Volume 70, Issue 7

The present study was conducted to isolate α-amylase-producing thermophilic bacteria from Darge¸cit hot springs in Turkey. The morphological, biochemical and physiological characterisation, as well as genetic analysis by 16S rRNA sequences indicated that the isolated strain AH1 was a member of Anoxybacillus genus. The strain was aerobe, Gram-positive and spore-forming rod, exhibiting optimum growth temperature and pH of 60ºC and 7.0-7.5, respectively. Optimization of growth medium and enzyme assay conditions for extracellular α-amylase production by the novel thermophilic Anoxybacillus sp. AH1 were carried out in many different media containing a variety of carbon and nitrogen sources. Among various carbon and nitrogen sources, peptone (2054.1 U/mL) at 1% and maltose (1862.9 U/mL) at 0.5% increased α-amylase activity, compared to controls. Moreover, a high enzyme production was observed with potato starch at 0.5% and 1% (2668.4 U/mL and 3627 U/mL, respectively), as well as with 1% soluble starch (2051.9 U/mL). The enzyme activity was found to be rather high in the presence of CaCl 2 up to 100 mM.

The chitinase producing bacterial strain was isolated from the vermicompost amended site of Mehsana district of Gujarat, India, and identified as Bacillus safensis MBCU6 using 16S rDNA sequencing. The chitinase was purified by ammonium sulfate precipitation followed by diethylaminoethanol sepharose CL-6B column chromatography. The purified enzyme could be demonstrated as a single band on sodium dodecyl sulfate polyacryalamide gel electrophoresis analysis as well as clear zone on zymogram, with estimated molecular mass of 58 kDa. The optimum pH and temperature of chitinase were pH 7.0 and 60◦C, respectively. The purified chitinase exhibited high degree of antifungal activity particularly against pathogenic Macrophomina phaseolina (60%) and Rhizoctonia solani (73%) by dissolving their cell wall components. The purified enzyme could hydrolyze colloidal chitin to its oligomers. It infers that the chitinase produced by Bacillus safensis may play a significant role in the activity as a biopesticide and bioactive material production.

Riparian zones support high biodiversity and are among the most vulnerable parts of the landscape. Their degradation facilitates the spread of invasive alien plant species. Our aim was to investigate the relationship between environmental characteristics of riparian zones and distribution and abundance of invasive alien plant taxa (IAT) along three rivers in Slovenia. In 100-m reaches of riparian zone we determined environmental characteristics and the presence and abundance of IAT. Canonical correspondence analysis was used to assess the relationship between IAT and environmental factors. The numbers and abundances of IAT in specific reaches were correlated with environmental factors using Spearman coefficient. These results revealed that the number and abundance of IAT in riparian zone were strongly positively correlated to the distance from river source, while they were negatively correlated with altitude, vegetation structure complexity, completeness of riparian zone and current velocity. Environmental factors that explained the distribution and abundance of IAT significantly were: the altitude, flow dynamics, distance from source, width of riparian zone, land-use, bank and channel structure, water depth and velocity. The research has pointed out the need for proper management of riparian zones to prevent invasion by IAT and also to strengthen other services provided by riparian zone as buffering anthropogenic impacts to water body, stabilising bank structure and increasing biodiversity.

The revision of the Paliurus spina-christi dominated vegetation of Europe is presented here. The study area includes the north-eastern part of Iberian Peninsula and Provence to the west, and spreads through the Apennine Peninsula to the Balkan Peninsula and up to Eastern Mediterranean areas. The phytosociological releves of Paliurus spina-christi dominated vegetation found in the literature for these European territories were gathered together in a phytosociological table. Floristic analysis, cluster analysis, and indirect gradient analysis were performed to determine the similarities and differences between the different Paliurus spina-christi dominated communities and their correlations with the main bioclimatic indices described in the literature. The various analyses highlight the existence of nine different associations (one of which is described here for the first time) that are attributed to different syntaxonomic levels. In the conclusion, a syntaxonomical scheme is proposed that classifies the European vegetation of shrublands in the class Rhamno-Prunetea which includes the class Paliuretea. Currently, we recognized three orders within this class: Prunetalia spinosae for central and south-central Europe; Pyro spinosae-Rubetalia ulmifolii within the Temperate oceanic bioclimate of the sub-Mediterranean variant; Paliuretalia spinae-christi with a central Mediterranean and sub-Mediterranean distribution, that mainly occurs in the central-eastern Mediterranean (Southern Apennine Peninsula) and the Balkans. Finally, a part of the shrub vegetation dominated by Paliurus spina-christi is referred to the class Quercetea ilicis, the order Pistacio lentisci-Rhamnetalia alaterni, the alliance Oleo sylvestris-Ceratonion siliquae and the suballiance Oleo sylvestris- Paliurenion spinae-christi that refers to thermophilous shrub communities that require high edaphic humidity.

