Screening the toxicity and biodegradability of petroleum hydrocarbons by a rapid colorimetric method

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Expression studies in the embryo and in the micropylar endosperm of germinating coffee (Coffea arabica cv. Rubi) seeds

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Improvement to the physical quality and imbibition pattern in seeds of candeia (Eremanthus incanus (Less.) Less.)

The objective of this work was to develop a methodology for seed processing and X-ray analysis and to study imbibition pattern in seeds of candeia (Eremanthus incanus). Seeds were gathered in 2001 and 2002 in Morro do Pilar and Lavras respectively, processed and stored in a cold chamber (5ºC/60%RH) until the start of experiments. In order to identify unfilled seeds, a protocol was developed for X-ray use combining radiation energy (Kv) and exposure time (seconds). For elimination of unfilled seeds, an experiment was conducted using a South Dakota seed blower whereby different opening settings and ventilation times were combined. Original seed lots and categorized lots following radiograph viewing were tested by germination tests, with seeds being cleansed in sodium hypochlorite and scattered over blotting paper in 'gerboxes' and then taken to germinators with alternating temperatures of 20º-30ºC and 10 hours of light, for up to 15 days. Imbibition curves were determined under the conditions 30ºC, 20º-30ºC/10 hours of light and 30º-20ºC/10 hours of light. The X-ray protocol that best allowed viewing of internal seed structures was 30Kv for 45 seconds. Seed separation using the blower setting at a 3.0 opening for 30 seconds raised the number of embryonic seeds to values exceeding 99%, also raising the germination rates as a consequence. The imbibition pattern is three-phased and the germination process is completed in 72 hours. Under alternate temperatures, primary root growth is favored.

Phenolic compounds influence seed dormancy of palicourea rigida h.b.k. (rubiaceae), a medicinal plant of the brazilian savannah

Palicourea rigida H.B.K. (Rubiaceae), a medicinal species commonly known as douradinha, has wide distribution across ecosystems in Central and South America. This species exhibits seed dormancy delaying germination until optimal conditions for seedling growth and development are in place. While dormancy ensures species survival, it also presents a technical problem for developing P. rigida’s plant production program. Thus, the objective of this study was to investigate if secondary metabolites present in seeds influence the seed dormancy of P. rigida. Mature fruits were harvested from the native habitat, in the savanna region of the State of Minas Gerais during February 2009, 2010 and 2011. The content of phenolic compounds in the seed of P. rigida was measured, and the allelopathic effects were assessed using the germination of lettuces as model to detect phytotoxicity. The P. rigida seeds geminated at rates varying between 7% and 31% with a Seed Germination Index (SGI) of 0.09. Data suggest that the phenolic compounds present in the seeds may be responsible for seed dormancy.

Impacts of Plant Size, Density, Herbivory, and Desease on Native Platte Thistle (Cirsium Canescens)

Abstract. Based on prior field observations, we hypothesized that individual and interacting effects of plant size, density, insect herbivory, and especially fungal disease, influenced seedling and juvenile plant growth in native Platte thistle populations (Cirsium canescens Nutt.). We worked at Arapaho Prairie in the Nebraska Sandhills (May - August 2007), monitoring plant growth, insect damage, and fungal infection within different density thistle patches. In the main experiment, we sprayed half of test plants in different density patches with fungicide (Fungonil© Bonide, containing chlorothalonil) and half with a water control. Fungal infection rates were very low, so we found no difference in fungal attack between these treatments. However, plants that received the fungicide treatment had significantly faster growth over the season than did the control plants. At the same time, plants in the fungicide treatment had significantly reduced insect herbivory. These results strongly suggest that the fungicide had insecticidal effects and that insect herbivory significantly decreases juvenile Platte thistle growth. Further, damage by insect herbivores tended to be higher for larger plants, and herbivory was variable among different patches. However, plant density did not appear to have a large effect on the amount of insect herbivory that individual juvenile Platte thistle plants received. In the second experiment, we examined germination and survival success in relationship to seed density, and found that germination success was higher in areas of lower seed density. In the third experiment, we tested germination for filled seeds categorized primarily by color variation and size, and found no difference in germination related to either color or seed weight. We conclude that seed density, but not seed quality as estimated by color or size, affects germination success. Further, although herbivory was not significantly affected by plant density at any of the scales examined, insect herbivory significantly reduces the growth and success of juveniles of this characteristic native sand prairie plant.

