Volume 72, Issue 6

Discrimination study in opium poppy ( Papaver somniferum L.) genotypes has been performed by short tandem repeat (STR) markers derived from expressed sequence tags (ESTs) retrieved from the opium poppy DNA. Some of STRs originate from the gene sequences involved in the biosynthesis of morphine. Eight of twelve tested EST-derived STR markers generated polymorphism, 2-4 alleles were identified per locus, and the mean polymorphic information content value was 0.338. Seven cultivars developed in Slovakia, registered and released at present time for commercial cultivation were discriminated mutually. All the Slovak and Czech cultivars were also different from each other. EST-derived STR markers have shown a potential for discrimination of opium poppy genotypes cultivated for commercial production of either edible seeds or pharmacologically active alkaloids and could be applicable for the PCR-based forensic analyses.

Hybridization and polyploidization can be major mechanisms for plant evolution and speciation. Thus, the process of polyploidization and evolutionary history of polyploids is of widespread interest. The chloroplast DNA regions mat K was used to analyze to phylogenetic relationships and maternal donor of Elymus species and their closely related species. The Neighbor-Joining phylogenetic reconstructions partitioned the species into one monophyletic groups. All the Elymus species were related to species of Pseudoroegneria . These results indicated that Pseudoroegneria (St genome) was the maternal donor of the polyploidy Elymus . In addition, the St genome of Elymus had several origins and diverse species of Pseudoroegneria might have taken part in the formation of polyploid species of Elymus .

Blue light receptor phot1 and phot2 regulate hypocotyl phototropism induced by high intensity blue light (HBL) in functional redundancy. The phenotype of rpt2 single mutant is similar with double mutant phot1phot2 that lost phototropism, but rpt2phot1 double mutant shows normal phototropic response, which indicating that phot1 has dual opposite function with inducing and inhibiting hypocotyls phototropism. However, the mechanism of phot1-mediated inhibition remains to be elucidated. Here, we screened mutants with phototropism from EMS mutagenesis of rpt2 and cloned the gene P1SA1 ( P hototropin 1 S ignaling A ssociated 1 ), which maybe locate downstream of phot1 and function in regulating phot1-mediated inhibitory response. The rpt2 p1sa1 mutant shows normal phototropism to unilateral HBL and overexpression of PHOT1 in rpt2 p1sa1 lost hypocotyl phototropism again, which indicated that P1SA1 may be the allelic of PHOT1 . These results will open new perspectives about the screening and identification of genes involved in phot1-mediated inhibitory response, and provide theoretical basis for the self-regulation mechanism of plant to reduce high light damage.

The flower buds of Lonicera japonica are a rich source of antioxidants, comprising mostly flavonoids and phenolic acids, and in particular chlorogenic acid (5-caffeoylquinic acid, CGA), dicaffeoylquinic acid and caffeic acid. Hydroxycinnamoyl-CoA transferases are important for the synthesis of CGA. Here, the isolation and characterization of a Lonicera japonica gene encoding a hydroxycinnamoyl-CoA quinate transferase (HQT) is described. The recombinant LjHQT, produced by heterologously expressing the gene in Escherichia coli , favored p -coumaroyl-CoA over caffeoyl-CoA as its acyl donor, and expressed a higher affinity for quinate over shikimate as its its acyl acceptor. Southern blot analysis showed that the L . japonica genome harbors at least two copies of LjHQT . LjHQT transcript abundance was promoted both by spraying the leaves with salicylic acid or abscisic acid, and exposing them to a period of low temperature. The constitutive expression of LjHQT in Arabidopsis thaliana resulted in the production of CGA in the leaf and an higher level of resistance to the bacterial pathogen Pseudomonas syringae . CGA synthesis was positively correlated with LjHQT transcript abundance. When a fusion between the LjHQT promoter and GUS was expressed in A . thaliana , GUS expression was observed mainly in the sepal, vascular bundle, stigma and axillary bud.