In this work, potential contamination by invasive plant propagules as a result of trade exchange between Slovakia and Romania, was assessed. National lists, describing biology and ecology of 30 worst invasive plant taxa, were formulated, and trading in period 2006-2010 between countries analysed. Using norms for commodity impurity level, information on species habitat occupancy and literature data dealing with seed/fruit attachment on roads we calculated then potential invasive plant propagule export (PE) for each taxon. We found three fold higher total good export from Slovakia than in opposite direction, increasing export of commodities potentially containing invasive plant propagules exported from Romania to Slovakia and rise of road compared to railway transport. PEs for Slovak invasive plant taxa were one-two orders higher than those for Romanian ones. Potentially most exported taxa for Slovakia were: Amaranthus sp., Ambrosia artemisiifolia, Galinsoga sp., Kochia scoparia and Sorghum halepense (tens to hundreds tonnes each). And these could mostly be exported from Romania: Amaranthus sp., Ambrosia artemisiifolia, Artemisia annua, Conyza canadensis, Cuscuta campestris, Datura stramonium, Erigeron annus, Galinsoga sp., Iva xanthiifolia, Kochia scoparia, Lycium barbarum, Sorghum halepense, Veronica persica and Xanthium orientale subsp. italicum (units to tens tonnes each). High PE was significantly associated with cereals export. Our formula for PE is applicable for any inter- and intra-continental trade exchange.

Genetic transformation of plants offers the possibility of testing hypotheses on the function of individual genes and enables exploration of transgenes for targeted trait improvement. Cloning of the full-length class I chitinase from the Casuarina equisetifolia (CeChi1) was earlier reported by our team. In the present study, tobacco was used as a model system to functionally evaluate the potential of CeChi1 driven by ubiquitin promoter. The pUH-CeChi1 construct was introduced into tobacco by Agrobacterium-mediated transformation and the putative transformants were confirmed for stable gene integration, transgene expression and recombinant protein production using PCR, RT-qPCR, antifungal assays and in planta analysis. The in vitro antifungal bioassay using the total proteins from leaves of transformed plantlets revealed the characteristic lysis of hyphal tips of pathogenic fungi including Trichosporium vesiculosum, Fusarium oxysporum and Rhizoctonia solani. The in planta bioassay of transformed tobacco showed reduced disease symptoms when compared to untransformed wild plants. The study revealed that the class I chitinase isolated from C. equisetifolia can act as a potential gene resource in future transformation programs for incorporating disease tolerance.

The effects of drought and exogenous ABA on foliar anthocyanin accumulation and its role in drought tolerance were studied in two purple rice cultivars GS. No. 00621 and GS. No. 21427. The seedlings were grown for 14 days and separated into four groups including control (well-watered), drought (withholding water), ABA (foliar sprayed with 20 mg L−1 ABA and well-watered), ABA and drought (foliar sprayed with 20 mg L−1 ABA and withholding water). Drought was imposed by withholding water for 7 days. The results showed that drought significantly reduced root and shoot growth in GS. No. 00621, while had little effect on GS. No. 21427. Drought stress also induced anthocyanin overaccumulation in GS. No. 21427, whereas its content was reduced in GS. No. 00621. Exogenous ABA had no effect on anthocyanin in both wellwatered and drought conditions in both cultivars. Higher anthocyanin in GS. No. 21427 was coincided with higher DPPH scavenging ability, antioxidant enzyme activities as well as higher relative water content. It is, therefore, concluded that drought-induced anthocyanin overaccumulation may be different between purple rice cultivars and anthocyanin is beneficial for the response to drought stress by protecting plant antioxidant mechanism. Unlike anthocyanin in plant reproductive organs, exogenous ABA has no effect on foliar anthocyanin accumulation in purple rice. It is possible that anthocyanin accumulation in leaf and reproductive organs may be regulated differently or triggered by different mechanisms.

