‐leaved Potamogeton plants collected in China, we used internal transcribed spacers (ITS) of nuclear ribosomal DNA and chloroplast rbcL gene sequences to identify the hybrids. Using ITS sequence additivity, we identified four hybrids, namely P. orientalis (P. pusillus×P. oxyphyllus), P. pusillus×P. berchtoldii, P. foliosus×P. octandrus, and P. cristatus×P. octandrus. The latter three hybrids should be considered as new hybrids in Potamogeton. The maternal parents of the four hybrids were confirmed using chloroplast rbcL gene sequences.     

Influence of culture conditions on the production of milk-clotting enzyme from Rhizomucor

The ratio of milk-clotting activity to proteolytic activity (MC/PA) was used as an index to determine the quality of milk-clotting enzyme. Solid-state fermentation on wheat bran for 5 days at room temperature gave optimal production for enzyme by Rhizomucor miehei and R. pusillus. A ratio of wheat bran to moisture of 1:0.6 (w/v) gave best results. Adding skim milk powder to the media of R. miehei did not improve the MC/PA ratio but 4% (w/w) of the powder did improve the ratio with R. pusillus. Co-cultivation of R. miehei with R. pusillus did not change the MC/PA ratio.The authors are with the Department of Biotechnogy, Bharathiar University. Coimbatore-641 046 India     

Induced azygospore formation in Mucor (Rhizomucor) pusillus by Absidia corymbifera

Attempts to determine the mating reaction type of heterothallic strains of Mucor pusillus in interspecific contrasts with Mucor strains of known mating reaction type were unsuccessful. Contrasts with Absidia corymbifera strains resulted in the production of azygospores in Mucor pusillus.     

Integrative transformation of the zygomycete Rhizomucor pusillus by homologous recombination

 For development of a homologous transformation system for the zygomycete fungus, Rhizomucor pusillus, the isopropylmalate isomerase (leuA) gene was cloned from R. pusillus IFO 4578 by the DNA-probing method with the leuA sequence of Mucor circinelloides as probe. The nucleotide sequence revealed that leuA of R. pusillus encoded a 755-amino-acid protein of 82.5 kDa with no intron. The leuA gene on pUC19 (plasmid pRPLeu10) was introduced by polyethyleneglycol-assisted transformation into protoplasts of a leuA ^- mutant of R. pusillus that was obtained by UV mutagenesis. Transformation under optimal conditions yielded 20 Leu⁺ transformants (μg pRPLeu10 DNA)^-1 (1×10⁶ viable protoplasts)^-1. Blot analysis of DNA from the transformants showed that the pRPLeu10 sequence was integrated into the genome by homologous recombination at the leuA locus. Received: 2 October 1995/Received last revision: 5 December 1995/Accepted: 11 December 1995     

Genetic variation within and between populations of Potamogeton pusillus agg.

Patterns of isozyme variation were examined in 17 populations of P. pusillus and P. berchtoldii, together with one population of P. trichoides taken for comparison. Both P. pusillus and P. berchtoldii displayed low levels of variation within populations associated with high levels of interpopulation differentiation. This pattern of partitioning of genetic variation within and between populations is attributed to the founder effect, frequent vegetative propagation by turions, dominant self-fertilization and limited seedling recruitment. The mechanism of pollen transfer was investigated in cultivation. Effective pollination takes place in air above the water surface (autogamy, geitonogamy, anemogamy), on the water surface (epihydrogamy) or below water surface (hydroautogamy). The species are self-compatible. The low level of infra-population variation together with rare occurrence of heterozygotes suggest that selfing is the most frequent mode of pollination, although the protogynous flowers may occasionally permit some cross-pollination. Unique enzyme markers were found for P. pusillus and P. berchtoldii, and also for the single population of P. trichoides. All multienzyme phenotypes were species-specific. Isozyme data support the separate position of P. pusillus and P. berchtoldii. UPGMA dendogram based on enzyme data of 133 plant samples revealed three distinct main enzymatic entities perfectly corresponding to the three morphologically defined species.     

