Pathways for the synthesis of polyesters in plants; cutin, suberin, and polyhydroxyalkanoates

Plants naturally produce the lipid-derived polyester cutin, which is found in the plant cuticle that is deposited at the outermost extracellular matrix of the epidermis covering nearly all aboveground tissues. Being at the interface between the cell and the external environment, cutin and the cuticle play important roles in the protection of plants from several stresses. A number of enzymes involved in the synthesis of cutin monomers have recently been identified, including several P450s and one acyl-CoA synthetase, thus representing the first steps toward the understanding of polyester formation and, potentially, polyester engineering to improve the tolerance of plants to stresses, such as drought, and for industrial applications. However, numerous processes underlying cutin synthesis, such as a controlled polymerization, still remain elusive. Suberin is a second polyester found in the extracellular matrix, most often synthesized in root tissues and during secondary growth. Similar to cutin, the function of suberin is to seal off the respective tissue to inhibit water loss and contribute to resistance to pathogen attack. Being the main constituent of cork, suberin is a plant polyester that has already been industrially exploited. Genetic engineering may be worth exploring in order to change the polyester properties for either different applications or to increase cork production in other species. Polyhydroxyalkanoates (PHAs) are attractive polyesters of 3-hydroxyacids because of their properties as bioplastics and elastomers. Although PHAs are naturally found in a wide variety of bacteria, biotechnology has aimed at producing these polymers in plants as a source of cheap and renewable biodegradable plastics. Synthesis of PHA containing various monomers has been demonstrated in the cytosol, plastids, and peroxisomes of plants. Several biochemical pathways have been modified in order to achieve this, including the isoprenoid pathway, the fatty acid biosynthetic pathway, and the fatty acid β-oxidation pathway. PHA synthesis has been demonstrated in a number of plants, including monocots and dicots, and up to 40% PHA per gram dry weight has been demonstrated in Arabidopsis thaliana. Despite some successes, production of PHA in crop plants remains a challenging project. PHA synthesis at high level in vegetative tissues, such as leaves, is associated with chlorosis and reduced growth. The challenge for the future is to succeed in synthesis of PHA copolymers with a narrow range of monomer compositions, at levels that do not compromise plant productivity. This goal will undoubtedly require a deeper understanding of plant biochemical pathways and how carbon fluxes through these pathways can be manipulated, areas where plant "omics" can bring very valuable contributions.

Geología del Montgrí

This paper, fundamentally stratigraphic, is based on the vertical distribution of fauna in sediments which constitute the Montgri Massif. The stratigraphic series is composed of two units: one allochtonous (Mesozoic) and a second one autochtonous (Paleogene). The latter crops out in front of the thrust belt and presents the typical facies described by PALLI (1972). The allochtonous part is formed by the following units (from bottom to top): variegated gypsiferous shales of Keuper age; marls, limestones and dolomites from the Lias (Domerian); gray breccia ted dolomites: supraliassic; and Cretaceous limestones with rare interbeds of calcareous marls and nodular shaly marls. In this unit the Neocomian, Barremian (?), Aptian, Albian, Cenomanian, Turonian (?) and Santonian haven been recognized. Structurally, the Montgri Massif is part of a thrust belt which has been divided into three parts of different development ang age. Both parts are affected by ample synclinal folds of NE-SW directions and fractures of NW-SE orientation. The fractures condition the torrential streams as well as the karstic phenomena present in the Massif. These and the eolian actions are the main causes of the present geomorphology of the Montgri

Managing Soybean Cyst Nematode, 2007

Soybean cyst nematode (SCN) causes the greatest yield loss of any single pathogen of soybean in Iowa. An estimated 50 million bushels were lost in Iowa to SCN in 2004. Damage from SCN is not limited to yield loss from root feeding; SCN also makes other diseases like sudden death syndrome, iron deficiency chlorosis, Pythium, Phytophthora root and stem rot and brown stem rot worse. Once established in a field, SCN cannot be eradicated. However, the use of multiple management tactics can help minimize yield loss.

