Band 62, Heft 6

It was shown that compensatory base changes (CBCs) in internal transcribed spacer 2 (ITS2) sequence-structure alignments can be used for distinguishing species. Using the ITS2 Database in combination with 4SALE — a tool for synchronous RNA sequence and secondary structure alignment and editing — in this study we present an in-depth CBC analysis for placozoan ITS2 sequences and their respective secondary structures. This analysis indicates at least two distinct species in Trichoplax (Placozoa) supporting a recently suggested hypothesis, that Placozoa is “no longer a phylum of one”.

The paper includes data on rare occurring pseudofilamentous freshwater red alga, Chroodactylon ornatum (C. Agardh) Basson, in Poland and Slovakia, Central Europe. The species is classified as widespread in the whole world in littoral of stagnant waters or in rivers but rarely noted. Original microphotographs and drawings of Ch. ornatum are presented.

Total sixteen bacterial strains were isolated and purified from the samples collected from sugarcane molasses soil, sewage water and long-chain-hydrocarbon-contaminated area of the Punjab University, Lahore, Pakistan. Tolerance to different antibiotics was studied and strains showed multiple antibiotic resistance. All strains were characterized for Gram stain, biochemical reactions and polyhydroxyalkanoate (PHA) production. Total fourteen strains were Gram negative and two were Gram positive, while biochemically nine PHA producers showed affiliation to Pseudomonas, Enterobacter, Citrobacter, Bacillus and Escherichia. Screening for PHA production was done by Sudan black staining and nine out of sixteen strains exhibited PHA producing ability. PHA production was optimized for different growth parameters, like nitrogen concentration, pH and temperature. PHA extraction was done by solvent extraction method. Bacterial strains US1 and M1 accumulated up to 30% PHA of their cell dry weight on PHA extraction by solvent extraction method. Bacterial strain US1 was identified by 16S rRNA gene analysis as P. aeruginosa (DQ455691). PHA production was confirmed by PCR amplification of 500 bp fragment from PHA polymerase (Pha C) gene; five strains from nine PHA producers gave positive results on PCR. Pha C gene fragment of US1 was sequenced and submitted to Gene Bank under the accession number DQ455690. The amino acid sequence showed homology using the protein BLAST at 129–132 sites with different PHA synthases of the Pseudomonas sp.

The study presents habitat and phytosociological analyses of the Chara intermedia phytocenoses, rare described in Europe. 16 physico-chemical water parameters were analysed, coming from the samples taken in 20 phytocenoses of 13 lakes located in western Poland. The analysed community appeared in naturally shallow lakes representing last stages of the disappearance of glacial water basins. The study attempts to estimate the bioindicative value of the charophyta meadow Charetum intermediae in relation to its habitat. A particular attention has been paid to the determination of the habitat trophic condition, and to the concentration of elements connected with the hardness of water and the content of humic substances. The study shows crucial habitat gradients of the C. intermediae association, taking into account also the species composition of phytocenoses.

During vegetative period 2004–2005 powdery mildew (Erysiphe graminis DC. f. sp. hordei Em. Marchal) field resistance of spring barley cultivars was investigated at the Lithuanian Institute of Agriculture. The spring barley genotypes tested were Lithuania-registered cultivars, cultivars from genetic resources collection, and the new cultivars used for initial breeding. In total, 23 resistance genes were present in the 84 cultivars studied. Among mono-genes only mlo and 1-B-53 showed very high resistance. Slight powdery mildew necroses (up to 3 scores) formed on cultivars possessing these genes. The maximal powdery mildew (PM) severity reached a score of 8.5 and the area under disease progress curve (AUDPC) a value of 1216.8. The cultivars ‘Primus’, ‘Astoria’, ‘Power’, ‘Harrington’ and ‘Scarlett’ were the most resistant among the non mlo cultivars. Severity of PM on ‘Primus’ reached a score of 3.5 (3.0 of PM necrosis) in average, the other cultivars were diseased from 4.5 (3.0) to 5.0 (2.0). The AUDPC values for these cultivars except ‘Scarlett’ were the lowest (85.0–145.3) among the other cultivars. The highest contrast in development of the other leaf diseases was between highly resistant and susceptible to PM cultivar groups. The fast development of PM depressed development of the other diseases 4.7 times.

