A comparative study of Glycogen phosphorylase from selected cepbalopods

A comparative study of Glycogen phosphorylase from selected Cephalopods is reported in this thesis. A detailed investigation of an important glycolytic enzyme, phosphorylase, from a selected species, is undertaken. Loligo vulgaris, commonly known as squid, is selected as the source for the study. Phosphorylase is the key enzyme in the mobilization of chemical energy from glycogen and its role in the regulation of carbohydrate metabolism is well established. Although a good deal of information is available about phosphorylase from terrestrial animals, not much is known about the enzyme from aquatic fauna. In order to bridge this gap and also to compare the results with the findings from other sources, phosphorylase a was isolated from this marine mollusc and its properties studied in detail in this study

Das OEE-Protein 1 des Photosystem II (oxygen evolving enhancer) der Grünalge Scenedesmus obliquus besitzt Thioredoxin-Aktivität

Die Aminosäure-Sequenzierung an dem als "28 kDa-Thioredoxin f" beschriebenen Protein aus der Grünalge Scenedesmus obliquus hat gezeigt, dass dieses Protein mit dem als OEE bekannten Protein 1 aus dem Photosystem II identisch ist. Die früher postulierte Möglichkeit einer Fusion eines Thioredoxins mit einem Protein unbekannter Natur oder Insertion eines Thioredoxinfragments mit der typischen -Trp-Cys-Gly-Pro-Cys-Sequenz in ein solches Protein hat sich nicht bestätigt. Durch Anwendung einer auf das 33 kDa OEE-Protein ausgerichteten Präparationsmethode konnte gezeigt werden, dass das "28 kDa-Trx f" tatsächlich in den Thylakoidmembranen lokalisiert ist. Das Protein kann so innerhalb eines Tages in hoher Reinheit aus den Thylakoidmembranfragmenten eines Algenrohhomogenats isoliert werden; dabei bleibt die Fähigkeit des OEE-Proteins das chloroplastidäre Enzym Fructosebisphosphatase (FbPase) zu stimulieren erhalten. Mit gleichen Methoden wurden die Grünalgen Chlorella vulgaris und Chlamydomonas reinhardtii auf außergewöhnliche Proteine mit Trx-f Aktivität untersucht. Die hitze- und säurestabile Proteinfraktion aus Chlorella vulgaris enthält ein Protein mit vergleichbarer Molmasse von 26 kDa, das ähnlich wie in Scenedesmus eine Stimulation der chloroplastidären Fructosebisphosphatase zeigt. In dem hitze- und säurestabilen Proteinextrakt aus Chlamydomonas reinhardtii wird solche Aktivität nicht beobachtet. Eine Probe des rekombinanten, homogenen OEE-Proteins aus Spinat wurde auf Stimulation der chloroplastidären FbPase und NADPH-abhängigen Malatdehydrogenase (MDH) untersucht. Das Spinat OEE-Protein 1 zeigt mit diesen Enzymen keine Aktivität. Da das OEE-Protein 1 in Scenedesmus starke FbPase-Stimulation zeigt, die anderen Scenedesmus-Thioredoxine mit Molmassen von 12 kDa (Trx I und II) jedoch hohe Aktivität mit der zellulären Ribonucleotidreduktase zeigen, wird postuliert, dass das OEE-Protein die Funktion des Trx-f in vivo ersetzt.

Protokoll der Gewässerschau in Kassel am 19. April 2005

Bei der Begehung von Olebach, Losse und Alte Losse wurden der ökomorphologische Zustand von Sohle, Böschung, Linienführung, Verzahnung von Wasser und Land und der Ufergehölze sowie die Landnutzung notiert. Einleitungen von Abwässern und die Ablagerung von Abfällen auf Böschung und Ufer wurde protokolliert. Wassertemperatur und elektrische Leitfähigkeit wurden an mehreren Punkten gemessen. Die Leitfähigkeitswerte werden vor dem Hintergrund der Geologie der Einzugsgebiete interpretiert und zur chemischen Indikation von diffusen und punktförmigen Einleitungen genutzt. Es werden Maßnahmen zur Renaturierung, Maßnahmen gegen die Ablagerung von Gartenabfällen auf Ufer und Böschung, Maßnahmen gegen die ackerbauliche Nutzung und Bebauung im Gewässerrandstreifen vorgeschlagen.

