Changes of photosynthesis under blue light were examined in the ABA-overproducing 7B-1 mutant in tomato. Net photosynthetic rate (PN), stomatal conductance (gs), intrinsic water-use efficiency (WUEi) and chlorophyll (a+b) [Chl (a+b)] content in leaves of different insertion (1st, 4th and 9th ones) were measured in 5-, 7- and 9-week-old plants. PN, gs, and Chl (a+b) content were mostly similar in young leaves of 7B-1 and wild type (WT) plants. With the aging of leaves, a blue-light-induced increase in PN and gs to steady-state was delayed and steady-state values of PN and gs were lower in 7B-1 plants compared with WT. Steady-state values of WUEi were increased in 4th and 9th leaves of 7B-1 plants compared with WT. The results can be explained by the higher endogenous level of ABA in 7B-1 plants and their lower sensitivity to ABA in earlier growth stage., E. Ježilová ... [et al.]., and Obsahuje bibliografii
Dysfunction of mitochondria induced by ischemia is considered to be a key event triggering neuronal cell death after brain ischemia. Here we report the effect of ischemia-reperfusion on mitochondrial protein synthesis and activity of cytochrome c oxidase (EC 1.9.3.1, COX). By performing 4-vessel occlusion model of global brain ischemia, we have observed that 15 min of global ischemia led to the inhibition of COX subunit I (COXI) synthesis to 56 % of control. After 1, 3 and 24 h of reperfusion, COXI synthesis was inhibited to 46, 50 and 72 % of control, respectively. Depressed synthesis of COXI was not a result of either diminished transcription of COXI gene or increased proteolytic degradation of COXI, since both Northern hybridization and Western blotting did not show significant changes in COXI mRNA and protein level. Thus, ischemia-reperfusion affects directly mitochondrial translation machinery. In addition, ischemia in duration of 15 min and consequent 1, 3 and 24 h of reperfusion led to the inhibition of COX activity to 90.3, 80.3, 81.9 and 83.5 % of control, respectively. Based on our data, we suggest that inhibition of COX activity is rather caused by ischemia-induced modification of COX polypeptides than by inhibition of mitochondrial translation., P. Racay ... [et al.]., and Obsahuje seznam literatury
The European stag beetle, Lucanus cervus, is recognised as a flagship species for biodiversity conservation. Although the species is widely distributed across Europe declines have led to it being granted protected or endangered status in a number of countries and regarded as "near threatened" by the IUCN. The integration of genetic approaches into conservation efforts is urgently needed but has been impeded to date by the lack of appropriate genetic markers. To provide such a resource the development of the first microsatellite loci for stag beetle is described. Loci were identified using two methods (i) enriched library cloning (ELC) and (ii) Restriction enzyme Associated DNA sequencing (RAD-Seq). Inefficient microsatellite detection using the ELC method suggests that RAD-Seq, or other Next Generation Sequencing based methods, may ultimately be more cost effective for obtaining informative suites of markers for this and other coleopteran species. 18 loci were characterised by genotyping 42 UK specimens collected as prey leftover/roadkill. All loci produced unambiguous genotypes and were polymorphic. Though preliminary, estimates of genetic variability suggest UK populations may be genetically depauperate. The microsatellite loci represent a suite of genetic markers that can be applied to non-invasive population monitoring and numerous other areas of Lucanus conservation and evolutionary research., Niall J. McKeown, Deborah J. Harvey, Amy J. E. Healey, Ilze Skujina, Karen Cox, Alan C. Gange, Paul W. Shaw., and Obsahuje bibliografii
In this study, we report the development of a set of 15 polymorphic microsatellite markers for the box tree moth, Cydalima perspectalis (Walker), a highly invasive insect in Europe causing significant damage to natural and ornamental Buxus trees. The markers were characterized for four distant populations in both its native (China, two populations) and invasive ranges (Czech Republic and Turkey, one population each). The number of alleles ranged from 2 to 12. No marker significantly deviated from the Hardy-Weinberg equilibrium for all the populations sampled. These microsatellite markers are promising tools for further studies on the invasive pathways and dispersal pattern of the box tree moth in Europe., Audrey Bras, Laure Sauné, Alain Roques, Jérôme Rousselet, Marie-Anne Auger-Rozenberg., and Obsahuje bibliografii
This paper describes the characterization of a set of nine microsatellite markers for the pipe-organ mud-dauber wasp, Trypoxylon albitarse F. (Hymenoptera: Crabronidae), a common solitary wasp species in Brazil. Eight of the nine loci described were found to be polymorphic. The number of alleles ranged from three to 11. Observed and expected heterozygosity on the population level ranged from 0.135 to 0.891 and 0.177 to 0.865, respectively. Genotypic analyses revealed no deviations from Hardy-Weinberg equilibrium or linkage disequilibrium after sequential Bonferroni correction. Cross-species amplification was also tested for nine related species and positive amplifications were found in most of the assays. These markers will be useful in studies involving relatedness analysis and investigations of the population genetics of these wasps., Juliano Da Costa Almeida ... [et al.]., and Obsahuje seznam literatury
Here we describe 16 novel microsatellite loci in the genome of the dragonhead sap beetle Thymogethes norvegicus, which were identified using a next-generation sequencing approach. In 40 dragonhead sap beetles from a population in southern Norway, we found 2-4 alleles per locus. Expected heterozygosity ranged from 0.18-0.65. Three of the loci showed significant deviation from Hardy-Weinberg equilibrium. After correcting for multiple tests, one pairwise locus combination displayed significant linkage disequilibrium. These novel microsatellite loci will be useful for future population structure, genetic diversity and conservation genetic studies of the dragonhead sap beetle., Oddmund Kleven, Anders Endrestøl., and Obsahuje bibliografii
