Effect of flooding on survival and initial growth of Ocotea pulchella (Nees) Mez. seedlings in semi-controlled light conditions

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Avaliação do potencial de cultivo in vitro de Halopteris scoparia (Linnaeus), Sauvageau (1903), Heterokontophyta (Phaeophyceae)

Dissertação de Mestrado em Biodiversidade e Biotecnologia Vegetal.

Dynamic visual information plays a critical role for spatial navigation in water but not on solid ground.

In the Morris water maze (MWM) task, proprioceptive information is likely to have a poor accuracy due to movement inertia. Hence, in this condition, dynamic visual information providing information on linear and angular acceleration would play a critical role in spatial navigation. To investigate this assumption we compared rat's spatial performance in the MWM and in the homing hole board (HB) tasks using a 1.5 Hz stroboscopic illumination. In the MWM, rats trained in the stroboscopic condition needed more time than those trained in a continuous light condition to reach the hidden platform. They expressed also little accuracy during the probe trial. In the HB task, in contrast, place learning remained unaffected by the stroboscopic light condition. The deficit in the MWM was thus complete, affecting both escape latency and discrimination of the reinforced area, and was thus task specific. This dissociation confirms that dynamic visual information is crucial to spatial navigation in the MWM whereas spatial navigation on solid ground is mediated by a multisensory integration, and thus less dependent on visual information.

Photosensitizing effects of m-tetrahydroxyphenylchlorin on human tumor xenografts: correlation with sensitizer uptake, tumor doubling time and tumor histology.

The photodynamic effects of m-tetrahydroxyphenylchlorin (mTHPC) were assessed on human malignant mesothelioma, squamous cell carcinoma and adenocarcinoma xenografts grown in nude mice and were correlated with mTHPC uptake, histology and doubling time of the tumors. Non-thermal laser light was delivered to the tumor as surface radiation 4 days after intraperitoneal administration of 0.1 and 0.3 mg mTHPC/kg body weight, respectively. The extent of tumor necrosis was measured by histomorphometry. The mTHPC concentration in non-irradiated tumors was assessed by high-performance liquid chromatography (HPLC). The tumors were graded according to their doubling time and their vascular architecture as assessed by histology. The 0.1 mg/kg dose of mTHPC resulted in an equal uptake for all 3 tumor types but revealed a larger extent of photosensitized necrosis for adenocarcinoma, which displayed a delicate tumor stroma with numerous small capillary vessels, than for mesothelioma and squamous cell carcinoma, which were both poor in stroma and vessels. The 0.3 mg/kg dose of mTHPC resulted in a 2-fold higher tumor uptake for all 3 tumor types and in a larger extent of necrosis for mesothelioma and squamous cell carcinoma, but not for adenocarcinoma xenografts, compared with the lower drug dose. Our results demonstrate that different tumor xenografts respond differently to mTHPC-PDT for a given drug-light condition. In this setting, the photosensitizing effect was more closely related to the vascular architecture of the tumors than to the sensitizer uptake and doubling time of the different tumors

Regulation of photosynthetic electron transfer in plant chloroplasts

This thesis focuses on the molecular mechanisms regulating the photosynthetic electron transfer reactions upon changes in light intensity. To investigate these mechanisms, I used mutants of the model plant Arabidopsis thaliana impaired in various aspects of regulation of the photosynthetic light reactions. These included mutants of photosystem II (PSII) and light harvesting complex II (LHCII) phosphorylation (stn7 and stn8), mutants of energy-dependent non-photochemical quenching (NPQ) (npq1 and npq4) and of regulation of photosynthetic electron transfer (pgr5). All of these processes have been extensively investigated during the past decades, mainly on plants growing under steady-state conditions, and therefore many aspects of acclimation processes may have been neglected. In this study, plants were grown under fluctuating light, i.e. the alternation of low and high intensities of light, in order to maximally challenge the photosynthetic regulatory mechanisms. In pgr5 and stn7 mutants, the growth in fluctuating light condition mainly damaged PSI while PSII was rather unaffected. It is shown that the PGR5 protein regulates the linear electron transfer: it is essential for the induction of transthylakoid ΔpH that, in turn, activates energy-dependent NPQ and downregulates the activity of cytochrome b6f. This regulation was shown to be essential for the photoprotection of PSI under fluctuations in light intensity. The stn7 mutants were able to acclimate under constant growth light conditions by modulating the PSII/PSI ratio, while under fluctuating growth light they failed in implementing this acclimation strategy. LHCII phosphorylation ensures the balance of the excitation energy distribution between PSII and PSI by increasing the probability for excitons to be trapped by PSI. LHCII can be phosphorylated over all of the thylakoid membrane (grana cores as well as stroma lamellae) and when phosphorylated it constitutes a common antenna for PSII and PSI. Moreover, LHCII was shown to work as a functional bridge that allows the energy transfer between PSII units in grana cores and between PSII and PSI centers in grana margins. Consequently, PSI can function as a quencher of excitation energy. Eventually, the LHCII phosphorylation, NPQ and the photosynthetic control of linear electron transfer via cytochrome b6f work in concert to maintain the redox poise of the electron transfer chain. This is a prerequisite for successful plant growth upon changing natural light conditions, both in short- and long-term.