8

Plants naturally produce the lipid-derived polyester cutin, which is found in the plant cuticle that is deposited at the outermost extracellular matrix of the epidermis covering nearly all aboveground tissues. Being at the interface between the cell and the external environment, cutin and the cuticle play important roles in the protection of plants from several stresses. A number of enzymes involved in the synthesis of cutin monomers have recently been identified, including several P450s and one acyl-CoA synthetase, thus representing the first steps toward the understanding of polyester formation and, potentially, polyester engineering to improve the tolerance of plants to stresses, such as drought, and for industrial applications. However, numerous processes underlying cutin synthesis, such as a controlled polymerization, still remain elusive. Suberin is a second polyester found in the extracellular matrix, most often synthesized in root tissues and during secondary growth. Similar to cutin, the function of suberin is to seal off the respective tissue to inhibit water loss and contribute to resistance to pathogen attack. Being the main constituent of cork, suberin is a plant polyester that has already been industrially exploited. Genetic engineering may be worth exploring in order to change the polyester properties for either different applications or to increase cork production in other species. Polyhydroxyalkanoates (PHAs) are attractive polyesters of 3-hydroxyacids because of their properties as bioplastics and elastomers. Although PHAs are naturally found in a wide variety of bacteria, biotechnology has aimed at producing these polymers in plants as a source of cheap and renewable biodegradable plastics. Synthesis of PHA containing various monomers has been demonstrated in the cytosol, plastids, and peroxisomes of plants. Several biochemical pathways have been modified in order to achieve this, including the isoprenoid pathway, the fatty acid biosynthetic pathway, and the fatty acid β-oxidation pathway. PHA synthesis has been demonstrated in a number of plants, including monocots and dicots, and up to 40% PHA per gram dry weight has been demonstrated in Arabidopsis thaliana. Despite some successes, production of PHA in crop plants remains a challenging project. PHA synthesis at high level in vegetative tissues, such as leaves, is associated with chlorosis and reduced growth. The challenge for the future is to succeed in synthesis of PHA copolymers with a narrow range of monomer compositions, at levels that do not compromise plant productivity. This goal will undoubtedly require a deeper understanding of plant biochemical pathways and how carbon fluxes through these pathways can be manipulated, areas where plant "omics" can bring very valuable contributions.

MAR elements regulate the probability of epigenetic switching between active and inactive gene expression.

Gene expression often cycles between active and inactive states in eukaryotes, yielding variable or noisy gene expression in the short-term, while slow epigenetic changes may lead to silencing or variegated expression. Understanding how cells control these effects will be of paramount importance to construct biological systems with predictable behaviours. Here we find that a human matrix attachment region (MAR) genetic element controls the stability and heritability of gene expression in cell populations. Mathematical modeling indicated that the MAR controls the probability of long-term transitions between active and inactive expression, thus reducing silencing effects and increasing the reactivation of silent genes. Single-cell short-terms assays revealed persistent expression and reduced expression noise in MAR-driven genes, while stochastic burst of expression occurred without this genetic element. The MAR thus confers a more deterministic behavior to an otherwise stochastic process, providing a means towards more reliable expression of engineered genetic systems.

Behavior of Eucalyptus grandis and E. cloeziana seedlings grown in arsenic-contaminated soil

Arsenic has been considered the most poisonous inorganic soil pollutant to living creatures. For this reason, the interest in phytoremediation species has been increasing in the last years. Particularly for the State of Minas Gerais, where areas of former mining activities are prone to the occurrence of acid drainage, the demand is great for suitable species to be used in the revegetation and "cleaning" of As-polluted areas. This study was carried out to evaluate the potential of seedlings of Eucalyptus grandis (Hill) Maiden and E. cloeziana F. Muell, for phytoremediation of As-polluted soils. Soil samples were incubated for a period of 15 days with different As (Na2HAsO4) doses (0, 50, 100, 200, and 400 mg dm-3). After 30 days of exposure the basal leaves of E. cloeziana plants exhibited purple spots with interveinal chlorosis, followed by necrosis and death of the apical bud at the 400 mg dm-3 dose. Increasing As doses in the soil reduced root and shoot dry matter, plant height and diameter in both species, although the reduction was more pronounced in E. cloeziana plants. In both species, As concentrations were highest in the root system; the highest root concentration was found in E. cloeziana plants (305.7 mg kg-1) resulting from a dose of 400 mg dm-3. The highest As accumulation was observed in E. grandis plants, which was confirmed as a species with potential for As phytoextraction, tending to accumulate As in the root system and stem.