A Novel Stress-Induced Sugarcane Gene Confers Tolerance to Drought, Salt and Oxidative Stress in Transgenic Tobacco Plants

Background: Drought is a major abiotic stress that affects crop productivity worldwide. Sugarcane can withstand periods of water scarcity during the final stage of culm maturation, during which sucrose accumulation occurs. Meanwhile, prolonged periods of drought can cause severe plant losses. Methodology/Principal Findings: In a previous study, we evaluated the transcriptome of drought-stressed plants to better understand sugarcane responses to drought. Among the up-regulated genes was Scdr1 (sugarcane drought-responsive 1). The aim of the research reported here was to characterize this gene. Scdr1 encodes a putative protein containing 248 amino acids with a large number of proline (19%) and cysteine (13%) residues. Phylogenetic analysis showed that ScDR1is in a clade with homologs from other monocotyledonous plants, separate from those of dicotyledonous plants. The expression of Scdr1 in different varieties of sugarcane plants has not shown a clear association with drought tolerance. Conclusions/Significance: The overexpression of Scdr1 in transgenic tobacco plants increased their tolerance to drought, salinity and oxidative stress, as demonstrated by increased photosynthesis, water content, biomass, germination rate, chlorophyll content and reduced accumulation of ROS. Physiological parameters, such as transpiration rate (E), net photosynthesis (A), stomatal conductance (gs) and internal leaf CO2 concentration, were less affected by abiotic stresses in transgenic Scdr1 plants compared with wild-type plants. Overall, our results indicated that Scdr1 conferred tolerance to multiple abiotic stresses, highlighting the potential of this gene for biotechnological applications.

Parasites in the neighbourhood: Interactions of the mistletoe Phoradendron affine (Viscaceae) with its dispersers and hosts in urban areas of Brazil

Mistletoes constitute an important food resource for animals in many ecosystems. However, these plants are considered pests in urban areas because of deleterious effects they have on the host trees. Studies in urban areas were mostly focused on listing host species or procedures to control the "pest". In this sense, broader studies including several aspects of mistletoes ecology in urban ecosystems are still missing. We studied the interaction of the mistletoe, Phoradendron affine, with its dispersers and hosts in two urban sites in Uberlandia, Brazil. Phoradendron affine fruits were consumed almost exclusively by Euphonia chlorotica, which was crucial for seed germination. Parasitism was recorded in five hosts, two native (Handroanthus chrysotrichus and Tabebuia roseoalba) and three exotic species (Spathodea campanulata, Ligustrum lucidum and Melia azedarach). Mistletoes were found parasitizing larger host trees, a trend commonly reported for mistletoe-host interaction. Mistletoe seed germination was not affected by the trees species, whether host or non-host, but the radicle of germinated seeds could not penetrate the bark and seedlings invariably died in non-host species. We found a high prevalence of parasitism in our study, in comparison to what previous studies reported for natural areas. The spatial distribution of the hosts and high light incidence on isolated host trees may lead to this high prevalence in urban areas. Rather than eradicated, mistletoes in urban areas should be ecologically managed and their importance for bird species conservation must be considered. More studies to determine which bird species are favoured by mistletoe presence in urban areas will be essential for, this purpose. (C) 2012 Elsevier GmbH. All rights reserved.

Adaptation and Constraint in the Plant Reproductive Phase

Conservatism is a central theme of organismic evolution. Related species share characteristics due to their common ancestry. Some concern have been raised among evolutionary biologists, whether such conservatism is an expression of natural selection or of a constrained ability to adapt. This thesis explores adaptations and constraints within the plant reproductive phase, particularly in relation to the evolution of fleshy fruit types (berries, drupes, etc.) and the seasonal timing of flowering and fruiting. The different studies were arranged along a hierarchy of scale, with general data sets sampled among seed plants at the global scale, through more specific analyses of character evolution within the genus Rhamnus s.l. L. (Rhamnaceae), to descriptive and experimental field studies in a local population of Frangula alnus (Rhamnaceae). Apart from the field study, this thesis is mainly based on comparative methods explicitly incorporating phylogenetic relationships. The comparative study of Rhamnus s.l. species included the reconstruction of phylogenetic hypotheses based on DNA sequences. Among geographically overlapping sister clades, biotic pollination was not correlated with higher species richness when compared to wind pollinated plants. Among woody plants, clades characterized by fleshy fruit types were more species rich than their dry-fruited sister clades, suggesting that the fleshy fruit is a key innovation in woody habitats. Moreover, evolution of fleshy fruits was correlated with a change to more closed (darker) habitats. An independent contrast study within Rhamnus s.l. documented allometric relations between plant and fruit size. As a phylogenetic constraint, allometric effects must be considered weak or non-existent, though, as they did not prevail among different subclades within Rhamnus s.l. Fruit size was correlated with seed size and seed number in F. alnus. This thesis suggests that frugivore selection on fleshy fruit may be important by constraining the upper limits of fruit size, when a plant lineage is colonizing (darker) habitats where larger seed size is adaptive. Phenological correlations with fruit set, dispersal, and seed size in F. alnus, suggested that the evolution of reproductive phenology is constrained by trade-offs and partial interdependences between flowering, fruiting, dispersal, and recruitment phases. Phylogenetic constraints on the evolution of phenology were indicated by a lack of correlation between flowering time and seasonal length within Rhamnus cathartica and F. alnus, respectively. On the other hand, flowering time was correlated with seasonal length among Rhamnus s.l. species. Phenological differences between biotically and wind pollinated angiosperms also suggested adaptive change in reproductive phenology.