Spatial genotypical diversity of Sesleria albicans Kit. ex Schultes was studied in a dry grassland community by isozyme analysis. The aim was to identify the genetical individuals within the studied population and to assess the species’ clonal growth parameters. Vegetative mobility and branching intensity were measured in field for the sake of the correct interpretation of the results. Five isozyme systems were analyzed and interpreted (MDH, MNR, 6-PGDH, SkDH, LAP). Altogether, 98 distinct isozyme profiles were identified within an area of 2 m2. Average genotype identity rapidly decreased with distance. Several very remote ramets (more than 1 m) belonging to the same genet were identified. The longest distance between ramets of the same genotype was 153 cm. With average annual rhizome increment of 13.59 mm assessed for the studied population the age of genet with the most remote ramets exceeded 56 years by a bi-directional growth and 112 years by a growth in single direction. Number of daughter tillers produced by a tiller per year was 0.67 (branching intensity, median) and the median of tiller life span was 2.5 years. The high genotypical diversity of S. albicans in the studied population could be a result of both, regular and continuous seed production with subsequent seedling recruitment and long life span of genets.

Nine vegetation types were distinguished using cluster analysis within Molinion meadows in Slovakia. Vegetation of cluster 1 occurs on most acidic soils and is characterized by the occurrence of species of the Caricion fuscae alliance and of the Nardus grasslands. Vegetation of cluster 2 is also found on rather acidic soils but in contrast to cluster 1 vegetation it contains species of base-rich sites, such as Betonica officinalis, Galium boreale or Serratula tinctoria. Vegetation of cluster 3 occurs in wet base-rich habitats and often contains species of the Caricion davallianae alliance. Species of dry and Nardus grasslands are typical for vegetation of cluster 4, which is found at the driest sites and is confined to oligotrophic sandy soils. Vegetation of clusters 5 and 6 occurs on moist mesotrophic soils. Their species composition is quite similar, the main difference being that the former includes species-poor relevés and the latter includes species-rich relevés. Relevés of cluster 7 include species of dry grasslands and some ruderal species and represent degraded types of inundated floodplain meadows of the Deschampsion alliance. Vegetation of clusters 8 is characterized by species of the Phragmito-Magnocaricetea class and of the Deschampsion alliance, and occurs in wet nutrient-rich habitats. Vegetation of cluster 9, which usually develops from vegetation of cluster 8 due to decrease in the ground-water table, often contains species of dry grasslands and mesic meadows. Except for relevés of clusters 1 and 7, all others can be assigned to the Molinietum caeruleae Koch 1926 association. Cluster 1 corresponds to the Junco effusi-Molinietum caeruleae Tüxen 1954 association. Average Ellenberg indicator values for relevés, which were passively projected on the ordination biplot of detrended correspondence analysis, showed that the first ordination axis correlates with nutrients, soil base status and temperature, and second axis with moisture.

Morphological and micromorphological characters of the nutlets of ten Onosma species in Turkey were reported. Studied fruit surfaces show some variations. Based on the structure and patterns of epidermis cells of the nutlets, two principal types can be distinguished. In Type 1, the epidermal cells of the nutlet surface are formed in a reticulate pattern with varied sizes and shapes of the meshes. This type is the most common type in the studied species. Seven of the species observed belong to the reticulate type, i.e. O. bracteosum, O. mutabile, O. aleppicum, O. bornmuelleri, O. aucheranum, O. araraticum and O. tauricum. In Type 2, the epidermal cells of the nutlet surface are formed in an elongated type with varied sizes and shapes. Three of the observed species belong to this type, i.e. O. roussaei, O. isauricum and O. stenolobum. Significant taxonomic differences between species were determined on the surface of the nutlets.