Effet de l'histoire lumineuse sur la sensibilité rétinienne et circadienne à la lumière

L’objectif de ce projet de recherche était de vérifier la présence de changements de sensibilité de la rétine et du système circadien suite à deux semaines d'exposition à un milieu faiblement ou fortement éclairé, dans des conditions contrôlées en laboratoire. De plus, comme un changement de sensibilité peut modifier l'ajustement du système circadien au cycle jour-nuit extérieur, nous voulions également vérifier si la phase circadienne serait modifiée par le traitement et si la vigilance et l’humeur seraient affectées. Dix sujets ont été exposés à de la lumière tamisée (70 lux [LT]) et 10 ont été exposés à de la lumière vive (3000 lux [LV]) pendant 12 jours consécutifs en laboratoire de 8h45 à 19h00 tous les jours. L’exposition à la lumière a été mesurée 5 jours avant l’entrée au laboratoire dans l’habitat naturel du sujet et pendant la période en laboratoire à l’aide de l’Actiwatch-L®. La sensibilité rétinienne a été mesurée avant et après le traitement lumineux, par un électrorétinogramme (ERG) et la sensibilité circadienne, par le test de suppression de mélatonine salivaire. Tout au long du protocole, la vigilance, la somnolence et l'humeur ont été évaluées à plusieurs moments de la journée à intervalles prédéterminés. Après 12 jours d’exposition en lumière contrôlée, l’amplitude de l’onde-a au Vmax à l’ERG photopique a diminué en LV alors qu’elle a augmenté en LT. À l’ERG scotopique, une différence de sensibilité rétinienne (log K) entre les groupes avant le traitement expérimental s’est amenuisée à la fin du traitement (p=.053). La suppression de mélatonine après 90 minutes d’exposition au test de suppression a diminué en LV alors qu’il n’y a pas eu de modification en LT, cependant cette interaction n’était pas significative (p=.16). La phase circadienne des sujets exposés à LV a été devancée de 58 minutes (p=.04) alors qu’elle a été retardée de 26 minutes en LT (p=.32). Les mesures de vigilance subjective (EVA) ont indiqué que les sujets LV se considéraient plus éveillés que les sujets LT après le traitement (p=.02). Par contre, aucune différence n’est apparue quant aux mesures de performance psychomotrice ni de l’humeur. L’histoire lumineuse n’a pas modifié la sensibilité rétinienne dans le sens prévu par les hypothèses alors qu’il y a eu une tendance vers une augmentation de la sensibilité circadienne en condition de lumière tamisée. L’amélioration de la vigilance subjective après l’exposition en LV n’a pas été soutenue par les résultats de la performance psychomotrice. L’histoire lumineuse n’a eu aucun effet sur l’humeur des sujets. Cette étude souligne l’importance d’utiliser des mesures permettant de départager les effets immédiats d’un traitement lumineux des effets à long terme autant sur le plan rétinien que circadien. Il reste également complexe d’étudier en laboratoire des changements adaptatifs qui se produisent dans le milieu naturel en raison du confinement et des modifications physiologiques et psychologiques pouvant y être associées.

Molecular characterization of nitrate reductase gene and its expression in the marine red alga Gracilaria tenuistipitata (Rhodophyta)

The enzyme nitrate reductase (NR) responsible for the conversion of nitrate to nitrite is considered to be the rate-limiting step in nitrogen assimilation. The economically important marine macroalga Gracilaria tenuistipitata presents a circadian oscillation in NR protein content and activity. In order to identify if the regulation of NR in G. tenuistipitata happens at transcriptional levels, the NR cDNA and gene were sequenced and the NR mRNA expression was studied. Analysis of the sequenced gene revealed absence of introns which is unusual for NR genes. The transcriptional profiling revealed a circadian rhythm for NR; furthermore, a rhythm was observed in constant light condition, suggesting a possible regulation by the biological clock at the mRNA levels for NR in G. tenuistipitata.