This study investigated the effects of sulphur foliar spray and S deprivation on Fe deficiency responses in two contrasting pea genotypes, Santi (tolerant) and Parafield (sensitive). Foliar application of sulphur enhanced morphological growth features, leaf chlorophyll score and root Fe chelate reductase activity predominantly in Santi and to a lesser extent in Parafield. These capacities eventually contribute to the higher Fe deficiency tolerance in Santi. These results are also important in terms of ameliorating Fe deficiency effects in peas through S foliar spray. Further, targeted investigation was performed on S deprivation in Santi and Parafield. S deprivation caused severe stunting, chlorosis and wrinkling in leaves and caused decrease in leaf Fe concentrations both in Santi and Parafield under Fe deficiency. S deprivation also led to a significant decrease in Fe chelate reductase and proton extrusion activities in both genotypes in Fe shortage. We conclude that S deficiency exacerbates Fe deficiency in peas by preventing the induction of the Fe chelate reductase activity and proton extrusion in roots. Taken together, these data confirm that Fe deficiency symptom expression and the Fe deficiency responses in peas are largely determined by S nutritional status.

Leptus (Leptus) delijanensis Khademi, Saboori & Hakimitabar sp. n. (Acari: Erythraeidae) is described and illustrated from larva ectoparasitic on undetermined grasshopper (Orthoptera: Acrididae) from Delijan, Markazi Province, Iran. It is the first report of Leptus species with solenidion of TFe I from Asia. A key to larval species of Leptus with solenidion on TFe I of the world is presented.

The aim of this study was to determine thrips abundance, species composition, sex ratio and seasonal dynamics of these insects on two morphological forms of Andean lupin (epigonal and semiepigonal). Thrips were studied in the lupine plants sown at two dates at the Experimental Research Station at Pawłowice, near Wrocław, Lower Silesia, Poland, during two growing seasons (2011-2012). The experiment was designed as a split-plot with four replicates for each morphological form and time of sowing. For the determination of thrips species’ composition, plants were collected from the central part of each plot when lupin was at the full blooming stage. In the laboratory, plants were shaken over a sheet of white paper. Once in each season thrips were also collected with a sweep net. In the laboratory larvae and adults were identified to species level and separated into males and females. During the whole lupin growing season, insects were also counted weekly on 10 consecutive plants in the middle row of each plot. Frankliniella intonsa, Thrips major and Aeolothrips intermedius were the dominant species in the full blooming stage of both semiepigonal and epigonal forms of Andean lupin. Thrips tabaci also occurred in greater numbers. There were no significant differences between numbers of thrips and number of species occurring on semiepigonal and epigonal plants sown at the same time. In the most numerous species, like F. intonsa and T. major, mainly females occurred in their population. In the population of A. intermedius males were abundant. Thrips occurring in Lupinus mutabilis, irrespective of the used form of lupin or sowing time, had one peak of abundance, i.e., the last ten days of June and the first ten days of July, when plants were at the blooming stage.

We estimated the seasonal dynamics and life cycle of the chironomid Heterotrissocladius marcidus (Walker, 1856) in three alpine lakes in the High Tatra Mountains (Central Europe). H. marcidus had univoltine cycle in all lakes surveyed and seasonal changes of density showed an early summer and an autumn peak interrupted by a mid-summer decline of larval abundance. Our results indicate that phenology of H. marcidus is sensitive to thermal regime and local differences in lake water temperature can lead to differences of the life history within the same population

The parasitic fauna in the lower Volga River basin was investigated for four gobiid species: the nonindigenous monkey goby Neogobius fluviatilis (Pallas, 1814), the round goby N. melanostomus (Pallas, 1814), the Caspian bighead goby Ponticola gorlap (Iljin, 1949), and the tubenose goby Proterorhinus cf. semipellucidus (Kessler, 1877). In total, 19 species of goby parasites were identified, of which two - Bothriocephalus opsariichthydis Yamaguti, 1934 and Nicolla skrjabini (Iwanitzki, 1928) - appeared to have been introduced from other geographic regions. The monkey goby had significantly fewer parasitic species (6), but relatively high levels of infection, in comparison to the native species. Parasitism of the Caspian bighead goby, which is the only predatory fish among the studied gobies, differed from the others according to the results of discriminant analysis. The parasitic fauna of the tubenose goby more closely resembled those of Caspian Sea gobiids, rather than the Black Sea monkey goby.