Genetic transformation of a Rhizomucor pusillus mutant defective in asparagine-linked glycosylation: production of a milk-clotting enzyme in a less-glycosylated form

Rhizomucor pusillus 1116R3 has a defect in alg2 encoding a mannosyltransferase in the asparagine (N)-linked oligosaccharide biosynthetic pathway and produces proteins in less-glycosylated forms. For development of a genetic transformation system for this zygomycete, an uracil auxotroph (mutant 1116U17) as the host strain was derived by ultraviolet (UV) mutagenesis as 5-fluoroorotic acid-resistant colonies and the orotidine-5′-monophosphate (OMP) decarboxylase (pyr4) gene as a selection marker was cloned from the wild-type strain R. pusillus F27 by the polymerase chain reaction with primers designed on the basis of the pyr4 sequences from other fungi. The amino acid sequence of R. pusillus Pyr4 deduced from the nucleotide sequence showed high homology with the OMP decarboxylases from various fungi. The pyr4 gene on pUC19 (plasmid pRPPyr4) was introduced into protoplasts of R. pusillus 1116U17 by polyethylene glycol-assisted transformation. Transformation under optimized conditions yielded 5 Ura⁺ transformants with 1 μg pRPPyr4 DNA and 1 × 10⁷ viable protoplasts. Southern blot analysis of the genomic DNA from the transformants showed that multiple copies of the pRPPyr4 sequence were integrated into the genome by homologous recombination at the pyr4 locus. For the purpose of production of a milk-clotting aspartic proteinase (MPP) in a less-glycosylated form, mpp from the wild-type strain was cloned in pRPPyr4 and introduced into protoplasts of R. pusillus 1116U17. Transformants obtained in this way contained multiple copies of mpp at the chromosomal mpp locus and produced MPP as a mixture of molecules having no sugar chains and Man_0∼1GlcNAc₂ at the two N-linked glycosylation sites in an amount about 12 times larger than the parent strain. The transformation system for R. pusillus 1116U17 would be useful for production of proteins with truncated N-linked oligosaccharide chains. Received: 1 February 1999 / Received revision: 26 February 1999 / Accepted: 20 March 1999     

Performance of the predator Delphastus pusillus on Bemisia resistant and susceptible tomato lines

Host plant resistance and biological control are often assumed to act additively to suppress populations of agricultural pests. Using tomato trichome based resistance to the whitefly Bemisia argentifolii, we tested this additivity assumption with Delphastus pusillus, a coccinelid predator of Bemisia. Various life history traits of D. pusillus were measured on the tomato cultivar Alta, which possessed foliage with 3-fold greater trichome densities than the second cultivar VF145B7879. Beetles housed on VF145B7879 exhibited significantly greater lifetime fecundities and walking speeds than beetles housed on Alta. No cultivar-specific differences were observed in D. pusillus longevities or handling times. Combining these observations with previously published reports of reduced B. argentifolii population growth rates on Alta compared to VF145B7879, we tested the hypothesis that increased levels of whitefly biological control could be obtained on VF145B7879 by comparison to Alta through releases of D. pusillus. Analyses of results obtained from replicated population trials detected significant reductions in whitefly populations due to D. pusillus releases, but they did not detect a significant influence of tomato cultivar on the ability of D. pusillus to suppress whitefly populations. Significantly longer beetle residence times on Alta than on VF145B7879 may have compensated for the significantly slower walking speeds and reduced lifetime fecundities observed on Alta and produced a neutral effect of foliar trichome densities on B. argentifolii biological control.     

Über die Variabilität von Stephanodiscus hantzschii GRUN

A new variety, striatior, of the diatom Stephanodiscus hantzschii Grun. (S.h.) with denser striae was described (Kalbe 1971). The legality of S.h. var. pusillus is confirmed by specimens from freshwaters of the north of Mecklenburg. The species S.h. probably consists in this region of several races with different peaks of valve diameter variation. The planctonic mass changing of S.h. and its forms in the three rivers Warnow, Malchiner Peene and Neukalener Peene and in the Lake Kummerow is represented. The differences of the average valve sizes in these waters are remarkable. Var. pusillus Grun. appears to be a benthic mass form, too. High cell numbers and high cell volume sums are not developing ever simultaneously. Var. pusillus is not a suitable form for evaluating the saprobiological status, for it is adaptable to different ecological conditions. The species S.h. itself is a beta- to alphamesosaprobic organism, being able to produce high cell numbers within this range of saprobity.      

