Volume 69, Issue 2

Escalation in food industries unctuous wastes has led to serious anthropogenic problems to the environment. Parallel to “green strategy”, growing awareness in biological treatment emphasizes efficacy of enzymatic technology for bioremediation. Pertinently, researchers are in search for new lipase-lipid interaction for improved outcome. Rhodococcus species have documented inadequate evidences on lipase enzyme production. Consequent assessments on Rhodococcus isolates from Peninsular Malaysia have identified twelve promising strains as lipase producer. Interestingly, apart from usual lipolytic behaviour, Rhodococcus sp. exhibited significant level of lipase endogenously, while cryogenic grinding method effectively ruptured the cell. An isolate from petroleum-contaminated site, namely Rhodococcus UKMP-5M, projected the highest level of lipase specificity and has further been optimized. It was found out that the best specificity was apparent in acidic condition (pH 5) with 6% inoculum at 30°C for 72 hours of incubation. Due to high level of mycolic cell-surfactant developed in triacylglycerol supplements, cell lysis was employed with Triton X-100 detergent solubilisation. As a result, oil blend composed of various carbon-chain length fatty acids (composite 2) induces enzyme production extensively. Remarkably, R. UKMP-5M found to cater enzyme production without aid of inducer by nature, but additional carbon source like glucose represses lipase production. Further ability for biological treatment was revealed when the optimized R. UKMP-5M whole cell degraded waste cooking oil significantly by solubilizing fatty acids and commencing conversion into biomass. These qualities resemble practical new lipid-lipase biological lipid rich on-site treatment.

High quality genomic DNA is the first step in the development of DNA-based markers for fingerprinting and genetic diversity of crops, including mango (Mangifera indica L.), a woody perennial. Poor quality genomic DNA hinders the successful application of analytical DNA-based tools. Standard protocols for DNA extraction are not suitable for mango since the extracted genomic DNA often contains secondary metabolites that interfere with analytical applications. In this study, we employed an additional step to remove polysaccharides, polyphenols and secondary metabolites from genomic DNA extracted from young or mature leaf tissue; then a modified traditional cetyl trimethyl ammonium bromide (CTAB) method was applied. The use of 0.4 M glucose improved DNA quality and avoided contamination and browning by polyphenolics, relative to the traditional CTAB method. This is an easy and efficient method for genomic DNA extraction from both young and mature leaves of mango. The isolated DNA was free of polysaccharides, polyphenols, RNA and other major contaminants, as judged by its clear colour, its viscosity, A260/A280 ratio and suitability for PCR-based reactions. This modified protocol was also used to extract high quality genomic DNA from other woody perennials, including walnut, guava, lychee, pear, grape and sugarcane.

Endosymbiotic theory suggests that plastids originated from a photosynthetic bacterium that was engulfed by a primitive eukaryotic cell. In consequence, the chloroplast genome remains affected by this ancestral event, although it is reduced in size and the number of constituent genes. Most parts of the plastid genome have been transferred to the host cell nuclear genome and are nuclear-encoded. Thus, chloroplast proteins are synthesized in the cytosol as precursors with N-terminal extensions called transit peptides. The evolution of import machinery was required to transfer transit peptides to the stroma. Until the present, two protein complexes have been found to mediate the import process: the Toc (outer) and Tic (inner) envelope membrane translocons. The evolutionary origin of many Tic and Toc proteins has been established, but not for the Tic110 subunit. Tic110 binds signal peptides and serves as a scaffold for the recruitment of stromal components. In this study, we analyzed hydrophobic clusters, protein folds, and protein structure homology and we conclude that Tic110 is composed of fourteen repeated motifs related to HEAT-repeats. The explanation for the presence of such repeats in Tic110 is that membrane arrangement is found in separate domains and their probable function in the chloroplast import process is discussed.

Thioredoxins (Trxs) are small ubiquitous oxidoreductases that participate in dithiol-disulphide exchange reactions. They are present in all organisms from prokaryotes to higher eukaryotes and involved in a variety of cellular processes in higher plants. The expression pattern of the multigenic family of grape Trxsh was analysed in different tissues by semiquantitative RT-PCR. The grape Trxh genes were approximately expressed in all tissues, but with different expression patterns than each other. The in silico analysis of the promoter regions of grape Trxs h demonstrated the presence of a number of potential cis-acting elements to respond to environmental signals, suggesting that Trxsh may respond to a variety of environmental signals, including dehydration, salinity, heat, cold, heavy metals, light, pathogens, and plant hormones. The grape Trx h genes were also found to be differentially induced under abiotic stress conditions due to the presence of different putative regulatory elements in their promoters. Collectively, the distinct expression patterns and different responses to environmental stress conditions suggest that each isoform of the multigenic family of VvTrxh may have a specific and non-redundant function in grape.