a1_Photosystem (PS) II particles retaining a high rate of O2 evolution were isolated from the mesophilic filamentous cyanobacterium, Spirulina platensis. To achieve high production of PSII complexes in the cells, irradiance from halogen incandescent lamps was used. Disruption of cells by vibration of glass beads proved to be the most suitable procedure for isolation of thylakoid membranes. The selectivity of detergents for PSII particle preparation rose in the order of Triton X-100 < decyl-β-D-glucopyranoside < dodecyldimethyl-aminooxide < n-heptyl-β-D-thioglucoside < N-dodecyl-N,N-dimethylammonio-3-propane sulphonate < n-octyl-β-thioglycoside < octylglucoside < n-dodecyl-β-D-maltoside. The last four detergents yielded extracts, from which pure PSII particles not contaminated by PSI complexes could be obtained by sucrose-gradient centrifugation (20-45%) at the 43% sucrose level. We assumed both the acceptor and donor sides of the isolated n-dodecyl-β-D-maltoside (DM) particles to be intact due to high oxygen production by DM particles [1,500 meq(e-) mol-1 (Chl) s-1] achieved in the presence of all artificial acceptors tested. The PSII particle fraction from the sucrose gradient was used with immobilized metal (Cu2+) affinity chromatography (IMAC) for the preparation of the PSII core complex. By washing the column with a MES buffer containing MgCl2 and CaCl2, the phycobiliproteins were stripped off. The PSII core complex was eluted in a buffer containing 1% DM, mannitol, MgCl2, NaCl, CaCl2, and ɛ-aminocaproic acid. SDS-PAGE of the core complex provided pure bands of D1 and D2 proteins and PsbO protein from thylakoid membrane, which were used to raise polyclonal antibodies in rabbits. These antibodies recognized D1 and D2 not only as monomers of 31 and 32 kDa proteins, but also as heterodimers of D1, D2 corresponding to the band of 66 kDa on SDS-PAGE. This was in contrast to antibodies of, a2_synthetic determinants, which reacted only with the monomers of D1 and D2 proteins. These negative reactions against heterodimers of D1, D2 supported the hypothesis that dimeric forms of PSII reaction centre proteins have a C-terminal sequence sterically protected against a reaction with specific antibodies., and E. Šetlíková ... [et al.].
Early light-induced proteins (ELIPs) are nuclear-encoded thylakoid proteins. In the present research, two full-length cDNAs (741 and 815 bp), encoding ELIPs (190 and 175 aa) and their genomic sequences, were isolated from tea leaves, and named CsELIP1 and CsELIP2, respectively. Both the deduced CsELIPs contain a chloroplast transit peptide in the N-terminus and a chlorophyll a/b binding protein motif with three transmembrane helices in the C-terminus. The genomic sequences of the two CsELIPs conform to the three-exon pattern of ELIP genomic sequences of other plant species. However, the identities between two CsELIPs and ACJ09655 from gymnosperm species were higher than all of
ELIP-like proteins identified from other angiosperms. Expression analysis showed that the two CsELIP genes were significantly
up-regulated when the photoinhibition occurred in tea leaves, implying that they might be involved in photoprotection., X. W. Li ... [et al.]., and Obsahuje bibliografii
Sexual selection is thought to result in an elongation of genitalia in insects. Part of the genitalia of the leaf beetle Lema (Lema) coronata Baly is approximately twice the body length in both sexes. Nine microsatellite loci were isolated from L. (L.) coronata for use in future studies on the correlation between genital length and reproductive success. The nine loci were polymorphic, with three to six alleles per locus recorded in 40 individuals. Observed heterozygosity ranged from 0.08 to 0.38 and expected heterozygosity from 0.51 to 0.75. The non-exclusive probability of the second parent and the identity calculated from the set of loci were 0.011 and < 0.001, respectively. A deviation from the Hardy-Weinberg equilibrium was detected at all nine loci, i.e. significant heterozygote deficiencies, and several hypotheses accounting for this are discussed. The isolated loci carried independent information. Five polymorphic loci were amplified in a related sympatric species, L. (L.) scutellaris, which has relatively short genitalia. These microsatellite makers will be used to determine the paternity of offspring of females multiply mated in the laboratory and compare the selection pressures operating on genital length in these closely related species. and Yoko Matsumura, Izumi Yao.
A strain of Synechocystis sp. PCC 6803 expressing the yellow fluorescent protein (YFP) fused to the C-terminus of the PsaF subunit of PSI has been constructed and used to isolate native PSI complexes employing the GFP-Trap®, an efficient immunoprecipitation system which recognizes the green fluorescent protein (GFP) and its variants. The protein analysis and spectroscopic characterization of the preparation revealed an isolate of trimeric and monomeric PSI complexes, which showed minimal unspecific contamination as demonstrated by comparison with the wild type control. Interestingly, we detected CP43 subunits of PSII and small amounts of PSII core complexes specifically pulled-down with the YFP-PSI, supporting the association of PSII assembly modules and intermediate assembly complexes with PSI, as observed in our previous studies. The results demonstrate that the GFP-Trap® system represents an excellent tool for studies of PSI biogenesis and interconnection of PSI and PSII assembly processes., A. Strašková, J. Knoppová, J. Komenda., and Obsahuje bibliografické odkazy