Orientador: Tatiana Maria Rodrigues / Banca: Camila Rozindo Dias Milanez / Banca: Aline Redondo Martins / Resumo Espaços secretores estão distribuídos por todo o eixo vegetativo de Copaifera langsdorffii. São constituídos por epitélio secretor unisseriado e lúmen amplo e, devido à secreção produzida, representam um mecanismo de defesa constitutivo contra herbívoros e patógenos. Nas plântulas desta espécie, cavidades secretoras de óleo ocorrem no mesofilo dos eofilos e no córtex do epicótilo e hipocótilo, enquanto que canais secretores de oleorresina estão presentes na medula do epicótilo e hipocótilo. Evidências sugerem que fatores exógenos podem influenciar na produção da secreção nessa espécie; entretanto, nada se conhece sobre a influência desses fatores no desenvolvimento de canais e cavidades. Este trabalho teve como objetivo investigar a influência da luz e da temperatura na densidade e na área do lúmen de cavidades e canais secretores em plântulas de C. langsdorffii, além de estudar a influência da temperatura na ultraestrutura das células epiteliais. Plântulas foram mantidas em câmara climática sob 2500lux, 1250lux e 250lux e 15ºC, 25ºC e 35ºC. Para análises anatômicas quantitativas, amostras do epicótilo e eofilos foram processadas segundo técnicas usuais em anatomia vegetal. Para análises ultraestruturais, amostras de eofilos foram preparadas de acordo com técnicas convencionais de microscopia eletrônica de transmissão. Plântulas mantidas sob 25ºC/1250lux mostraram maior densidade de cavidades nos eofilos e no epicótilo, enquanto aquelas mantidas sob 25ºC/2500lux apresentaram maior densidade de ... / Abstract Secretory spaces are distributed throughout the vegetative axis of Copaifera langsdorffii. They are constituted by uniseriate secretory epithelium and wide lumen, and represent a constitutive defense mechanism against herbivores and pathogens due to the produced secretion. In C. langsdorffii seedlings, oil cavities occur in the mesophyll of eophylls and in the cortex of epicotyl and hypocotyl, whereas oleoresin canals are present in the pith of epicotyl and hypocotyl. Evidences suggest that exogenous factors can influence the production of secretion in this species; however, nothing is known about these factors influencing the development of canals and cavities. This work aimed to investigate the influence of light and temperature on the density and lumen area of secretory canals and cavities in seedlings of C. langsdorffii, besides to study the influence of the temperature on the ultrastructural features of epithelial cells. Seedlings were maintained in climatic chamber under 2500lux, 1250lux and 250lux, and 15ºC, 25ºC and 35ºC. For quantitative anatomical analysis, samples of epicotyl and eophylls were processed according to usual techniques in plant anatomy. For ultrastructural studies, samples of eophylls were prepared following conventional techniques of transmission electron microscopy. Seedlings under 25ºC/1250lux presented higher density of cavities in eophylls and epicotyl, whereas seedlings under 25ºC/2500lux showed higher density of canals. This differential answer to light can be related to the distinctive origin of these secretory structures. Concerning the lumen area, variable answers were observed to cavities and canals. Ultrastructurally, epithelial cells of seedlings under 25ºC showed evidences of intense secretory activity. Alterations in mitochondria, plastids and endoplasmic reticulum were observed in seedlings under 15ºC and 35ºC. In seedlings under 35ºC, the epithelial cells showed evident signals of lysis, ... / Mestre

During the summer of 1974 chemical conditions in the lower reaches of the Willamette River, Oregon were similar to those in other rivers currently experiencing nuisance algal growth problems. Temperature and chemical nutrients are not limiting. Relatively high populations of phytoplankton and productivity values for upstream periphyton beds and surface waters suggest moderately eutrophic conditions. However, with increased depth in the lower river, and a constant euphotic zone, the amount of photosynthetically available light is reduced. With sufficient depth and complete mixing the critical depth is exceeded. Primary productivity rates are subsequently limited by low light availability in the lower river.

No description available.