Although the external larval morphology of many species of Microhylidae has been characterized, ontogenetic studies in the family are scarce and partial. In the present study, we describe the development of the oral funnel, external gills, external nares, operculum, spiracle, and adhesive organs of Elachistocleis bicolor with the aim to contribute to future ontogenetic, evolutionary, and phylogenetic studies. The development of E. bicolor is similar to that of other species with free-swimming tadpoles as Microhyla ornata, Dermatonotus muelleri and Uperodon globulosus. The ontogeny of the oral apparatus shows only the early stages of the pattern most common in anurans. After the invagination of the stomodeum, the anterior and posterior labia are defined, but the keratinized structures are not formed. The conical adhesive glands are similar to those of Physalaemus and Dendropsophus tadpoles, but in E. bicolor, these develop in contact with the posterior labium. The external nares remain closed during most of the larval phase. During ontogeny, the external gills are simultaneously covered by a transverse operculum fold, which later advances towards the posteromedial ventral region, forming the spiracle, while in Type IV larvae the operculum folds close from right to left, covering the right gill first. The distinct larval characters of Microhylidae appear between the last embryonic stages and the first larval stages. Therefore, the consideration of these stages can be of great value in future studies of anuran development in phylogenetic and evolutionary contexts. Ontogenetic studies in microhylid tadpoles are fundamental to identify heterochronic processes, to interpret the variability of larval structures, and to establish or reinforce hypotheses about the origin of larval structures.

Strictly nocturnal or crepuscular habits on birds are restricted to a very limited number of species such as nightjars. Physiological skills of these species combined with ecological and behavioural adaptations provide them the ability to colonise a less competitive niche. Nightjars’ nocturnal ecological adaptations have been largely neglected on bird survey methods, which can bias the obtained results. Here, we studied the factors affecting vocal activity of two nightjar species, red-necked nightjar Caprimulgus ruficollis and European nightjar C. europaeus in a Mediterranean area of southern Portugal by modelling observed vocal activity of both species as a function of astronomic variables, particularly moon phase and day of the year. Red-necked nightjar vocal behaviour was positively associated with brighter moon phases and springtime, whereas European nightjar was only influenced by the season, being more active in the summer. This reveals the importance of light conditions on vocal activity of the former species. We suggest that specifically designed field surveys for these species should be preferentially conducted during brighter nights, when the lunar cycle is close to full moon. Furthermore, positive contacts with birds are maximized from late spring to early summer, at least for the red-necked nightjar.

A craniometric analysis of skulls of the wood mouse (Apodemus sylvaticus) from three different parts of Slovakia (north-western, northern and south-eastern) was conducted. Twenty-nine skull and dental variables were measured and evaluated on a sample of 56 adult individuals. The overlap of the values of the measured traits was high. Moreover, the differences among the populations were different for each trait. The non-parametric Kruskal-Wallis test and Discriminant function analysis showed variations in the cranial variables among the different parts of country. We have identified six morphometric variables suitable for the differentiation of populations on the basis of discriminant analysis. Significant differences were confirmed in the characteristics between the analyzed regions; thus, we can speak about small-scale craniometric differentiation. In addition, the results could indicate the influence of altitude, even over a shorter geographic distance between 48-49º N. Moreover, restriction of genetic interaction between the populations may in the future lead to even greater differences between the populations in Slovakia.

Methionine is the first amino acid residue in most of the recombinant proteins and sometimes it is necessary to remove the starting methionine in order to get foreign proteins with wild type sequences. Recombinant human interferon alpha-2b was produced in Escherichia coli in insoluble form as inclusion bodies, which were solubilized and refolded. Analysis of the purified recombinant protein indicated that it consisted of two types of molecules, one type, in majority, having N-terminal methionine intact, whereas the other type, in minority, having the N-terminal methionine cleaved by methionine aminopeptidase of the host. Both species were separated by high performance liquid chromatography and Nterminal methionine of the remaining molecules was removed by utilizing methionine aminopeptidase from Pyrococcus furiosus in order to get similar form of the interferon alpha-2b as produced by human body. The methodology developed in this study can be extended to other proteins of pharmaceutical importance.