Recruitment limitation constrains local species richness and productivity in dry grassland

Species coexistence and local-scale species richness are limited by the availability of seeds and microsites for germination and establishment. We conducted a seed addition experiment in seminatural grassland at three sites in southern Switzerland and repeated the experiment in two successive years to evaluate various circumstances under which seed limitation and establishment success affect community functioning. A collection of 144000 seeds of 22 meadow species including grasses and forbs of local provenance was gathered, and seeds were individually sown in a density that resembled natural seed rain. The three communities were seed limited. Three years after sowing, single species varied in emergence (0–50%), survival (0–69%), and establishment rates (0–27%). One annual and 13 perennial species reached reproductive stage. Low establishment at one site and reduced growth at another site indicated stronger microsite limitation compared to the third site. Recruitment was influenced by differences in abiotic environmental conditions between sites (water availability, soil minerals) and by within-site differences in biotic interaction (competition). At the least water-limited site, sowing resulted in an increase in phytomass due to establishment of short-lived perennials in the second and third years after sowing. This increase persisted over the following two years due to establishment of longer-lived perennials. After sowing in a wetter year with higher phytomass, however, productivity did not increase, because higher intensity of competition in an early phase of establishment resulted in less vigorous plants later on. Due to the generally favorable weather conditions during this study, sowing year had a small effect on numbers of established individuals over all species. Recruitment limitation can thus constrain local-scale species richness and productivity, either by a lack of seeds or by reduced seedling growth, likely due to competition from the established vegetation.

Gastropod Seed Dispersal: An Invasive Slug Destroys Far More Seeds in Its Gut than Native Gastropods

Seed dispersal is one of the most important mechanisms shaping biodiversity, and animals are one of the key dispersal vectors. Animal seed dispersal can directly or indirectly be altered by invasive organisms through the establishment of new or the disruption of existing seed dispersal interactions. So far it is known for a few gastropod species that they ingest and defecate viable plant seeds and consequently act as seed dispersers, referred to as gastropodochory. In a multi-species experiment, consisting of five different plant species and four different gastropod species, we tested with a fully crossed design whether gastropodochory is a general mechanism across native gastropod species, and whether it is altered by the invasive alien slug species Arion lusitanicus. Specifically, we hypothesized that a) native gastropod species consume the seeds from all tested plant species in equal numbers (have no preference), b) the voracious invasive alien slug A. lusitanicus – similarly to its herbivore behaviour – consumes a higher amount of seeds than native gastropods, and that c) seed viability is equal among different gastropod species after gut passage. As expected all tested gastropod species consumed all tested plant species. Against our expectation there was a difference in the amount of consumed seeds, with the largest and native mollusk Helix pomatia consuming most seeds, followed by the invasive slug and the other gastropods. Seed damage and germination rates did not differ after gut passage through different native species, but seed damage was significantly higher after gut passage through the invasive slug A. lusitanicus, and their germination rates were significantly reduced.

Piriformospora indica and Sebacina vermifera increase growth performance at the expense of herbivore resistance in Nicotiana attenuata

A Sebacinales species was recovered from a clone library made from a pooled rhizosphere sample of Nicotiana attenuata plants from 14 native populations. Axenic cultures of the related species, Piriformospora indica and Sebacina vermifera, were used to examine their effects on plant performance. Inoculation of N. attenuata seeds with either fungus species stimulated seed germination and increased growth and stalk elongation. S. vermifera inoculated plants flowered earlier, produced more flowers and matured more seed capsules than did non-inoculated plants. Jasmonate treatment during rosette-stage growth, which slows growth and elicits herbivore resistance traits, erased differences in vegetative, but not reproductive performance resulting from S. vermifera inoculation. Total nitrogen and phosphorous contents did not differ between inoculated and control plants, suggesting that the performance benefits of fungal inoculation did not result from improvements in nutritional status. Since the expression of trypsin proteinase inhibitors (TPI), defensive proteins which confer resistance to attack from Manduca sexta larvae, incur significant growth and fitness costs for the plant, we examined the effect of S. vermifera inoculation on herbivore resistance and TPI activity. After 10 days of feeding on S. vermifera-inoculated plants, larval mass was 46% higher and TPI activity was 48% lower than that on non-inoculated plants. These results suggest that Sebacina spp. may interfere with defense signaling and allow plants to increase growth rates at the expense of herbivore resistance mediated by TPIs.