Effects of different tillage treatments on labile soil organic matter pools and stabilization processes

An improved understanding of soil organic carbon (Corg) dynamics in interaction with the mechanisms of soil structure formation is important in terms of sustainable agriculture and reduction of environmental costs of agricultural ecosystems. However, information on physical and chemical processes influencing formation and stabilization of water stable aggregates in association with Corg sequestration is scarce. Long term soil experiments are important in evaluating open questions about management induced effects on soil Corg dynamics in interaction with soil structure formation. The objectives of the present thesis were: (i) to determine the long term impacts of different tillage treatments on the interaction between macro aggregation (>250 µm) and light fraction (LF) distribution and on C sequestration in plots differing in soil texture and climatic conditions. (ii) to determine the impact of different tillage treatments on temporal changes in the size distribution of water stable aggregates and on macro aggregate turnover. (iii) to evaluate the macro aggregate rebuilding in soils with varying initial Corg contents, organic matter (OM) amendments and clay contents in a short term incubation experiment. Soil samples were taken in 0-5 cm, 5-25 cm and 25-40 cm depth from up to four commercially used fields located in arable loess regions of eastern and southern Germany after 18-25 years of different tillage treatments with almost identical experimental setups per site. At each site, one large field with spatially homogenous soil properties was divided into three plots. One of the following three tillage treatments was carried in each plot: (i) Conventional tillage (CT) with annual mouldboard ploughing to 25-30 cm (ii) mulch tillage (MT) with a cultivator or disc harrow 10-15 cm deep, and (iii) no tillage (NT) with direct drilling. The crop rotation at each site consisted of sugar beet (Beta vulgaris L.) - winter wheat (Triticum aestivum L.) - winter wheat. Crop residues were left on the field and crop management was carried out following the regional standards of agricultural practice. To investigate the above mentioned research objectives, three experiments were conducted: Experiment (i) was performed with soils sampled from four sites in April 2010 (wheat stand). Experiment (ii) was conducted with soils sampled from three sites in April 2010, September 2011 (after harvest or sugar beet stand), November 2011 (after tillage) and April 2012 (bare soil or wheat stand). An incubation study (experiment (iii)) was performed with soil sampled from one site in April 2010. Based on the aforementioned research objectives and experiments the main findings were: (i) Consistent results were found between the four long term tillage fields, varying in texture and climatic conditions. Correlation analysis of the yields of macro aggregate against the yields of free LF ( ≤1.8 g cm-3) and occluded LF, respectively, suggested that the effective litter translocation in higher soil depths and higher litter input under CT and MT compensated in the long term the higher physical impact by tillage equipment than under NT. The Corg stocks (kg Corg m−2) in 522 kg soil, based on the equivalent soil mass approach (CT: 0–40 cm, MT: 0–38 cm, NT: 0–36 cm) increased in the order CT (5.2) = NT (5.2) < MT (5.7). Significantly (p ≤ 0.05) highest Corg stocks under MT were probably a result of high crop yields in combination with reduced physical tillage impact and effective litter incorporation, resulting in a Corg sequestration rate of 31 g C-2 m-2 yr-1. (ii) Significantly higher yields of macro aggregates (g kg-2 soil) under NT (732-777) and MT (680-726) than under CT (542-631) were generally restricted to the 0-5 cm sampling depth for all sampling dates. Temporal changes on aggregate size distribution were only small and no tillage induced net effect was detectable. Thus, we assume that the physical impact by tillage equipment was only small or the impact was compensated by a higher soil mixing and effective litter translocation into higher soil depths under CT, which probably resulted in a high re aggregation. (iii) The short term incubation study showed that macro aggregate yields (g kg-2 soil) were higher after 28 days in soils receiving OM (121.4-363.0) than in the control soils (22.0-52.0), accompanied by higher contents of microbial biomass carbon and ergosterol. Highest soil respiration rates after OM amendments within the first three days of incubation indicated that macro aggregate formation is a fast process. Most of the rebuilt macro aggregates were formed within the first seven days of incubation (42-75%). Nevertheless, it was ongoing throughout the entire 28 days of incubation, which was indicated by higher soil respiration rates at the end of the incubation period in OM amended soils than in the control soils. At the same time, decreasing carbon contents within macro aggregates over time indicated that newly occluded OM within the rebuilt macro aggregates served as Corg source for microbial biomass. The different clay contents played only minor role in macro aggregate formation under the particular conditions of the incubation study. Overall, no net changes on macro aggregation were identified in the short term. Furthermore, no indications for an effective Corg sequestration on the long term under NT in comparison to CT were found. The interaction of soil disturbance, litter distribution and the fast re aggregation suggested that a distinct steady state per tillage treatment in terms of soil aggregation was established. However, continuous application of MT with a combination of reduced physical tillage impact and effective litter incorporation may offer some potential in improving the soil structure and may therefore prevent incorporated LF from rapid decomposition and result in a higher C sequestration on the long term.