A new rare cold-inducible (RCI) gene designated Cbrci35 was cloned from Capsella bursa-pastoris, an edible wild herb, using the rapid amplification of cDNA ends (RACE) method. The full-length cDNA of Cbrci35 (Database Accession No.: AY566573) was 1300 bp and contained a 978 bp ORF encoding a precursor of 326 amino acid residues with a 23 amino acids signal peptide. The predicted Cbrci35 protein contained a peroxidase active site and proximal heme-ligand signatures, an RGD cell attachment sequence motif and two leucine zipper pattern motifs. Bioinformatics analysis revealed that Cbrci35 has a high level of similarity with RCI genes from Arabidopsis thaliana and peroxidases genes from other plants. RT-PCR analysis revealed that Cbrci35 expressed only in root. A cold acclimation assay showed that Cbrci35 was expressed immediately after cold triggering, but this expression was transient, suggesting that it concerns cold acclimation. But expression was not induced exposed to dehydration, salt stress or abscisic acid, indicating that it might be subjected specifically to cold regulation. These results indicate that Cbrci35 is an analogue of RCI genes and may participate in cold-response or increasing the freezing tolerance of plants.

Genetic variation in six natural populations of Scots pine (Pinus sylvestris L.) was determined with isoenzyme analyses. For this purpose, haploid female gametophytes of seeds and horizontal starch gel electrophoresis technique were used. A total of 17 loci and 58 alleles were observed in studying 10 enzyme systems. The average proportion of polymorphic loci for populations ranged from 58.8% to 70.6%. The average number of alleles per locus per population was 2.65. The mean estimated expected heterozygosity (He) of populations was 0.294. A rather high proportion of genetic diversity (96.4%) was due to within-population variation and the remaining (3.6%) was due to variation among populations. The level of gene flow (Nem) was found to be 6.69 per generation. Nei’s genetic distance coefficient ranged from 0.006 to 0.027 (mean 0.017) among all possible population pairs. The mean value of Nei’s genetic distance is similar to the values reported for other European Scots pine populations. The low mean value of Nei’s genetic distance among populations is enough to explain low interpopulation variation. According to genetic variation parameters, three out of six populations (Akdagmadeni-Yozgat, Refahiye-Erzincan and Vezirkopru-Samsun) appear to be preferable populations for genetic conservation and forest tree breeding programs.

Adventitious organogenetic structures were produced in vitro using cotyledon explants excised from 14 day-old pepper seedlings. The best response was observed on MS medium containing 5.7 μM indole-3-acetic acid and 8.8 μM 6-benzylaminopurine. However, when transferred onto elongation medium (MS + 2.8 μM gibberellic acid), these structures frequently developed into leaf-like features rather than into normal shoots. Interestingly, the histological study conducted on the cut end of the cotyledonary petiole revealed a direct induction of numerous teratological protuberances that arise around the cut end of the explant. On the contrary, typical organized bud meristems were rarely observed. Thus, the low number of plants obtained after transfer onto elongation medium seems to be a consequence of this teratological developmental process, frequently associated with fasciated and degenerative meristems, rather than a defect in shoot development from available meristems.

Tissue micronutrient content (B, Fe, Mn, Zn, Cu, Mo, Co) was compared in healthy and ill shoots of a rootless aquatic carnivorous plant Aldrovanda vesiculosa growing in an outdoor culture, in plants in the field, in different accessions from around the world, and in shoots and turions. On the basis of microelemental analyses, the observed disorder of shoot apices cannot be explained as deficiency of any of the above micronutrients.

The effect of low pH and aluminum on nitrogen uptake and metabolism was studied in roots of Lotus japonicus grown in hydroponic cultures. The low pH slightly suppressed root elongation, and this effect was accompanied by the suppression of nitrate and ammonia uptake, as well as the nitrate reductase activity. In spite of high resistance of young Lotus plants to short-term Al application, the one-day treatment of Al strongly reduced nitrate uptake and also the activity of nitrate reductase (NRA) in the apical parts of roots. The glutamine synthetase activity was also suppressed by Al treatment, but in lower extent. On the other hand, the ammonium uptake and nitrite reductase activity stayed unchanged by Al treatment and the values were practically the same as in control plants. These results support the view that nitrate uptake and nitrate reduction might be the main processes responsible for Al induced growth retardation in Lotus plants grown in mineral acid soils.