Efeito da intensidade luminosa no desenvolvimento larval do camarão-da-amazônia, Macrobrachium amazonicum

The effects of four light intensities (0; 2.8 +/- 0.9; 5,5 +/- 1,8 e 7,8 +/- 2,5 mu mol s(-1) m(-2), about 136.5 +/- 87.5; 273 +/- 43.8 e 390 +/- 125 lux, respectively) on survival, productivity, weight gain and larval development of Macrobrachium amazonicum were investigated. Four treatments with three replicate tanks were evaluated. Newly hatched larvae were held in black tanks (80.2 +/- 0.6 larvae L(-1)) filled with 50-L-brackish water (salinity of 10), in a recirculating system. Tanks were covered with shadow cloth allowing 35% and 70% light, respectively, to reach light intensities of 2.8 +/- 0.9 and 5.5 +/- 1.8 mu mol s(-1) m(-2) at the water surface. Complete absence of light (0 mu mol s(-1) m(-2)) was obtained covering the tanks with opaque black plastic, and full-light condition used no covering (7.8 +/- 2.5 mu mol s(-1) m(-2)). Observations showed that the survival rate was not affected by light intensity. Productivity and weight gain were higher under 7.8 +/- 2.5 mu mol s(-1) m(-2) light intensity than under 0 and 2.8 +/- 0.9 1 mu mol s(-1) m(-2) intensities (P<0.05). The larval development index was similar among the treatments under the different light intensities. However, from stage VII this index was increased slightly in the treatment under 7.8 +/- 2.5 mu mol s(-1) m(-2) light intensity. In conclusion, light intensity affects larval development of M. amazonicum. Values as high as 7.8 mu mol s(-1) m(-2) (about 390 lux) improve the larval performance by enhancing development, productivity and weight gain compared to lower values.

Degradação luminosa e retenção foliar dos corantes azul brilhante FDC-1 e amarelo tartrasina FDC-5 utilizados como traçadores em pulverizações

Três estudos foram conduzidos no Núcleo de Pesquisas Avançadas em Matologia (NUPAM) pertencente à UNESP/FCA - campus de Botucatu-SP, com o objetivo de avaliar a estabilidade dos corantes Azul Brilhante FDC-1 e Amarelo Tartrasina FDC-5 quanto a diferentes períodos de exposição à luz solar e contato com folhas de Eichhornia crassipes. No primeiro estudo, soluções de 0,3125, 0,625, 1,25, 2,5, 5, 10 e 20 ppm dos corantes Azul Brilhante FDC-1 e Amarelo Tartrasina FDC-5 foram acondicionadas em tubos de quartzo hermeticamente fechados e submetidos a 0, 0,5, 1, 2, 4, 6 e 10 horas de exposição à luz solar e ao escuro. Ao final de cada período, amostras de 10 mL foram retiradas dos tubos e analisadas. No segundo estudo, os tratamentos foram dispostos no esquema fatorial 2x7: duas condições luminosas (escuro e pleno sol) e sete períodos de exposição (0, 0,5, 1, 2, 4, 6 e 10 horas), com seis repetições. Com o auxílio de micropipeta, oito gotas de 5 µL das soluções Azul Brilhante e Amarelo Tartrasina a 4.000 ppm foram depositadas em placas de Petri de vidro. Após o término dos períodos de exposição, as placas foram lavadas com 50 mL de água destilada, com o objetivo de extrair o corante depositado sobre elas. No terceiro estudo, adotaram-se os mesmos tratamentos do segundo experimento, com quatro repetições, porém as soluções foram depositadas sobre as folhas de plantas de Eichhornia crassipes. Foram adotados também os mesmos procedimentos de extração dos corantes após o término dos períodos de exposição. As soluções finais obtidas nos três estudos foram submetidas à leitura óptica de absorbância em espectrofotômetro UV-visível nos comprimentos de onda de 630 e 427 nm, para os corantes Azul Brilhante FDC-1 e Amarelo Tartrasina FDC-5, respectivamente. As várias concentrações das soluções de ambos os corantes não sofreram degradação pela luz solar quando submetidas aos vários períodos de incidência luminosa nos tubos de quartzo (ambiente fechado), visto que as curvas de recuperação apresentaram equações semelhantes àquelas concentrações que foram mantidas no escuro. A mesma estabilidade também foi observada quando os corantes foram submetidos à luz solar em ambiente aberto, ou seja, nas placas de Petri. O corante Amarelo Tartrasina também se apresentou muito estável quando depositado sobre as folhas de E. crassipes, independentemente da exposição ou não à luz solar. Para o corante Azul Brilhante, ocorreram significativas perdas de 7,8 e 18,6% quando esteve depositado na superfície da folha de aguapé pelo período de 10 horas sob condições de escuro e plena luz solar, respectivamente.