Body, Disease and Treatment in a Changing World. Latin texts and contexts in ancient and medieval medicine (Proceedings of the ninth International Conference «Ancient Latin Medical Texts», Hulme Hall, University of Manchester, 5th-8th September 2007),

This volume reflects a variegated and fruitful dialogue between classical and medieval philologists and historians of science, philosophy, literature and language as well as of medicine - the diverse range of interests that the history of medicine in the Graeco-Roman world and the medieval West continues to stimulate and draw on. A recurrent theme is the transformation of medical knowledge in different languages, literary forms and cultural milieux. Several papers concern editorial work in progress on unpublished texts, available only in manuscript or early printed editions. Ce recueil met en dialogue des spécialistes des textes médicaux latins de l'Antiquité et du Moyen Âge. Certaines analyses adoptent une approche sociolinguistique, d'autres s'intéressent à des questions de transmission et de réception, d'autres enfin livrent des études sur le lexique médical. Mais toutes concourent à éclairer une histoire culturelle de la médecine qui s'inscrit dans un monde en mutation. With a preface by D. R. Langslow, and contributions by M. Baldin, J. P. Barragán Nieto, P. P. Conde Parrado, D. Crismani, M. Cronier, C. de la Rosa Cubo, A. Ferraces Rodríguez, K.-D. Fischer, P. Gaillard-Seux, A. García González, V. Gitton-Ripoll, G. Haverling, F. Le Blay, B. Maire, G. Marasco, A. I. Martín Ferreira, I. Mazzini, F. Messina, Ph. Mudry, V. Nutton, M. Pardon-Labonnelie, R. Passarella, M. J. Pérez Ibáñez, S. Sconocchia, A. M. Urso, M. E. Vázquez Buján, and H. von Staden.

Arsenic toxicity in Acacia mangium willd. and mimosa Caesalpiniaefolia benth. seedlings

Acacia mangium and Mimosa caesalpiniaefolia are fast-growing woody fabaceous species that might be suitable for phytoremediation of arsenic (As)-contaminated sites. To date, few studies on their tolerance to As toxicity have been published. Therefore, this study assessed As toxicity symptoms in A. mangium and M. caesalpiniaefolia seedlings under As stress in a greenhouse. Seedlings of Acacia mangium and M. caesalpiniaefolia were grown for 120 d in an Oxisol-sand mixture with 0, 50, 100, 200, and 400 mg kg-1 As, in four replications in four randomized blocks. The plants were assessed for visible toxicity symptoms, dry matter production, shoot/root ratio, root anatomy and As uptake. Analyses of variance and regression showed that the growth of A. mangium and M. caesalpiniaefolia was severely hindered by As, with a reduction in dry matter production of more than 80 % at the highest As rate. The root/shoot ratio increased with increasing As rates. At a rate of 400 mg kg-1 As, whitish chlorosis appeared on Mimosa caesalpiniaefolia seedlings. The root anatomy of both species was altered, resulting in cell collapse, death of root buds and accumulation of phenolic compounds. Arsenic concentration was several times greater in roots than in shoots, with more than 150 and 350 mg kg-1 in M. caesalpiniaefolia and A. mangium roots, respectively. These species could be suitable for phytostabilization of As-contaminated sites, but growth-stimulating measures should be used.