Agriculture needs to be sustained by organic processes in current era as population explosion energy and the number of individuals undernourished are raising public concerns. Global warming poses additional threat by lifting the damage of salt stress especially in agro-economically vital crops like maize whose cultivation dates back to Mayans. To that end, cost-effective and organic fungal agents may be great candidates in stress resilience. We isolated the fungal strain from the soil of tea plants and characterized that via 5.8 S rDNA gene with internal transcribed spacer ITS-1 and ITS-2 regions, then named the target strain as TA. Reduced maximum quantum efficiency of PS II (Fv/Fm), the effective quantum yield of PS2 (ΦPS2), electron transport rate (ETR), photochemical quenching (qP) and increased non-photochemical quenching (NPQ) were detected in maize plants stressed with dose dependent salt. Enhanced Fv/Fm, ΦPS2, ETR, qP and decreased NPQ was observed in TA primed plus NaCl treated plants. TA biopriming significantly increased the lengths, fresh and dry weights of root/shoots and decreased the lipid peroxidation. Maize seedlings bioprimed with TA had less MDA and higher soluble protein, proline, total chlorophyll, carotenoid and RWC under NaCl. Furthermore, SOD, GPX and GR activities were much more increased in root and leaves of TA primed seedlings, however CAT activity did not significantly change. This is the first report to our knowledge that TA reverses the damage of NaCl stress on maize growth through improving water status, antioxidant machinery and especially photosynthetic capacity.

Genus Veigaia Oudemans, 1905 of the Veigaiidae family (Acari: Mesostigmata: Veigaiidae) is one of the most ubiquitous genera from Holarctic Region. In Romania 10 species were recorded as being distributed across the entire country. In the frame of this study we present the distribution data of all recorded Romanian Veigaia species: Veigaia cerva (Kramer, 1876); Veigaia exigua (Berlese, 1917); Veigaia kochi (Trägårdh, 1901); Veigaia nemorensis (C.L. Koch, 1839); Veigaia paradoxa Willmann, 1951; Veigaia planicola Berlese, 1892; Veigaia propinqua Willmann, 1936; Veigaia transisalae (Oudemans, 1902), Veigaia transylvanica Kontschán & Ujvári, 2008 and Veigaia uncata Farrier, 1957. At the same time, new characteristic ecological requirements for the Veigaia species were described (type of ecosystem, habitat, altitude and soil).

The main objective of this paper is to report the effect of shrub removal and mowing on the diversity and composition of ground-dwelling spider assemblages in Natura 2000 habitats of Mátra Mountains. We found significant effects of shrub removal and mowing on spider communities. Diversity decreased in the year following shrub removal but increased in the following years. Spider diversity in the final year decreased due to the lack of additional treatments. During our study the hay meadows were the most diverse habitats compared to control shrubs and treated shrubs. Treatments caused changes in community structure: the highest number of generalist species was observed in the treated shrubs, and the highest density of rare and protected species in the hay meadows. The high species turnover observed between hay meadows and control shrubs reflects the importance of grassland management. We conclude that shrub removal an effective grassland management action to increase spiders diversity in Natura 2000 habitats. Finally, treated shrubs require additional treatments such as mowing to ensure the spider communities inhabiting them are as diverse as those inhabiting meadows.