Biological Routes to Gold Nanoplates

Much effort has been devoted to the synthesis of gold nanoparticles with different shapes, including the zero-dimensional nanospheres, one dimensional nanorods, and two-dimensional nanoplates. Compared to zero or one dimensional nanostructures, the synthesis of two-dimensional nanostructures in high yield has always been more involved, often requiring complex and time-consuming steps such as morphology transformation from the nanospheres, or the seeded growth process. Herein we report a high yield method for gold nanoplate synthesis using the extract of unicellular green alga Chlorella vulgaris, which can be carried out under ambient conditions. More than 90% of the total nanoparticle population is of the platelet morphology, surpassing the previously reported value of 45%. The control of the anisotropic growth of different planes; as well as the lateral size, has also been partially optimized.

Láser Excimer en Vitiligo. ¿Tratamiento efectivo y seguro para esta enfermedad? Revisión sistemática de la literatura

Introducción: El vitíligo es una enfermedad prevalente en nuestro medio con una prevalencia del 2% de la población mundial. Los síntomas de esta enfermedad son principalmente estéticos al manifestarse como máculas acrómicas, simétricas en las extremidades y en rostro donde genera la mayor estigmatización de los pacientes. Actualmente ningún tratamiento provee mejoría pronta y permanente de los síntomas. Objetivo: Determinar la efectividad del Láser Excimer 308 nm en el tratamiento del vitíligo por medio de una revisión sistemática de la literatura. Métodos: Búsqueda sistemática de ensayos clínicos y estudios cuasiexperimentales en las bases de datos más importantes acerca de la efectividad del Láser Excimer 308 nm en la repigmentación de los pacientes adultos con vitíligo. Se evaluó su calidad metodológica. Resultados: De 862 artículos encontrados se escogieron 40 artículos potenciales de los cuales dos fueron incluidos en esta revisión. El láser Excimer 308 nm como monoterapia presenta una pigmentación efectiva (≥50%) en 28.03% de las áreas tratadas, de los cuales 72.9% se localizaron en áreas sensibles a radiación ultravioleta y 27.02% en zonas no sensibles. Inicio de pigmentación a la sesión número 13 (un mes post inicio del tratamiento). El láser fue seguro y bien tolerado. Conclusión: La evidencia sugiere que el tratamiento con Láser Excimer 308 nm, como monoterapia, es una alternativa terapéutica para lograr repigmentación pronta de las máculas acrómicas del vitíligo en áreas sensibles a radiación ultravioleta. Deben considerarse estudios que evalúen combinaciones de fármacos y Láser en el tratamiento de vitíligo.