Groundwater fluctuations and footslope ferricrete soils in the humid tropical zone of southern Cameroon

This paper discusses the relationship between the differentiation of ferruginous accumulations and the variable water saturation of footslope soil patterns. An analysis of the slope morphology of a typical hill in the forest zone of southern Cameroon and a seasonal survey of the levels of groundwaters, springs and rivers were considered in relation to the petrology of different soil patterns. The study site is a tabular hillock whose slopes present a progressive development from steep to gentle slopes. The variable residence time of water within the soil, creating an alternation of reducing and oxidizing conditions, affects oil chemistry, structure and lateral extension of the soil patterns. The ferruginous soil patterns, being formed on the footslopes, gradually increase in extent with decreasing slope angle and the relative rise of the groundwater level. The steep footslopes, where groundwater has a shorter residence time, show a soft mottled clay pattern, restricted to the bottom part of the slope. The moderate footslopes exhibit a deep permanent and a temporary perched groundwater table. The latter, with its regular capillary fringe, contributes to more reducing conditions within isolated domains in the soil patterns, and thus to the alternation with oxidizing conditions, generating a continuous hard soil pattern (massive carapace). The more gently dipping footslopes exhibit groundwater levels near the surface and also a significant amplitude of groundwater fluctuation. Iron, previously accumulated in moderate footslope patterns, is reduced, remobilized, and leached. The soil patterns formed develop into a variegated carapace, more extended along the slope, containing less iron, but nevertheless more hardened, due to the important fluctuations of the groundwater table. These patterns are limited to the zone of groundwater fluctuation and deteriorate as the water fluctuation zone recedes. Copyright (c) 2005 John Wiley & Sons, Ltd.

The Permo-Triassic boundary in the Antalya Nappes (Western Taurides - Turkey)

The Antalya nappes (western Taurides-Turkey)*consist of several tectonic units which document the*Southern Neotethyan paleomargin from the Arabo-African*shallow shelf to the oceanic crust.*The Kerner Gorge Units (Upper Antalya nappes)*show a full stratigraphical succession from Ordovician*to Late Cretaceous. A carbonate platform regime*appeared during Late Permian times and existed up*to the early Middle Triassic. For detailed investigations*on the Permo-Triassic boundary, two lithostratigraphic*profiles have been selected: the Curuk*dag and the Kerner Gorge sections.*The main results presented in this paper are : 1) the Late Permian Pamucak Formation (Midian-*Dzhulfian) consists of calcareous algae-foraminiferae*bearing black limestones, locally rich in*brachiopods, crinoids and bryozoae : 2) this black limestone is overlain by an oolitic*grainstone. In the Curuk dag section, the oolitic horizon*is capped by a thin level of calcrete type; emersive*conditions are also inferred by a strong diagenetic*change within the oolitic deposit : 3) the first Early Triassic fossils, appearing*within or above the oolitic grainstone, are microforaminifera*and Pseudoclaraia wangi (late Griesbachian*in age). In the Curuk dag, a rich Early Triassic*microforaminifera association seems linked to a microbiallite*boundstone facies : 4) the overlying unfossiliferous lime mudstone,*the oolitical thick bedded grainstone, the variegated*marly limestone and the vermicular limestone facies*are present. not only in southern Turkey but seem*constant through ' the entire peri arabo-african platform.*Striking similarities appear between the studied*profiles and the Bellerophon-Werfen succession in the*Southern Alps..

Genetic linkage maps of 'Pêra' sweet orange and 'Cravo' mandarin with RAPD markers

The objective of this work was to construct linkage maps of 'Pêra' sweet orange [Citrus sinensis (L.) Osbeck] and 'Cravo' mandarin (Citrus reticulata Blanco) using RAPD markers and the pseudo-testcross strategy. The parents were chosen according to the resistance/susceptibility to citrus variegate chlorosis (CVC). The segregation of 176 markers was analyzed in 94 progeny of F1 hybrids, which were obtained from controlled crossings. The linkage map of 'Pêra' sweet orange had 117 markers defined by 12 linkage groups, which spanned 612.1 cM. Only six markers could not be linked to the linkage group and 48.7% of the markers showed segregation distortion. The linkage map of 'Cravo' mandarin had 51 markers defined by 12 linkage groups, which spanned 353.3 cM. Only two markers did not link to the groups and 15.7% showed segregation distortion. The construction of linkage maps is relevant to future mapping studies of the inheritance of CVC, citrus canker and leprosis.