Environmental stress conditions can drastically affect plant growth and productivity. In contrast to soil moisture or salinity that can gradually change over a period of days or weeks, changes in light intensity or temperature can occur very rapidly, sometimes over the course of minutes or seconds. So, in our study we have taken an metabolomics approach to identify the rapid response of plants to light stress. In the first part we have focused on the ultrafast (0-90 sec) metabolic response of local tissues to light stress and in the second part we analyzed the metabolic response associated with rapid systemic signaling (0-12 min). Analysis of the rapid response of Arabidopsis to light stress has revealed 111 metabolites that significantly alter in their level during the first 90 sec of light stress exposure. We further show that the levels of free and total glutathione accumulate rapidly during light stress in Arabidopsis and that the accumulation of total glutathione during light stress is dependent on an increase in nitric oxide (NO) levels. We further suggest that the increase in precursors for glutathione biosynthesis could be linked to alterations in photorespiration, and that phosphoenolpyruvate could represent a major energy and carbon source for rapid metabolic responses. Taken together, our analysis could be used as an initial road map for the identification of different pathways that could be used to augment the rapid response of plants to abiotic stress. In addition, it highlights the important role of glutathione in initial stage of light stress response. Light-induced rapid systemic signaling and systemic acquired acclimation (SAA) are thought to play an important role in the response of plants to different abiotic stresses. Although molecular and metabolic responses to light stress have been extensively studied in local leaves, and to a lesser degree in systemic leaves, very little is known about the metabolic responses that occur in the different tissues that connect the local to the systemic leaves. These could be important in defining the specificity of the systemic response as well as in supporting the propagation of different systemic signals, such as the reactive oxygen species (ROS) wave. Here we report that local application of light stress to one rosette leaf resulted in a metabolic response that encompassed local, systemic and transport tissues (tissues that connect the local and systemic tissues), demonstrating a high degree of physical and metabolic continuity between different tissues throughout the plant. We further show that the response of many of the systemically altered metabolites could be associated with the function of the ROS wave, and that the level of eight different metabolites is altered in a similar way in all tissues tested (local, systemic, and transport tissues). These compounds could define a core metabolic signature for light stress that propagates from the local to the systemic leaves. Taken together, our findings suggest that metabolic changes occurring in cells that connect the local and systemic tissues could play an important role in mediating rapid systemic signaling and systemic acquired acclimation to light stress.

The position of newly expanded shoot leaves in the canopy of apple trees had significant and similar effects on specific leaf weight (SLW), leaf thickness, length of palisade tissue, and number of cell layers in the palisade tissue. Inside canopy leaves had lower SLW, leaf thickness, palisade length, and number of palisade cell layers than middle and outside leaves at each of six sample dates ranging from late May to early October. Outside canopy leaves had the highest values at all sample dates. Differences in SLW, leaf thickness, and palisade length between inside and outside leaves increased as the season progressed, primarily due to a general increasing trend in outside leaves that developed later in the season. Effects of canopy position on spur leaves were not as great as with shoot leaves. Regression analysis showed SLW to be significantly correlated with leaf thickness and palisade length. / M.S.

This study examined floral initiation in Rudbeckia hirta at the biochemical, cellular, and whole plant levels. Histological and histochemical examination of floral initiation revealed that the pattern of initiation followed closely that described in other species. The primary difference was in the length of time over which initiation and differentiation occurred. When subjected to limited inductive photoperiods, R. hirta responded with a delay in flowering if the plants were returned to short days (SD) before bract initiation. Increased exposure to long days (LD) increased stem height and enhanced floral development. A limited induction period of at least 8 LD allowed enough of the floral stimulus to be translocated to the meristem to cause no interruption in development even upon return to non-inductive conditions. An inhibition of development occurred only when plants were returned to SD before periclinal divisions in the pith rib meristem commenced after approximately 8 LD. Axillary bud development and final plant height were dependent on the number of inductive LD received. Polyamines have been linked to floral initiation and, in this study, were strongly correlated to the stage of floral initiation. As initiation progressed, the observed increases in putrescine and spermidine were followed by a decrease after 16 LD, the observed onset of floral development. This was contrary to that previously observed in SD plants but followed a pattern similar to that reported for cytokinin behavior. Exogenous cytokinins have been used to stimulate floral initiation in several species but Rudbeckia hirta did not respond to benzyladenine (BA) applied at the onset of LD. Floral initiation has been found to begin after six to eight LD and, in most species, BA was most effective when applied during initiation. In an attempt to increase uptake, BA was dissolved in dimethyl sulfoxide (DMSO). This did not enhance the effects of BA and, in fact, DMSO was found to be toxic at concentrations of 25% or more. / Ph. D.