Regeneration and colonization abilities of aquatic plant fragments: effect of disturbance seasonality

The regeneration (regrowth into viable plants or production of propagules, such as turions or buds) and colonization (development of roots and establishment in the sediment) of fragments of six aquatic plant species (Elodea canadensis Michaux, Hippuris vulgaris L., Luronium natans (L.) Rafin., Potamogeton pusillus L., Ranunculus trichophyllus Chaix, Sparganium emersumRehmann) occurring in habitats frequently disturbed by floods, were investigated through laboratory experiments conducted in two seasons, and compared to the recolonization patterns depicted after field experiments. Hypothesis was that differences observed between recolonization patterns after spring and autumn flood disturbances should be related to differences in recolonization (via rooting) and regeneration (via propagules) abilities of species fragments. In May and in August, five types of fragments were collected from the plants. Their development and/or rooting abilities were recorded over 10 weeks in the greenhouse. Fragments from E. canadensisand H. vulgarishad higher regeneration and lower colonization abilities in spring and conversely in autumn. Fragments from R. trichophyllusand S. emersumhad high colonization and low regeneration abilities during both seasons. Fragments from L. natans developed new buds in spring, whereas root development occurred only in autumn. Fragments from P. pusillus never rooted into the sediment, but developed turions in autumn. Differences between recolonization patterns observed in the field at the two seasons can most often be related to differences in regeneration and colonization abilities of species fragments. Species that colonize disturbed areas rapidly whenever the disturbance by flooding occurs have at least one type of vegetative fragment with a high colonization potential; this is called the `always-ready strategy' which appears to be an adaptation of aquatic plants to the unpredictability of flood disturbances.     

Phytofiltration of As3+, As5+, and Hg by the aquatic macrophyte Potamogeton pusillus L,and its potential use in the treatment of wastewater

The aim of this paper was to investigate the capacity of the aquatic macrophyte Potamogeton pusillus to remove As³⁺, As⁵⁺, and Hg from aqueous solutions. The plants were exposed to 0 mg.L^?1, 0.1 mg.L^?1, 0.5 mg.L^?1, 1 mg.L^?1, or 2 mg.L^?1 of As³⁺, As⁵⁺, and Hg for 20 days. The results obtained for the individual removal of As³⁺, As⁵⁺, and Hg from water solutions, together with their accumulation in P. pusillus, indicate that this plant can be effectively used for the removal of Hg and of moderate concentrations of As³⁺ or As⁵⁺ (0.1 mg.L^?1) from aquatic systems. Roots and leaves accumulated the highest amount of As when the plant was exposed to As⁵⁺, but when it was exposed to As³⁺, the root accumulated the highest amount of As, and the leaves, the highest amount of Hg. When compared to other aquatic plants species, the results showed that P. pusillus demonstrated a higher Hg accumulation (2465 ± 293 µg.g^?1) when the transfer coefficient was 40,580 ± 3762 L.kg ^?1, showing the great potential of this macrophyte for phytoremediation of water contaminated with Hg. To the extent of our knowledge, this is the first report on bioaccumulation of As³⁺, As⁵⁺, and Hg by P. pusillus.     

Taxonomic review of Scinax fuscomarginatus (Lutz, 1925) and related species (Anura; Hylidae)

Scinax fuscomarginatus, Scinax parkeri, Scinax trilineatus, Scinax lutzorum, and Scinax pusillus are morphologically similar species with controversial taxonomy associated with open formations of South America east of the Andes. We used external morphology, advertisement call, and molecular sequences to assess the taxonomy of these species. Phylogenetic analysis of molecular data showed a well‐supported monophyletic group divided into two main clades and several subclades. Specimens of Hyla madeirae (synonym of S. fuscomarginatus) and those from Serra do Cachimbo correspond to distinct subclades, whereas the currently valid species were grouped into a single clade. This clade showed a marked substructure, but concordance of the recognized species to subclades was limited, showing S. fuscomarginatus to be paraphyletic with respect to S. parkeri, S. trilineatus, S. lutzorum, and S. pusillus. The high intrapopulation and intraspecific variation found in morphological characters did not allow us to differentiate amongst the currently recognized species. Similarly, none of them was distinguishable by morphometric analysis or advertisement call. Based on multiple evidence, we propose the synonymy of S. parkeri, S. trilineatus, S. lutzorum, and S. pusillus with S. fuscomarginatus, the revalidation of H. madeirae, and describe the specimens from Serra do Cachimbo as a new species. © 2014 The Linnean Society of London     