A species of Taphrina was collected on Potentilla erecta in Slovakia for the first time. It causes yellowish thickening of stems and leaves. The fungus was identified as Taphrina tormentillae, that is quite common on Potentilla erecta in the north Europe.

Strains of Trichophaea abundans (Ascomycota, Pezizales, Pyrenomycetaceae) were isolated from Haplic Cambisol, Haplic Cambisol (Eutric), Litic Leptosol (Sceletic), Haplic Umbrisol, and Fulvic Andosol soils in Slovakia and from Rendzic Leptosol, Chernozem, Cambisol, drilosphere, and feacal pelets of the earthworm Allolobophora hrabei collected in southern Moravia. The Slovak soils markedly differ in pH, from extremely acidic (pHH2O 3.9) to weakly alkaline (pHH2O 7.4). T. abundans appeared as a post-burn species on Haplic Cambisol, which experienced a wildfire after a windthrow event (November 2004). In Moravia T. abundans was also isolated from sites situated in southern slopes, from time and time they affected by fires. We also note the growth of T. abundans on the surface of Fulvic Andosol after gradual heating in the lab to temperatures reaching 105°C. These findings confirmed by a review of the literature, indicate that T. abundans belongs to the group of heat-resistant fungi appearing in soils regularly influenced by the fire or grass calcining.

The aim of this study was to assess the production of secondary metabolites of Alternaria species isolated from grape berries and determined the occurrence of Alternaria mycotoxins in vitro and in dried berries. Direct morphological examination on different media was used for identification and HPLC/MS method for quantification of Alternaria mycotoxins. We isolated A. alternata and A. tenuissima and randomly selected strains in vitro and in dried berries. In vitro we identified the production of Alternaria metabolites, included mycotoxins: alternariol, alternariol methylether, tenuazonic acid. Beside that one in dry berries were in measurable concentrations: macrosporin A, tentoxin and altenuete (I, II).

Effects of salinity on growth, protein content, proline, catalase and antioxidant enzyme activity in callus of three halophytes of the Thar Desert; Salsola baryosma, Trianthema triquetra and Zygophyllum simplex were evaluated. Callus tissues were cultured on Murashige and Skoog’s medium containing different concentrations of NaCl (50, 100 and 200 mM). Increase in dry weight and soluble proteins were observed in the callus exposed to lower salinity (50 and 100 mM NaCl) in all the three species, whereas on the medium containing 200 mM NaCl, significant decrease in these two growth parameters was recorded. Under the salinity stress maximum proline accumulation was found in S. baryosma with parallel increase in soluble sugars. Among the three species, T. triquetra callus showed maximum CAT activity with 50 and 100 mM NaCl treatment, whereas the enzyme activity decreased at 200 mM NaCl treatment in all three species. The antioxidant potential steadily elevated under salt treatment in all the above three species using 1, 1-diphenyl-2-picrylhydrazyl (DPPH) and ferric reducing antioxidant potential (FRAP) assay. Whereas, superoxide dismutase (SOD) quenching were recorded maximum at low (50 and 100 mM) concentrations in all the three species. However, T. triquetra callus showed maximum total phenolic content (TPC) 15 mg GAE g−1 with the elevated concentration of NaCl up to 200 mM, and S. baryosma callus showed lower TPC as compared to both species. A significant correlation between antioxidant capacity and TPC was observed indicating that phenolic compounds are the major contributors to the antioxidant potential in these halophyte species. FRAP and DPPH activity of Z. simplex showed maximum correlation (R = 0.992), as compared to other two species. We can conclude that all the three species exhibit a protection mechanism by sustaining growth parameters and antioxidant capacity. Due to high antioxidant property of all these species, the plant extracts may be included in nutraceutical formulations.

An efficient protocol for plant regeneration through somatic embryogenesis was established from in vivo leaf explants of Swertia chirayita, a critically endangered medicinal herb. The highest frequency (76%) of embryogenic callus was induced on Murashige & Skoog (MS) medium supplemented with 0.5 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.5 mg/L kinetin (Kn) from in vivo leaf explants. Globular somatic embryos were induced and further matured from such embryogenic calli by subsequent culture on the same medium. The highest number of somatic embryos (48.83 ± 4.6) was recovered from embryogenic calli derived from leaf explants after 6 weeks of culture. Synthetic seeds were produced by encapsulating of torpedo stage embryos in sodium alginate (4% W/V) gel, dropped into 100 mM calcium chloride (CaCl2 · 2H2O) solution. The synthetic seeds were germinated on MS medium. The highest frequency of synthetic seed germination (84%) was observed on MS medium supplemented with 1.0 mg/L BA and 0.5 mg/L NAA. Regenerants were successfully acclimatized under ex vitro condition. This is the first report on synthetic seed production of S. chirayita. Application of these protocols would be helpful in reducing stress in natural habitat, and in long-term storage of elite genotypes through synthetic seed production.