Nest Boxes Facilitate Local-Scale Conservation of Common Goldeneye (Bucephala clangula) and Bufflehead (Bucephala albeola) in Alberta, Canada

We tested the general predictions of increased use of nest boxes and positive trends in local populations of Common Goldeneye (Bucephala clangula) and Bufflehead (Bucephala albeola) following the large-scale provision of nest boxes in a study area of central Alberta over a 16-year period. Nest boxes were rapidly occupied, primarily by Common Goldeneye and Bufflehead, but also by European Starling (Sturnus vulgaris). After 5 years of deployment, occupancy of large boxes by Common Goldeneye was 82% to 90% and occupancy of small boxes by Bufflehead was 37% to 58%. Based on a single-stage cluster design, experimental closure of nest boxes resulted in significant reductions in numbers of broods and brood sizes produced by Common Goldeneye and Bufflehead. Occurrence and densities of Common Goldeneye and Bufflehead increased significantly across years following nest box deployment at the local scale, but not at the larger regional scale. Provision of nest boxes may represent a viable strategy for increasing breeding populations of these two waterfowl species on landscapes where large trees and natural cavities are uncommon but wetland density is high.

Off-road sampling reveals a different grassland bird community than roadside sampling: implications for survey design and estimates to guide conservation

Grassland bird species continue to decline steeply across North America. Road-based surveys such as the North American Breeding Bird Survey (BBS) are often used to estimate trends and population sizes and to build species distribution models for grassland birds, although roadside survey counts may introduce bias in estimates because of differences in habitats along roadsides and in off-road surveys. We tested for differences in land cover composition and in the avian community on 21 roadside-based survey routes and in an equal number of adjacent off-road walking routes in the grasslands of southern Alberta, Canada. Off-road routes (n = 225 point counts) had more native grassland and short shrubs and less fallow land and road area than the roadside routes (n = 225 point counts). Consequently, 17 of the 39 bird species differed between the two route types in frequency of occurrence and relative abundance, measured using an indicator species analysis. Six species, including five obligate grassland species, were more prevalent at off-road sites; they included four species listed under the Canadian federal Species At Risk Act or listed by the Committee on the Status of Endangered Wildlife in Canada: Sprague’s Pipit (Anthus spragueii), Baird’s Sparrow (Ammodramus bairdii), the Chestnut-collared Longspur (Calcarius ornatus), and McCown’s Longspur (Rhynchophanes mccownii). The six species were as much as four times more abundant on off-road sites. Species more prevalent along roadside routes included common species and those typical of farmland and other human-modified habitats, e.g., the European Starling (Sturnus vulgaris), the Black-billed Magpie (Pica hudsonia), and the House Sparrow (Passer domesticus). Differences in avian community composition between roadside and off-road surveys suggest that the use of BBS data when generating population estimates or distribution models may overestimate certain common species and underestimate others of conservation concern. Our results highlight the need to develop appropriate corrections for bias in estimates derived from roadside sampling, and the need to design surveys that sample bird communities across a more representative cross-section of the landscape, both near and far from roads. 

Stomatal guard cells are totipotent

It has been successfully demonstrated, using epidermis explants of sugar beet (Beta vulgaris L.), that stomatal guard cells retain full totipotent capacity. Despite having one of the highest degrees of morphological adaptation and a unique physiological specialization, it is possible to induce a re-expression of full (embryogenic) genetic potential in these cells in situ by reversing their highly differentiated nature to produce regenerated plants via a callus stage. The importance of these findings both to stomatal research and to our understanding of cytodifferentiation in plants is discussed.