RT-PCR and Dot Blot hybridization methods for a universal detection of tospoviruses

Transcriptase reverse - polymerase chain reaction (RT-PCR) and dot blot hybridization with digoxigenin-labeled probes were applied for the universal detection of Tospovirus species. The virus species tested were Tomato spotted wilt virus, Tomato chlorotic spot virus, Groundnut ringspot virus, Chrysanthemum stem necrosis virus, Impatiens necrotic spot virus, Zucchini lethal chlorosis virus, Iris yellow spot virus. Primers for PCR amplification were designed to match conserved regions of the tospovirus genome. RT-PCR using distinct primer combinations was unable to simultaneously amplify all tospovirus species and consistently failed to detect ZLCV and IYSV in total RNA extracts. However, all tospovirus species were detected by RT-PCR when viral RNA was used as template. RNA-specific PCR products were used as probes for dot hybridization. This assay with a M probe (directed to the G1/G2 gene) detected at low stringency conditions all Tospovirus species, except IYSV. At low stringency conditions, the L non-radioactive probe detected the seven Tospovirus species in a single assay. This method for broad spectrum detection can be potentially employed in quarantine services for indexing in vitro germplasm.

Detection and partial characterization of an isolate of Groundnut ringspot virus in Solanum sessiliflorum

The cubiu (Solanum sessiliflorum) fruit, originating in the Amazon basin, is commonly used in that region for food, medicine, and cosmetics. In an experimental culture of cubiu, in order to evaluate its adaptation to conditions in the Northern region of the state of Rio de Janeiro, it was observed plants with mosaic symptoms. A cubiu plant was collected and analyzed to identify the etiological agent. After mechanical passage through a local lesion host, a host range test was performed. The virus induced chlorotic local lesions in Chenopodium quinoa, necrotic local lesions in Gomphrena globosa, mosaic in S. sessiliflorum, leaf and stem necrosis in tomato (Lycopersicon esculentum) 'Rutgers', mosaic and leaf distortion in Datura stramonium and Physalis floridana, and necrotic local lesions followed by systemic necrosis and plant death in four Nicotiana species. Electron microscopic observations of ultra thin sections from infected cubiu leaves showed the presence of spheroidal, membrane-bound particles typical of tospovirus species. Analysis of the nucleocapsid protein from concentrated virus particles indicated the presence of a 28 kDa protein. RT-PCR was performed after total RNA extraction from infected IPA-6 tomato leaves. A fragment of approximately 0,8 kbp corresponding to the N gene was amplified, cloned and sequenced. The N protein from the cubiu isolate was 95% homologous to the Groundnut ringspot virus (GRSV) protein, and no more than 85% homologous to those from Zucchini lethal chlorosis virus (ZLCV) and Chrysanthemun stem necrosis virus (CSNV), Tomato spotted wilt virus (TSWV), and Tomato chlorotic spot virus (TCSV). This is the first report of the occurrence of GRSV (or any other plant virus) in cubiu.

Cyanobacteria from the Baltic Sea and Finnish lakes as an energy source and modulators of bioenergetic pathways