Endozoochory by slugs can increase bryophyte establishment and species richness

Herbivory can affect plant community composition and diversity by removing biomass and reducing light competition. Herbivory may particularly benefit low growing species such as bryophytes, which are frequently limited by light competition. Gastropods are important herbivores of seed plants and cryptogams, furthermore, they can disperse propagules such as seeds and spores via endozoochory. However, whether gastropod herbivory can reduce the dominance of vascular plants and thereby promote the germination and establishment of endozoochorously dispersed bryophyte spores has never been tested experimentally. Moreover, it is unclear whether these possible interacting effects can influence bryophyte species richness. Here, we tested for endozoochorous spore dispersal by slugs, in combination with sowing of vascular plants, in a fully factorial common garden experiment. Enclosures contained either slugs previously fed with bryophyte sporophytes, control slugs, or no slugs. After 21 days the bryophyte cover was on average 2.8 times higher (3.9 versus 1.4) and after eight months the bryophyte species richness 2.6 times higher (5.8 versus 2.2) in enclosures containing slugs previously fed with bryophyte sporophytes than in the other treatments. Furthermore, after eight months high vascular plant cover reduced bryophyte diversity. On average enclosures without seed sowing harboured 1.6 times more bryophyte species than the ones with seed sowing (4.2 versus 2.6), indicating competitive effects of vascular plants on bryophytes. Our findings suggest that slugs are important dispersal vectors for bryophytes and that they can increase bryophyte populations and maintain bryophyte diversity by reducing the dominance of vascular plants.

Leftovers in seed dispersal: ecological implications of partial seed consumption for oak regeneration

1. Successful seed dispersal by animals is assumed to occur when undamaged seeds arrive at a favourable microsite. Most seed removal and dispersal studies consider only two possible seed fates, predation or escape intact. Whether partial consumption of seeds has ecological implications for natural regeneration is unclear. We studied partial consumption of seeds in a rodent-dispersed oak species. 2. Fifteen percent of dispersed acorns were found partially eaten in a field experiment. Most damage affected only the basal portion of the seeds, resulting in no embryo damage. Partially eaten acorns had no differences in dispersal distance compared to intact acorns but were recovered at farther distances than completely consumed acorns. 3. Partially eaten acorns were found under shrub cover unlike intact acorns that were mostly dispersed to open microhabitats. 4. Partially eaten acorns were not found buried proportionally more often than intact acorns, leading to desiccation and exposure to biotic agents (predators, bacteria and fungi). However, partial consumption caused more rapid germination, which enables the acorns to tolerate the negative effects of exposure. 5. Re-caching and shrub cover as microhabitat of destination promote partial seed consumption. Larger acorns escaped predation more often and had higher uneaten cotyledon mass. Satiation at seed level is the most plausible explanation for partial consumption. 6. Partial consumption caused no differences in root biomass when acorns experienced only small cotyledon loss. However, root biomass was lower when acorns experienced heavy loss of tissue but, surprisingly, they produced longer roots, which allow the seeds to gain access sooner to deeper resources. 7.Synthesis. Partial consumption of acorns is an important event in the oak regeneration process, both quantitatively and qualitatively. Most acorns were damaged non-lethally, without decreasing both dispersal distances and the probability of successful establishment. Faster germination and production of longer roots allow partially eaten seeds to tolerate better the exposure disadvantages caused by the removal of the pericarp and the non-buried deposition. Consequently, partially consumed seeds can contribute significantly to natural regeneration and must be considered in future seed dispersal studies.

Cysteine-proteases and cystatins from barley: molecular and functional characterization in housekeeping and defense processes

Plant cysteine-proteases (CysProt) represent a well-characterized type of proteolytic enzymes that fulfill tightly regulated physiological functions (senescence and seed germination among others) and defense roles. This article is focused on the group of papain-proteases C1A (family C1, clan CA) and their inhibitors, phytocystatins (PhyCys). In particular, the protease–inhibitor interaction and their mutual participation in specific pathways throughout the plant's life are reviewed. C1A CysProt and PhyCys have been molecularly characterized, and comparative sequence analyses have identified consensus functional motifs. A correlation can be established between the number of identified CysProt and PhyCys in angiosperms. Thus, evolutionary forces may have determined a control role of cystatins on both endogenous and pest-exogenous proteases in these species. Tagging the proteases and inhibitors with fluorescence proteins revealed common patterns of subcellular localization in the endoplasmic reticulum–Golgi network in transiently transformed onion epidermal cells. Further in vivo interactions were demonstrated by bimolecular fluorescent complementation, suggesting their participation in the same physiological processes.