A high efficiency technique for the generation of transgenic sugar beets from stomatal guard cells

An optimized protocol has been developed for the efficient and rapid genetic modification of sugar beet (Beta vulgaris L.). A polyethylene glycol-mediated DNA transformation technique could be applied to protoplast populations enriched specifically for a single totipotent cell type derived from stomatal guard cells, to achieve high transformation frequencies. Bialaphos resistance, conferred by the pat gene, produced a highly efficient selection system. The majority of plants were obtained within 8 to 9 weeks and were appropriate for plant breeding purposes. All were resistant to glufosinate-ammonium-based herbicides. Detailed genomic characterization has verified transgene integration, and progeny analysis showed Mendelian inheritance.

Plants growing on contaminated and brownfield sites appropriate for use in Organisation for Economic Co-operation and Development terrestrial plant growth test

The Organisation for Economic Co-operation and Development (OECD) Terrestrial plant test is often used for the ecological risk assessment of contaminated land. However, its origins in plant protection product testing mean that the species recommended in the OECD guidelines are unlikely to occur on contaminated land. Six alternative species were tested on contaminated soils from a former Zn smelter and a metal fragmentizer with elevated concentrations of Cd, Cu, Pb, and Zn. The response of the alternative species was compared to two species recommended by the OECD; Lolium perenne (perennial ryegrass) and Trifolium pratense (red clover). Urtica dioica (stinging nettle) and Poa annua (annual meadow-grass) had low emergence rates in the control soil so may be considered unsuitable. Festuca rubra (chewings fescue), Holcus lanatus (Yorkshire fog), Senecio vulgaris (common groundsel), and Verbascum thapsus (great mullein) offer good alternatives to the OECD species. In particular, H. lanatus and S. vulgaris were more sensitive to the soils with moderate concentrations of Cd, Cu, Pb, and Zn than the OECD species.

Quantitative response of the longevity of seed of twelve crops to temperature and moisture in hermetic storage

Seeds of 39 seed lots of a total of twelve different crops were stored hermetically in a wide range of air-dry environments (2-25% moisture content at 0-50 degrees C), viability assessed periodically, and the seed viability equation constants estimated. Within a species, estimates of the constants which quantify absolute longevity (K-E) and the relative effects on longevity of moisture content (C-W) and temperature (C-H and C-Q) did not differ (P >0.05 to P >0.25) among lots. Comparison among the 12 crops provided variant estimates of K-E and C-W (P< 0.01), but common values of C-H and C-Q (0.0322 and 0.000454, respectively, P >0.25). Maize (Zea mays) provided the greatest estimate of K-E (9.993, s.e.= 0.456), followed by sorghum (Sorghum bicolor) (9.381, s.e. 0.428), pearl millet (Pennisetum typhoides) (9.336, s.e.= 0.408), sugar beet (Beta vulgaris) (8.988, s.e.= 0.387), African rice (Oryza glaberrima) (8.786, s.e.= 0.484), wheat (Triticum aestivum) (8.498, s.e.= 0.431), foxtail millet (Setaria italica) (8.478, s.e.= 0.396), sugarcane (Saccharum sp.) (8.454, s.e.= 0.545), finger millet (Eleusine coracana) (8.288, s.e.= 0.392), kodo millet (Paspalum scrobiculatum) (8.138, s.e.= 0.418), rice (Oryza sativa) (8.096, s.e.= 0.416) and potato (Solanum tuberosum) (8.037, s.e.= 0.397). Similarly, estimates of C-W were ranked maize (5.993, s.e.= 0.392), pearl millet (5.540, s.e.= 0.348), sorghum (5.379, s.e.=0.365), potato (5.152, s.e.= 0.347), sugar beet (4.969, s.e.= 0.328), sugar cane (4.964, s.e.= 0.518), foxtail millet (4.829, s.e.= 0.339), wheat (4.836, s.e.= 0.366), African rice (4.727, s.e.= 0.416), kodo millet (4.435, s.e.= 0.360), finger millet (4.345, s.e.= 0.336) and rice (4.246, s.e.= 0.355). The application of these constants to long-term seed storage is discussed.