Cyanobacteria are a diverse group of oxygenic photosynthetic bacteria that inhabit in a wide range of environments. They are versatile and multifaceted organisms with great possibilities for different biotechnological applications. For example, cyanobacteria produce molecular hydrogen (H2), which is one of the most important alternatives for clean and sustainable energy. Apart from being beneficial, cyanobacteria also possess harmful characteristics and may become a source of threat to human health and other living organisms, as they are able to form surface blooms that are producing a variety of toxic or bioactive compounds. The University of Helsinki Culture Collection (UHCC) maintains around 1,000 cyanobacterial strains representing a large number of genera and species isolated from the Baltic Sea and Finnish lakes. The culture collection covers different life forms such as unicellular and filamentous, N2-fixing and non-N2-fixing strains, and planktonic and benthic cyanobacteria. In this thesis, the UHCC has been screened to identify potential strains for sustainable biohydrogen production and also for strains that produce compounds modifying the bioenergetic pathways of other cyanobacteria or terrestrial plants. Among the 400 cyanobacterial strains screened so far, ten were identified as high H2-producing strains. The enzyme systems involved in H2 metabolism of cyanobacteria were analyzed using the Southern hybridization approach. This revealed the presence of the enzyme nitrogenase in all strains tested, while none of them are likely to have contained alternative nitrogenases. All the strains tested, except for two Calothrix strains, XSPORK 36C and XSPORK 11A, were suggested to contain both uptake and bidirectional hydrogenases. Moreover, 55 methanol extracts of various cyanobacterial strains were screened to identify potent bioactive compounds affecting the photosynthetic apparatus of the model cyanobacterium, Synechocystis PCC 6803. The extract from Nostoc XPORK 14A was the only one that modified the photosynthetic machinery and dark respiration. The compound responsible for this effect was identified, purified, and named M22. M22 demonstrated a dual-action mechanism: production of reactive oxygen species (ROS) under illumination and an unknown mechanism that also prevailed in the dark. During summer, the Baltic Sea is occupied by toxic blooms of Nodularia spumigena (hereafter referred to as N. spumigena), which produces a hepatotoxin called nodularin. Long-term exposure of the terrestrial plant spinach to nodularin was studied. Such treatment resulted in inhibition of growth and chlorosis of the leaves. Moreover, the activity and amount of mitochondrial electron transfer complexes increased in the leaves exposed to nodularin-containing extract, indicating upregulation of respiratory reactions, whereas no marked changes were detected in the structure or function of the photosynthetic machinery. Nodularin-exposed plants suffered from oxidative stress, evidenced by oxidative modifications of various proteins. Plants initiated strategies to combat the stress by increasing the levels of alpha-tocopherol, mitochondrial alternative oxidase (AOX), and mitochondrial ascorbate peroxidase (mAPX).

Composite artistry in the Book of Numbers : a study in biblical narrative conventions

The present thesis discusses the coherence or lack of coherence in the book of Numbers, with special regard to its narrative features. The fragmented nature of Numbers is a well-known problem in research on the book, affecting how we approach and interpret it, but to date there has not been any thorough investigation of the narrative features of the work and how they might contribute to the coherence or the lack of coherence in the book. The discussion is pursued in light of narrative theory, and especially in connection to three parameters that are typically understood to be invoked in the interpretation of narratives: 1) a narrative paradigm, or ‘story,’ meaning events related to each other temporally, causally, and thematically, in a plot with a beginning, middle, and end; 2) discourse, being the expression plane of a narrative, or the devices that an author has at hand in constructing a narrative; 3) the situation or languagegame of the narrative, prototypical examples being factual reports, which seeks to depict a state of affairs, and storytelling narratives, driven by a demand for tellability. In view of these parameters the present thesis argues that it is reasonable to form four groups to describe the narrative material of Numbers: genuine narratives (e.g. Num 12), independent narrative sequences (e.g. Num 5:1-4), instrumental scenes and situations (e.g. Num 27:1-5), and narrative fragments (e.g. Num 18:1). These groups are mixed throughout with non-narrative materials. Seen together, however, the narrative features of these groups can be understood to create an attenuated narrative sequence from beginning to end in Numbers, where one thing happens after another. This sequence, termed the ‘larger story’ of Numbers, concerns the wandering of Israel from Sinai to Moab. Furthermore, the larger story has a fragmented plot. The end-point is fixed on the promised land, Israel prepares for the wandering towards it (Num 1-10), rebels against wandering and the promise and is sent back into the wilderness (Num 13-14), returns again after forty years (Num 21ff.), and prepares for conquering the land (Num 22-36). Finally, themes of the promised land, generational succession, and obedience-disobedience, operate in this larger story. Purity is also a significant theme in the book, albeit not connected to plot in the larger story. All in all, sequence, plot, and theme in the larger story of Numbers can be understood to bring some coherence to the book. However, neither aspect entirely subsumes the whole book, and the four groups of narrative materials can also be understood to underscore the incoherence of the work in differentiating its variegated narrative contents. Numbers should therefore be described as an anthology of different materials that are loosely connected through its narrative features in the larger story, with the aim of informing Israelite identity by depicting a certain period in the early history of the people.