In the present study, we compared the response to salinity of three plants from Brittany coast with contrasted ecological status: Limonium latifolium (salt marshes), Matricaria maritima (beach tops and sand dunes) and Crambe maritima (fixed dunes). Under controlled glasshouse conditions, the growth of the three plants decreased with increasing external salinity. L. latifolium and C. maritima exhibited the highest and lowest resistance to severe salt stress (400 mM), respectively. M. maritima could be considered as an intermediate species, since it tolerated salinity up to 200 mM. The same observation could be made with sodium absorption and acuumulation in plant tissues, the most tolerant species (L. latifolium being the least Na accumulator. Hydrogen peroxide (H2O2) and malondialdehyde (MDA), commonly produced in conditions of stress, accumulated significantly in salt treated C. maritima and M. maritima while not in the tolerant L. latifolium. The latter used glutathione reductase to maintain constant H2O2 levels under salt stress while peroxidases were very low and ascorbate peroxidase did not respond to salinity stimulation. The medium tolerant halophyte M. maritima used peroxidases to protect from NaCl-induced H2O2, while the sensitive C. maritima failed to detoxify H2O2 despite a sharp increase in catalase activity. Results showed that the three coastal species differ in resistance to salinity. They also suggested that the level of plant resistance to salinity could be attributed to differing mechanisms to manage the accumulation of sodium and cope with the oxidative damages.

Upland fringes of the White Carpathians (Czech Republic) are known to support grasslands with the world’s highest local plant species richness. We investigated whether this unusually high plant richness has a parallel in snail communities, whether patterns of species composition of snail and plant communities in grasslands co-vary and how they are affected by local environment and landscape history. We compared plant and snail communities of dry to mesic grasslands in three neigh bouring regions: (1) hilly lowland of the Central Moravian Carpathians, (2) upland fringes and (3) upland of the White Carpathians. Both snail and plant communities exhibited a strong gradient in species composition associated with altitude, annual temperature and precipitation, soil calcium and pH. However, there was no correlation between local species richness of plants and snails in individual plots. The upland fringes of the White Carpathians were richest in snail species, probably due to intermediate environmental conditions, supporting the occurrence of species with contrasting environmental requirements. The highest local numbers of plant species were also recorded there, although differences among regions were not significant. The regional species richness of plants was also highest in the upland fringes, whereas that of snails was highest in the hilly lowland. Similarities in the diversity patterns of plants and snails among regions suggest the importance of regional factors for local richness, although local abiotic factors, which are partly correlated with the three regions, also influence local species composition and richness.

In January and in May 2012 three individuals (two adults and one nymph) from the genus Allocaeculus (family Caeculidae) were found in the area of the National nature reserve Devínska Kobyla near Sandberg (SW Slovakia). This mite family has not been recorded in Slovakia yet. Data about its nearest occurrence come from the Czech Republic and Austria. Based on the specimens, Allocaeculus sandbergensis sp. n. is described.

This study analyzed the size at sexual maturity and reproductive period of populations of Hepatus pudibundus in three bays on the northern coast of São Paulo, Brazil. Crabs were collected monthly and the bottom-water temperature was measured at each collection point. The animals were sexed, measured for carapace width (CW), and their gonadal stages were determined. A total of 8,674 specimens were collected (2,435 males and 6,239 females). Adult males showed the highest mean CW; the size at maturity for both sexes was 32.5 mm CW. Reproduction was continuous and peaked in spring and summer, because of the greater availability of plankton food for the larvae. This pattern is typical in tropical and subtropical regions, unlike the seasonal reproduction found in temperate regions. Reproductive activity of females was not significantly correlated with bottom-water temperatures. Immatures and individuals in all stages of gonadal development were found throughout the sampling period and at all depths, probably because the species completes its entire reproductive cycle in that area.

The subfamily Agathidinae was studied in parts of northern Iran. Six species were collected and identified. Among them, Cremnops desertor (L., 1758) is recorded for the first time from Iran. Lytopylus persicus Farahani & Talebi sp. n. is described and illustrated. A key to the West Palaearctic species of the genus Lytopylus Förster and an updated checklist of Iranian Agathidinae are provided.