Pure cultures of Saprolegnia ferax (Gruith) Thuret, Achlya americana Humphrey, and two isolates of Saprolegnia parasitica Coker were grown in a chemically defined medium under controlled conditions of temperature and illumination. Light inhibited growth and oogonium formation rates in S. ferax and A. americana. Light inhibited zoospore germination rates in A. americana and the growth rate of S. parasitica isolate 2-27-59; S. parasitica isolate 6-28-60 was indifferent to light. Neither isolate of S. parasitica formed sexual structures during the experiments. Differences in light reactions and growth rates of these two isolates indicated physiological strain differences. Photoreactivation was demonstrated in S. ferax and both isolates of S. parasitica. Evidence was shown that light caused production of toxic substances in the medium which caused inhibition of growth and sporulation. The blue peroxychromic test for hydrogen peroxide in the medium was negative but apparent catalase activity was demonstrated. Using a simple medium consisting of glucose, soluble starch and yeast extract, white light stimulated vegetative growth in S. ferax and A. americana. A quantitative distribution pattern of oogonium formation was shown. / Ph. D.

Responses of day-neutral (DN) strawberries (Fragaria X ananassa Duch.) to photoperiod and temperature under controlled environment conditions were studied and compared to those of Junebearers (JB) and everbearers (EB). At a constant 21°C, DN cultivars flowered under 9 hour days (SD), night interrupted SD (NI) and 16 hour days (LD). Runner production occurred under NI and LD but not SD. Net photosynthesis (Pn) expressed on a leaf area basis, tended to be higher for DN than JB or EB. Pn on a whole plant basis, was greater under NI than SD due to increased leaf area under NI. Pn under LD was not considered. Under fluctuating temperatures (18°/14°, 22°/18°, 26°/22° or 30°/26° C day/night combinations), responses to SD and NI varied. At 18/14, DN cultivars flowered under either NI or SD. At 22/18 and 26/22, flowering was greater under NI than SD. At 30/26, flowering did not occur. At 18/14, 22/18, or 30/26, runner production was unaffected by photoperiod and occurred under NI and SD. At 26/22, runner production was enhanced by NI compared to SD. Data for the Junebearers and everbearers is also presented and discussed. / Master of Science