Macrophyte community structure and species occurrence in relation to environmental determinants in the ephemeral aquatic habitats of Gavdos, Greece

The aims of this study were to explore the environmental factors that determine the distribution of plant communities in temporary rock pools and provide a quantitative analysis of vegetation-environment relationships for five study sites on the island of Gavdos, southwest of Crete, Greece. Data from 99 rock pools were collected and analysed using Two-Way Indicator Species Analysis (TWINSPAN), Detrended Correspondence Analysis (DCA) and Canonical Correspondence Analysis (CCA) to identify the principal communities and environmental gradients that are linked to community distribution. A total of 46 species belonging to 21 families were recorded within the study area. The dominant families were Labiatae, Gramineae and Compositae while therophytes and chamaephytes were the most frequent life forms. The samples were classified into six community types using TWINSPAN, which were also corroborated by CCA analysis. The principal gradients for vegetation distribution, identified by CCA, were associated with water storage and water retention ability, as expressed by pool perimeter and water depth. Generalised Additive Models (GAMs) were employed to identify responses of four dominant rock pool species to water depth. The resulting species response curves showed niche differentiation in the cases of Callitriche pulchra and Tillaea vaillantii and revealed competition between Zannichellia pedunculata and Chara vulgaris. The use of classification in combination with ordination techniques resulted in a good discrimination between plant communities. Generalised Additive Models are a powerful tool in investigating species response curves to environmental gradients. The methodology adopted can be employed for improving baseline information on plant community ecology and distribution in Mediterranean ephemeral pools.

PGR forum: serving the crop wild relative user community

The Euro-Mediterranean region is an important centre for the diversity of crop wild relatives. Crops, such as oats (Avena sativa), sugar beet (Beta vulgaris), apple (Malus domestica), annual meadow grass (Festuca pratensis), white clover (Trifolium repens), arnica (Arnica montana), asparagus (Asparagus officinalis), lettuce (Lactuca sativa), and sage (Salvia officinalis) etc., all have wild relatives in the region. The European Community funded project, PGR Forum (www.pgrforum.org) is building an online information system to provide access to crop wild relative data to a broad user community; including plant breeders, protected area managers, policy-makers, conservationists, taxonomists and the wider public. The system will include data on uses, geographical distribution, biology, population and habitat information, threats (including IUCN Red List assessments) and conservation actions. This information is vital for the continued sustainable utilisation and conservation of crop wild relatives. Two major databases have been utilised as the backbone to a Euro-Mediterranean crop wild relative catalogue, which forms the core of the information system: Euro+Med PlantBase (www.euromed.org.uk) and Mansfeld’s World Database of Agricultural and Horticultural Crops (http://mansfeld.ipk-gatersleben.de). By matching the genera found within the two databases, a preliminary list of crop wild relatives has been produced. Around 20,000 of the 30,000+ species listed in Euro+Med PlantBase can be considered crop wild relatives, i.e. those species found within the same genus as a crop. The list is currently being refined by implementing a priority ranking system based on the degree of relatedness of taxa to the associated crop.

The use of bioluminescence for monitoring in planta growth dynamics of a Pseudomonas syringae plant pathogen

The use of bioluminescence was evaluated as a tool to study Pseudomonas syringae population dynamics in susceptible and resistant plant environments. Plasmid pGLITE, containing the luxCDABE genes from Photorhabdus luminescens, was introduced into Pseudomonas syringae pv. phaseolicola race 7 strain 1449B, a Gram-negative pathogen of bean (Phaseolus vulgaris). Bacteria recovered from plant tissue over a five-day period were enumerated by counting numbers of colony forming units and by measurement of bioluminescence. Direct measurement of bioluminescence from leaf disc homogenates consistently reflected bacterial growth as determined by viable counting, but also detected subtle effects of the plant resistance response on bacterial viability. This bioluminescence procedure enables real time measurement of bacterial metabolism and population dynamics in planta, obviates the need to carry out labour intensive and time consuming traditional enumeration techniques and provides a sensitive assay for studying plant effects on bacterial cells.