Reproductive ecology of three ocypodid crabs I. The influence of activity differences on reproductive traits

Reproductive traits of three ocypodid crabs,Scopimera globosa, Ilyoplax pusillus andMacrophthalmus japonicus, were compared.S. globosa andI. pusillus, inhabiting the upper-middle intertidal zone, produced 1–2 large broods per year, whereasM. japonicus, inhabiting the lower intertidal zone, produced 4–5 small broods per year. InS. globosa andI. pusillus, ovigerous females remained in their plugged burrows without feeding until their eggs hatched. On the other hand, femaleM. japonicus fed actively on surface mud while incubating. We concluded that few large broods may be advantageous in crab species that incubate in burrows, whereas continuous small broods may be advantageous in species that feed actively while incubating.     

2C DNA variation and relationships among New World species of the genus Lupinus (Fabaceae)

The 2C DNA values in 38 species and accessions of the genus Lupinus (Fabaceae) from the New World have been analysed using flow cytometry. They are representatives of North and South American species (the Atlantic and the Andean regions). Estimated 2C DNA values ranged from 1.08 pg in L. pusillus to 2.68 pg in L. albicaulis (both from North America), that is a variation of more than 2.5-fold. The variation for North American lupins was much higher than that for South American ones. Statistical analysis of the data resulted in a grouping that showed for North American lupins some correlation with the length of life cycle. Discussion concerns some aspects of the evolution of the genus.     

Changes in Aquatic Macrophytes after Liming Thrush Lake, Minnesota

Thrush Lake, Minnesota, was treated with limestone in 1988 to evaluate the efficacy of protective base addition against the loss of sport fisheries in a sensitive, mildly acidic lake. Prior to treatment, the lake was stressed (pH 6.46, ANC 64 μeq/L) but not severely degraded by acidic deposition and had a macrophyte community typical of lakes in northeastern Minnesota with low acid-neutralizing capacity (ANC). This paper describes the changes observed in aquatic plant communities during the 5 years after treatment, as pH and ANC slowly returned to pretreatment levels. Sphagnum platyphyllum, intolerant of non-acid conditions, was completely eliminated from the lake. The charo-phyte, Nitella, that originally shared dominance in the deep littoral zone with S. platyphyllum, decreased in importance during the first 2 years after treatment. Two vascular plants, Potamogeton pusillus and Najas flexilis, were first found in the lake the year after treatment and were abundant for 2 years after liming, probably in response to a combination of more neutral pH and reduced cover of Nitella. As the ANC and pH slowly returned to pretreatment conditions, Nitella again increased in coverage and depth range, with a concomitant decrease in P. pusillus and N. flexilis. The moss, S. platyphyllum, had not reinvaded the lake by 1993, 2 years after its dramatic decline.     

Molecular analysis of NAD+-dependent xylitol dehydrogenase from the zygomycetous fungus Rhizomucor pusillus and reversal of the coenzyme preference

The zygomycetous fungus Rhizomucor pusillus NBRC 4578 is able to ferment not only d-glucose but also d-xylose into ethanol. Xylitol dehydrogenase from R. pusillus NBRC 4578 (RpXDH), which catalyzes the second step of d-xylose metabolism, was purified, and its enzymatic properties were characterized. The purified RpXDH preferred NAD⁺ as its coenzyme and showed substrate specificity for xylitol, d-sorbitol, and ribitol. cDNA cloning of xyl2 gene encoding RpXDH revealed that the gene included a coding sequence of 1,092?bp with a molecular mass of 39,185?kDa. Expression of the xyl2 in R. pusillus NBRC 4578 was induced by d-xylose, and the expression levels were increased with accumulation of xylitol. The xyl2 gene was expressed in Escherichia coli, and coenzyme preference of the recombinant RpXDH was reversed from NAD⁺ to NADP⁺ in the double mutant D205A/I206R by site-directed mutagenesis.