In spite of many investigations about the impact of reservoirs on river continuum, the downstream effects of a pumped-storage hydropower plants (PSHP) still have been less known. The aim of the study was to evaluate the downstream effect of the PSHP Čierny Váh in regard to the Serial Discontinuity Concept assumptions or studies assessing more frequent types of hydropower reservoirs. We predicted that the increase of water temperature and the alteration of food sources will hierarchically be the most relevant factors determining macroinvertebrate community composition. We focused on mayflies, caddisflies and non-insect benthic invertebrates due to their appropriate responses to discontinuity. We also determined the influence of the hydropower plant on life cycles and productions of the most abundant species of mayflies and caddisflies. We recorded the unique effect of the PSHP Čierny Váh on its downstream environmental variables. According to the nested ANOVA results, temperature regime, transported organic matter, benthic fine particulate organic matter and periphyton biomass were the environmental variables significantly modified by the river discontinuum. Significant differences were also found in the case of macroinvertebrate species richness, density and diversity. Nevertheless, based on the forward selection test for predictor effects, none of the environmental variables affected the macroinvertebrate assemblages significantly. However, the redundancy analysis revealed a separated group of macroinvertebrates preferring conditions characteristic for the site immediately below the reservoir. Regarding the impact of the reservoir on life histories, we revealed the shift in hatching, emergence and duration of life cycles of the abundant mayflies (except Baetis rhodani) and caddisfly Ecclisopteryx dalecarlica . Total annual production of mayflies as well as caddisflies was several times higher at the site immediately below the reservoir. Although PSHP Čierny Váh matches the SDC assumptions (except thermal stratification), the downstream impact resembles much more the effects of small reservoirs or epilimnetic-release reservoirs.

The parasite communities of three gudgeon species ( Gobio gobio, Romanogobio belingi and R. vladykovi ) were studied in two widely-separated rivers in the Czech Republic (Central Europe), the Bílina River (North Sea drainage) and Velička River (Black Sea drainage). Of the 25 parasite taxa identified, 21 species parasitised G. gobio , 8 species R. belingi and 13 species R. vladykovi . Due to the absence of specialist parasites, differences in the parasite community were mainly determined by the ecological conditions prevalent at the locality, which resulted in an increased similarity between sympatric species rather than between allopatric populations of the same species, i.e. G. gobio . Discriminant analysis showed significant differences between the four gudgeon populations. Five parasite species had significant power to discriminate particular parasite communities: the autogenic ectoparasites Gyrodactylus gobiensis , G. markakulensis and Paradiplozoon homoion ; the allogenic endoparasite Metorchis xanthosomus and the autogenic endoparasite Acanthocephalus anguillae . Only G. markakulensis occurred in both allopatric populations of G. gobio . Sympatric gudgeon populations were discriminated mainly by the presence of autogenic parasites, especially those with direct life cycles. Differences in parasite infection level between G. gobio and representatives of Romanogobio may reflect variance in diet and/or habitat use. Allopatric populations differed in the number of allogenic parasite species, which were more frequent in the Velička. Finally, the parasite community of R. belingi from the upper Elbe basin was characterised by a lack of gudgeon specific parasites, possibly supporting its non-indigenous status.

Karyotype and chromosomal characteristics of both minor and major rDNA of Sternopygus macrurus , a weakly electric South American fish, from two populations of the upper Paraná River basin, were investigated using conventional (Giemsa staining, silver staining, C-banding and base-specific fluorochromes) and molecular (fluorescent in situ hybridization (FISH) with 5S and 18S rDNA probes) cytogenetic techniques. Diploid chromosome number was invariably 2 n = 46 and karyotype composed of 23 pairs of biarmed chromosomes (28m+18sm). The nucleolus organizer regions (NORs) were located in the secondary constriction of the p arm of pair No. 2; this site corresponded with CMA 3 positive as well as with 18S rDNA signals, respectively. This 18S rDNA cluster was not syntenic to the 5S rDNA sites located at pairs Nos. 1, 5 and 15. The karyotypes and other chromosomal characteristics of individuals from the two populations in the upper Paraná River basin were identical. The karyotype differences among individuals identified as S . macrurus from Paraná River and the São Francisco and Amazon River basins, respectively, may indicate that these taxa might represent distinct species.