The morphology and morphometry of five soil haptorid ciliates (Apospathidium terricola, Diplites telmatobius, Armatoenchelys geleii, Paraenchelys terricola, and Spathidium aciculare), collected from a variety of terrestrial habitats in Slovakia (Central Europe), were investigated using live observation and protargol impregnation. Emphasis was given on the ciliary pattern, the fine structure of dorsal brush, as well as the shape and size of the extrusomes. The Slovak specimens of A. geleii differ considerably from other populations investigated in detail by body size and micronuclei number. Furthermore, the most important difference concerns the number of dorsal brush rows (4 vs. 3). Therefore, it was decided to establish a new subspecies, namely Armatoenchelys geleii barborae ssp. n., for the Slovak population. Shape and nuclear variants of A. terricola and P. terricola are presented and morphometrically characterized. Novel details on the dorsal brush structure of A. terricola and D. telmatobius are reported. Pair formation and exconjugant nuclear reconstruction in P. terricola are described. Two species, namely A. terricola and S. aciculare, are first records for the Holoarctic biogeographic region.

The genus Nepalomyia is the largest genus in the subfamily Peloropeodinae, with 47 described species. In the present paper the Nepalomyia pallipilosa species group is established for the following three described species and seven new species: Nepalomyia biseta sp. n., N. emeiensis sp. n., N. guangxiensis Zhang et Yang, N. liui sp. n., N. pallipilosa (Yang et Saigusa), N. ruiliensis Wang et Yang, N. sichuanensis sp. n., N. ventralis sp. n., N. yangi sp. n. and N. zengchengensis sp. n. Information on their distribution is also given or updated.

We examined the microclimate of different grasslands and the relationships between the seasonal changes in the microclimatic conditions and the structural characteristics of orthopteran assemblages in the most typical grassland types of the plain, hilly and submontaneous territories of Central Europe in 2000–2005 years. According to our results: (i) The seasonal changes of humidity were characteristic of the vegetation units; (ii) The microclimate components of grasslands with similar habitat structure could be similar independently from their ecological circumstances; (iii) The microclimatic changes influenced the structure of the orthopteran assemblages.

In the period 1993–2006, during investigation of reproduction biology of the bearded tit, 106 deserted nests of the species were collected in Slovakia, Austria and Italy and their arthropod fauna was analyzed. Occasionally introduced individuals of the pseudoscorpion Lamprochernes nodosus, a frequent species in Central Europe, were recorded in the nests. Altogether 984 individuals and 33 species of mesostigmatic mites (Acari) were found in 46.2% of the nests examined. The ectoparasite Ornithonyssus sylviarum was most abundant and frequent; it represented almost 68.3% of all individuals. Due to it, the parasitic mites predominated (69.4% of individuals). Other ecological groups were less represented: edaphic detriticols − 11.6%, coprophils − 10.7%, species of vegetation stratum − 8.2%, and nidicols − 0.2%. Beetles (40 species, 246 individuals) were present in 57 nests. Most of the beetles were strongly hygrophilous species inhabiting soil surface in the reed stands or other types of wetlands and the shore vegetation. Predators represented 59% of all individuals. They might find food in the nests, but none of the species had a close relationship to bird nests and represented 35% of species. All beetle species penetrated the nests occasionally, when ascending on the vegetation or searching cover during periods of increased water level. Occasionally, larvae and nymphs of the Dermacentor marginatus tick were found. They were most probably introduced by insectivores of the genus Neomys. Only one species of fleas, Ceratophyllus garei — a parasite of birds nesting in humid environment, was recorded in the nests.