Thesis (MSc (Wine Biotechnology))--University of Stellenbosch, 2006. / Plants have evolved photosynthetic systems to efficiently harvest sunlight energy for the production of carbohydrates, but these systems also are extremely susceptible to an excess of light. To combat the potential damaging effects of light, plants have developed various mechanisms to control and cope with light stress. These mechanisms include the movement of either leaves, cells (negative phototaxis) or chloroplasts to adjust the light-capturing potential, the adjustment of the light-harvesting antenna size through gene expression or protein degradation, the removal of excess excitation energy either through an alternative electron transport pathway or as heat. However, the latter mechanism based on thermal dissipation, remains the most effective to rid the plant of damaging excess light energy. This process involves several carotenoid pathway pigments, specifically the de-epoxidised xanthophyll cycle pigments. The process and extent of thermal dissipation in plants can be measured and quantified as non-photochemical quenching (NPQ) of chlorophyll fluorescence by using well-established methodologies. Several Arabidopsis and Chlamydomonas mutants affected in the xanthophyll cycle have been isolated. These mutants have provided evidence for the correlation between the de-epoxidised xanthophyll cycle pigments and NPQ as well as better understanding of the operation of the xanthophyll cycle and the related carotenoid biosynthetic enzymes. This key photoprotective role of the xanthophyll cycle is therefore a promising target for genetic engineering to enhance environmental stress tolerance in plants. Several genes from the carotenoid biosynthetic pathway of grapevine (Vitis vinifera L.) were isolated previously in our laboratory. The main aim of this study was to over-express two xanthophyll cycle genes from grapevine in Arabidopsis and to analyse the transgenic population with regards to pigment content and levels as well as certain photosynthetic parameters. The transgenic lines were compared with wild type Arabidopsis (untransformed) plants and two xanthophyll cycle mutants under non-limiting conditions as well as a stress condition, specifically a high light treatment to induce possible photodamage and photoinhibition. Transgenic Arabidopsis lines over-expressing the two V. vinifera xanthophyll cycle genes, β-carotene hydroxylase (VvBCH) and zeaxanthin epoxidase (VvZEP), were established following Agrobacterium transformation. In addition to the untransformed wild type, two NPQ mutants, npq1 (lacking violaxanthin de-epoxidase) and npq2 (lacking zeaxanthin epoxidase), were used as controls throughout this study. The transgenic lines were propagated to a homozygous T3-generation, where stable integration and expression of the transgenes were confirmed in only 16% and 12% for VvBCH and VvZEP lines, respectively. No phenotypical differences could be observed for the transgenic lines compared to the wild type, but the npq2 mutant showed a stunted and ‘wilty’ phenotype, as was previously described. To evaluate the pigment composition of the transgenic lines a reliable and reproducible method was needed to analyse carotenoids from leafy material. To this end a new high-performance liquid chromatography (HPLC) method was developed for the quantitative profiling of eight major carotenoids and chlorophyll a and b. Emphasis was placed on baseline separation of the xanthophyll pigments, lutein and zeaxanthin as well as the cis- and trans-forms of violaxanthin and neoxanthin. The method effectively distinguished Arabidopsis wild type plantlets from the two NPQ mutant lines (npq1 and 2) and could possibly find application for green leafy tissue samples in general. The carotenoid content of the NPQ mutants were in accordance with previous reports. The lack of zeaxanthin epoxidase activity in the npq2 mutant resulted in the accumulation of zeaxanthin under both low and high light conditions. This high level zeaxanthin was found to cause an initial rapid induction of NPQ at low to moderate light intensities, but this difference disappeared at high light, where zeaxanthin formation induced considerable NPQ in the wild type. Similarly, the npq1 mutant was unable to de-epoxidise violaxanthin to zeaxanthin under high light conditions, which resulted in severe inhibition of NPQ induction. Furthermore, these mutant plantlets were shown to be more susceptible to photoinhibition compared to that of the wild type. The over-expression of VvBCH resulted in a marked increase in the xanthophyll cycle pool pigments (violaxanthin, antheraxanthin and zeaxanthin) and reduced β-carotene levels under both low and high light conditions compared to that the wild type, indicating elevated β-carotene hydroxylase activity possibly due to over-expression of the VvBCH gene. Similar to the induction of NPQ in the npq2 mutant, the increased levels of zeaxanthin in the VvBCH lines did not offer any additional photoprotection. This would suggest that the heightened zeaxanthin levels observed for the VvBCH lines do not necessarily enhance photoprotection, however may protect the thylakoid membrane against lipid peroxidation as has been shown previously. The VvZEP lines however, showed reduce levels of zeaxanthin in high light conditions to that of the wild type, probably due to the competing epoxidation and de-epoxidation reactions of the xanthophyll cycle. This reduction in zeaxanthin synthesis in the VvZEP lines resulted in significant reduced NPQ induction compared that of the wild type, a phenomenon also observed for the npq1 mutant. Similar to the npq1 mutant, these lines displayed significantly increased photoinhibition, which may be due to photodamage of the reaction centers if one considers the lowered photosystem II photochemistry efficiency and reaction center openness of these lines compared to the wild type. This may suggest that even small reductions in zeaxanthin amounts can result in an increase in photoinhibition, under high light conditions. This study and its results provide fundamental information regarding two grapevine-derived carotenoid pathway genes and their possible physiological roles. Moreover, studies like these provide information that is essential when possible biotechnological approaches are planned with this central plant metabolic pathway in mind. The results highlighted the complex regulation of this pathway, necessitating attention to flux control, simultaneous manipulation of several pathway genes, and the measurement of other compounds derived from this pathway when evaluating the possible applications of the carotenoid pathway of plants.

A large number of weed seeds in the soil persist because of seed dormancy, and depletion of the seed bank through manipulation of seed dormancy has been suggested as one of the goals in weed control. This study was designed to investigate some of the factors which control dormancy and germination in Amaranthus retroflexus seeds. Germination studies were conducted at different temperatures, and either in continuous white light or in the dark. Higher temperatures increased germination and, although light interacted with temperature, its effect on germination varied with the temperature. In an attempt to determine changes in dormancy during dry storage, two lots of seeds were stored dry at different temperatures. Loss in dormancy increased with an increase in storage temperature and duration, but the time required for maximum germination varied according to the seedlot. Seeds germinated to higher percentages at high temperatures, but storage at higher temperatures and for prolonged duration resulted in seeds gaining the ability to germinate at lower temperatures. Changes in dormancy under field conditions were also examined. Seeds were buried at different depths and for different durations and they all lost viability with time, but this loss was greater in surface-sown and shallowly buried seeds. Dormancy was broken during cold periods and induced as warmer periods progressed. The effects of chemical stimulants on dormancy and germination were investigated. The response of seeds to ethephon and nitrate were assessed at different temperatures either at continuous white light or in the dark. Germination increased with the concentration of the chemicals, and a greater response was observed at lower temperatures. The response to light varied depending on temperature / Master of Science (Hons)