We used MaxEnt method as a tool to assess potential distribution for Rana pseudodalmatina Eiselt & Schmidtler, 1971 in regional scale. At the local scale of distribution, we evaluated patterns of sex-biased dispersal in breeding and non-breeding seasons. The MaxEnt model predicted potential suitable habitats for R . pseudodalmatina with high success rates (AUC Training data = 0.904 ± 0.057). According to the map constructed, the Iranian long legged wood frog prefers lowland forest regions, and land cover (65.4%); elevation (19.1%) has most impact on distribution of long-legged wood frogs in northern Iran. We found that, the percentage proportion of male frogs at the breeding site was significantly more than the percentage proportion of females (75.51% and 24.48%, respectively). Distance from the nearest potential breeding ponds differs between the sexes in breeding season and non-breeding season. It seems that, habitat destructions by local human activities and global climate change affected both regional and local distribution, are major threats to survival of the Iranian long-legged wood frog.

Rearrangement of the actin cytoskeleton plays an inductive role in chondrogenic differentiation. Our previous study showed that blebbistatin, an inhibitor of myosin II, removes actin stress fibres and induces chondrogenesis of mesenchymal cells in monolayer cultures. In the present study, we investigated signalling pathways implicated in the induction of chondrogenesis by dissolving actin stress fibres after blebbistatin treatment. Blebbistatin increased the activity of phosphoinositide 3-kinase (PI3K). Inhibition of PI3K with LY294002 blocked blebbistatin-induced chondrogenesis without affecting blebbistatin-induced reorganization of actin filaments. Blebbistatin also upregulated the phosphorylation of phosphoinositide-dependent protein kinase 1 (PDK1), and inhibition of PDK1 with GSK2334470 suppressed blebbistatin-induced chondrogenesis, indicating that removal of actin stress fibres by blebbistatin induced chondrogenesis by activating PI3K/PDK1. Although inhibition of Akt activity by Akt inhibitor IV blocked blebbistatin-induced chondrogenesis, phosphorylation of Akt was not affected by blebbistatin. Blebbistatin increased the phosphorylation of mammalian target of rapamycin (mTOR) at Ser2448 and p70 ribosomal protein S6 kinase (p70S6K). Inhibition of mTOR with rapamycin almost completely abolished the phosphorylation of p70S6K. Inhibition of mTOR complex 1 (mTORC1) and complex 2 (mTORC2) with pp242 diminished phosphorylation of Akt at Ser473, whereas inhibition of mTORC1 with rapamycin did not. However, blebbistatin did not affect the phosphorylation of mTOR at Ser2481. Taken together, the present results suggest that blebbistatin induces chondrogenesis by activating the PI3K/PDK1/mTOR/p70S6K pathway. Our data also indicate that Akt activity is essential for chondrogenesis but is regulated by mTORC2, which is independent of blebbistatin treatment.

Contractility of the heart muscle is a result of sliding movements between thick and thin filaments, produced by interactions between actin and myosin during the cross-bridge cycle. Activation of the myofilament is triggered by Ca 2+ binding to cardiac troponin C and is regulated through an “on/off” switching process occurring in the thin filament. Beside Ca 2+ regulation, strongly bound cross-bridges exert a positive feedback on myofilament regulation. Despite the importance of this positive feedback mechanism, its full molecular basis has so far remained elusive. Ca 2+ -regulated interactions between thick and thin filaments are widely regarded as an allosteric system, which means that multiple protein-protein interactions at their interface may exert alternative feedback effects on myofilament activation. To advance knowledge about these regulatory feedback mechanisms, we investigated a previously unstudied, hypothetical interaction between cardiac troponin and myosin, and how this interaction affects the function of myosin. Our results strongly suggest that myosin does indeed interact with the N-terminus of cardiac troponin I and the C-terminus of cardiac troponin T, suggesting a possible direct interaction between myosin and the IT-arm of troponin. We also conducted an in vitro heavy meromyosin (HMM) ATPase assay, and found that troponin significantly enhanced the actin-activated ATPase activity of HMM, both in the absence of tropomyosin and at the activated state of thin filament.