Experimental studies of regeneration and repair processes of schistomelic walking legs and pedipalps in the early postembryonal stages of Tegenaria atrica were carried out in two parts. In the first part changes in the structure of appendages were produced by exposing the developing embryos to alternating temperatures (14 and 32°C, changing every 12 hours) as teratogenic factor. In the second, principal, part of studies after amputating a given fragment of a larval schistomelic appendage the appearance, then the growth and development of the external regenerates were observed. The completion of epimorphosis of the pedipalps followed at nymph IV stage (after four post-larval moults) that of the walking appendages in most cases at nymph V stage (after five post-larval moults). In control spiders repair processes were noted, which proceeded at a rate comparable to regeneration processes.

The morphology of the spermatheca and eggs of Odontotarsus purpureolineatus were studied by optical microscopy and scanning electron microscopy. The spherical eggs were about 1.35 mm long and 1.09 mm wide. The egg batches generally consist of 13–14 eggs. The egg surface is covered by polygonal (hexagonal and pentagonal shapes prevail) ridges and tiny chorionic tubercles. There were 8–10 aero-micropylar processes between the polygons. The spermatheca of O. purpureolineatus is characterized by a spherical spermathecal bulb, a pumping region, a flange of pump and dilation of spermathecal duct. Spermathecal processes and a median spermathecal dilation with sclerotized rod are missing. The spermathecal bulb and the pumping region possess many pores.

In addition to antibiotic properties, medicinal plants are important sources of chemicals with potential application as pesticides. The present study deals with antitermitic potential of seed extracts of Withania somnifera (Indian ginseng), Croton tiglium (jamalgoota) and Hygrophila auriculata (talimkhana). The seed extracts caused changes in tunneling behaviour, number of bacterial colonies in hindgut and activities of enzymes in midgut of Odontotermes obesus. C. tiglium showed the lowest LT50 (12.85 and 2.65 h) among the three seed extracts at concentrations of 50% (half dilution of the extract) and 100% (extract without dilution), respectively. There was no tunneling in soil treated with 100% concentration of seed extracts of W. somnifera and C. tiglium. Numbers of bacterial colonies in the gut of termites from soils treated with 50% and 100% concentrations of the three plants did not differ significantly, but they differed from those in termites from untreated soil. At 50% concentrations of seed extracts of the tested plants, the difference in hindgut enzyme activities was not obvious, however, at 100% concentrations the enzyme activities in the termites from soils treated with seed extracts significantly differed from controls and differences were also recorded between the plants.

Total lipid and the fatty acid compositions of phospholipid and triacylglycerol fractions, prepared from eggs, 3rd instars of larvae, pupae, male and female adults of Lertha sheppardi, were analyzed by gas chromatography and gas chromatography-mass spectrometry. The effect of diet (adults’ nutrition) on fatty acid composition of L. sheppardi adults was also investigated. Total lipid of L. sheppardi considerably increased in adults compared with immature stages. There was a significant decrease in total lipid level in larval stage in contrast with egg stage. Qualitative analysis revealed the presence of 14 fatty acids during all stages. The major components were C16 and C18 saturated and unsaturated components which are ubiquitous to most animal species. In addition to these components, one odd-chain (C17:0) and prostaglandin precursor fatty acids were found. The fatty acid profiles of phospholipids and triacylglycerols were substantially different. In phospholipid fraction, monounsaturated fatty acids were the major proportion of fatty acids in both sex of adults and pupae, whereas polyunsaturated fatty acids were the most dominant fatty acids in eggs and 3rd instars. Results of triacylglycerol fraction revealed that fatty acid composition of eggs had higher level of C16:1, C18:0 and C18:3n-3 content than that of 3rd instars and pupae, which suggests accumulation of energetic and structural reserve materials during embryonic development. At more advanced developmental stages, mainly in adult females, the amount of C16:1 increased once again, which may be related to the need for accumulation of sufficient energy and of carbon reservoir in the developing new vitellum. Percentages of C18:1 were significantly high in adult stages compared to other stages. These findings indicate that the accumulation and consumption of fatty acids fluctuate through different development stages. Diet did not effect the fatty acid composition of L. sheppardi adults.