Major histocompatibility complex (MHC) class II molecules play an important role in the immune response of vertebrates. In this paper, full-length MHC IIA cDNA was isolated from swamp eel (Monopterus albus) by rapid amplification of cDNA ends PCR. The genomic structure, molecular polymorphism, tissue distribution, and immune response of the MHC IIA gene to bacterial challenge were investigated. The full-length cDNA (GenBank accession No.: KC616308) is 1,509 bp in length including an 83 bp-long 5′ untranslated region (UTR) and a 709 bp-long 3′ UTR, which encoded a 238 amino acids protein. In the 2,339 bp-long MHC IIA genomic DNA, four exons and three introns were identified. Sequence comparison exhibited that the deduced amino acid sequence shared 27.1–66.3% identity with those of other species. Seven alleles were identified from five healthy individuals. Number of alleles per individual diversified from two to five. Five different 5′ UTR sequences and two different 3′ UTR sequences from one individual may infer the existence of five loci at least. Real-time quantitative PCR demonstrated that swamp eel MHC IIA transcripts were ubiquitously expressed in ten tissues, but the expression level was distinctly different. Significant changes were observed in liver, spleen, kidney and intestine after challenged with pathogenic bacteria Aeromonas hydrophilia.

Impacts of invasive alien plant species are threatening biodiversity worldwide and thus it is important to assess their effects on particular groups of organisms. However, such impacts were studied mostly in case of plant or invertebrate communities and our understanding the response of vertebrate species to plant invasions remains incomplete. To improve our knowledge in this respect, we studied bird communities in riparian vegetation along the rivers with different levels of Reynoutria spp. invasion in the Czech Republic. These findings will be interesting for basic ecology enhancing our knowledge of consequences of plant invasions, as well as for conservation practice. We surveyed understory bird species in 26 vegetation blocks along parts of three rivers running from the Beskydy Mountains in spring 2011. We used principal component analysis to assess vegetation structure of particular blocks and the first axis ordinated the blocks according to the degree of invasion by Reynoutria spp. Using generalized linear mixed-effects models we found that counts of Motacilla cinerea, Cinclus cinclus and Sylvia borin, as well as the total bird species richness, significantly decreased with increasing degree of Reynoutria spp. invasion, while Acrocephalus palustris showed the opposite pattern. These results suggest that Reynoutria spp. impacts negatively on the species strictly bond with river banks and habitats specialists, whereas habitat generalist species like Sylvia atricapilla were not affected. Preference of Acrocephalus palustris for Reynoutria spp. corroborates affinity of this species to large invasive herbs observed also in other studies. Our study showed that Reynoutria spp. invasion can reduce species richness of understory birds in riparian communities. Although the distribution of this plant species is still quite limited in central Europe, our results suggest that its more widespread occurrence could potentially threat some river bank bird species. Therefore, we urge for development of management actions that will act counter the Reynoutria spp. invasion.

The present work aimed to study the effect of palm pollen extract (PPE) as an anticoccidial and anti-apoptotic modulator during the course of murine intestinal Eimeria papillata infection. The fact that PPE has an anticoccidial efficacy against intestinal E. papillata infection in mice has been clarified by the reduction of faecal output of oocysts on day five post infection from 49.5 × 103 to 34 × 103 oocyst/g. Moreover, the number of intracellular eimerian stages of zygots and developing oocysts decreased by about 89% and that of schizonts and gamonts to 42% and 72%, respectively. E. papillata infection also induced an increase in the number of apoptotic cells from 17.5 to 122.8 apoptotic nuclei/10 villous crypt units (VCU). In addition, it caused a state of systemic inflammatory response as revealed by an elevation in levels of the pro-inflammatory biomarkers, inducible nitric oxide synthase (iNOs) and tumor necrosis factor alpha (TNF-α) from 5.3 and 78.3 to 33 pmol ml−1 and 96.3 pg ml−1 in blood, respectively, with concurrent duplication in the total leucocytic number. Upon treatment of infected mice with the aqueous PPE, the activity of iNOs was reduced by 55% and the level of TNF-α was decreased by 30%. Moreover, the total leucocytic count was significantly reduced from 9.05 × 103 to 7.8 × 103 cells/mm3. Based on our results, PPE showed both anti-coccidial, anti-inflammatory and anti-apoptotic activities. So it can be used in developing new herbal medicine against animal coccidiosis and may be suitable agent for treating eimeriosis associated inflammatory response.

The original version of the article was published in Biologia 68 (6): 1155–1163 (2013), DOI: 10.2478/s11756-013-0272-5. Unfortunately, the original version of this article contains a mistake in Table 1 and in left collumn, line 8 on page 1157. Here we display the corrected version of the table and text.

The original version of the article was published in Biologia 68 (6): 1198–1210 (2013), DOI: 10.2478/s11756-013-0268-1. Unfortunately, the original version of this article contains a mistake in Acknowledgements on page 1209. Here we display the corrected version of the Acknowledgements.