Molecular weights of the skeletal muscle myosin, actin, troponin C and tropomyosin were compared in two frog species, Hyla japonica and Xenopus tropicalis, by SDS-PAGE and Western blot. Polyclonal antibody was produced using H. japonica skeletal muscle as the antigen. Polyclonal antibodies to nematode (Caenorhabditis elegans), mold slime (Physarum polycephalum), crab (Pagurus japonicus) and chicken skeletal muscle were also used. In H. japonica, the molecular weights of skeletal myosin, actin, troponin C and tropomyosin were 230, 42, 19 and 38 kDa, respectively, by using anti-C. elegans paramyosin, anti-P. polycephalum actin, anti-crab troponin C and anti-chicken gizzard tropomyosin antibodies. Molecular weights of the same proteins in X. tropicalis detected by the same antibodies were 230, 43, 20 and 40 kDa, respectively. In total, 29 protein bands were detected in H. japonica skeletal muscle and 24 bands in X. tropicalis by SDS-PAGE. The results revealed interspecific differences in molecular weights of selected skeletal muscle proteins and in the total skeletal muscle protein profiles between the two frog species.

In vitro proliferation and cytokine production were investigated in BALB/c mice splenic cell cultures that were stimulated with concanavalin A (ConA) and lipopolysaccharide (LPS) and simultaneously exposed to salivary gland extracts (SGE) of unfed and partially fed adult ixodid ticks (Ixodes ricinus, Rhipicephalus appendiculatus, Amblyomma variegatum). Generally, tick SGE enhanced proliferation of unstimulated splenocytes and SGE of unfed ticks suppressed mitogen induced proliferation. Partially fed R. appendiculatus and A. variegatum suppressed ConA responses, while partially fed I. ricinus stimulated both ConA and LPS induced proliferation. A. variegatum and R. appendiculatus females slightly enhanced LPS responses 2 days after attachment but suppressed them at the end of the slow feeding phase. In 72 h ConA induced cell cultures, interferon gamma (IFN-γ) production was suppressed by SGE of all ticks, interleukin (IL)-10 production was enhanced by unfed I. ricinus and partially fed A. variegatum males and IL-5 production was enhanced by feeding R. appendiculatus females and A. variegatum males. The study revealed variability in the responsiveness of murine splenocytes to SGE of different ixodid tick species, whereby patterns of host immunomodulation within one tick species differed between sexes and changed during feeding.

Studies indicate that leptin is involved in not only energy expenditure and food intake, but also in protection against apoptosis, in inflammation and in stimulation of proliferation in many cell types. However, leptin treatment increases the oxidative stress in many cell culture studies. This contradiction evoked a question of whether leptin acts as an oxidant or antioxidant on glial cells. We investigated the effect of leptin on glial cell survival and hydrogen peroxide (H2O2)-induced toxicity in vitro. The survival rate of the cells was determined by using 3-(4,5-D-methylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, thyazolyl blue (MTT) method. The cells obtained from the whole brain of 1–3 day-old rat were treated with 1, 10, 100 and 1000 ng/mL leptin for 24 or 72 h. Either the pretreatment of leptin alone for 5 h or leptin combined simultaneously with H2O2 or well known antioxidant glutathione (GSH) were applied to the cells. Malondialdehyde (MDA) levels were measured in cell lysates to which leptin was added for 24 h. The 100 and 1000 ng/mL leptin treatment for 72 h increased the glial viability by 19% and 36%, respectively. The dose of H2O2 that killed 75% of the cells was determined as 100 µM. GSH at different doses was applied as a positive control to the cells and the dose of 500 µM completely eliminated toxic effect of 100 µM H2O2. Either the pretreatment of leptin alone for 5 h or leptin combined simultaneously with H2O2 could not eliminate H2O2-caused toxicity. Furthermore, respective leptin doses did not change the glia MDA level. We suggest that leptin can increase glia survival dose dependently, but can not eliminate H2O2-induced oxidation in primary mixed glial cell culture.