Seed germination and seedling emergence of the invasive exotic species, Clausena excavata

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Escarificação ácida, temperatura e luz no processo germinativo de sementes de Senna alata (L.) Roxb

O objetivo deste trabalho foi determinar o tempo ótimo de escarificação das sementes de Senna alata em ácido sulfúrico e verificar o efeito da temperatura, em condição de luz e escuro, na germinação de sementes dessa espécie. As sementes foram imersas em ácido sulfúrico concentrado por períodos de 0, 15, 30 e 60 minutos, e o teste de germinação realizado em BOD a 25ºC, utilizando quatro repetições de 50 sementes em cada período de tempo, sendo o delineamento estatístico inteiramente casualizado com 4 tratamentos. No segundo experimento, para verificar o efeito da temperatura e da condição de luz mais adequada a germinação, utilizou-se temperaturas de 10 a 45ºC, com intervalos de 5ºC, em condição de luz fluorescente branca ou escuro contínuo (gerbox preto). Utilizou-se o delineamento experimental inteiramente casualizado, disposto em esquema fatorial 8x2, com 4 repetições, de 50 sementes cada. em todos os testes as avaliações da porcentagem e índice de velocidade de germinação foram feitas diariamente, durante 10 dias, onde as sementes foram consideradas germinadas quando apresentaram 2 mm de raiz. Os dados foram submetidos à análise de variância e as médias comparadas pelo teste de Tukey a 5% de probabilidade. A escarificação do tegumento com ácido sulfúrico durante 60 minutos, foi ideal para as sementes de Senna alata, por proporcionar maiores valores na porcentagem e velocidade de germinação. A germinação ocorre entre 15 e 40ºC, sendo consideradas fotoblásticas neutras entre 20 e 40ºC e fotoblásticas negativas preferenciais a 15ºC. O melhor desempenho germinativo foi obtido nas temperaturas de 25, 30 e 35ºC, onde ocorreram maior porcentagem e velocidade de germinação.

Growth Regulators and KNO3 on Seed Germination of Angelonia salicariifolia

Angelonia salicariifolia is an herbaceous perennial native to Brazil with ornamental potential as garden plant, cut-flower and potted plant. It has blue flowers 1.0 to 1.4 cm long, in 10-30 cm long terminal racemes. In previous studies seeds of A. salicariifolia showed a positive photoblastic behavior under constant temperatures of 10, 15, 20, 25, 30 and 35 degrees C. The present study evaluated the effects of growth regulators (100, 200, 300, 400, 500 mg L-1 of gibberellic acid and 2.25, 11.3, 22.5 mg L-1 of 6-benzylamino-purine) and potassium nitrate (0.2 and 1.0 %) on promoting its seed germination. The experiment was conducted in a completely randomized design with six replications of 25 seeds, for each treatment. Seeds from dehiscent capsules were sown on one layer of filter paper and moistened with growth regulators or KNO3 solutions. Germination was carried out at 25 degrees C +/- 1 degrees C, under continuous light or darkness. Germination (protusion of the radicle) was observed daily for 20 days. In the dark, only gibberellic acid promoted seed germination. The percentage of germination and the speed of germination index at 400 mg L-1 (47.3%; 0.86) and 500 mg L-1 (52.0%; 0.95) were significantly higher compared to 100 mg L-1 (27.8%; 0.38) and 200 mg L-1 (32.3%; 0.49). The mean germination time at 500 mg L-1 (10.0 days) was significantly smaller compared to 100 mg L-1 (11.9 days) and 200 mg L-1 (11.5 days). Under light, treatments did not differ among each other or from the control, except for 22.5 mg L-1 of 6-benzylamino-purine and potassium nitrate (1.0%), which decreased the percentage of germination and the speed of germination index compared to control. The application of growth regulators or potassium nitrate under light condition is not necessary, since these treatments did not improve germination percentage or the speed of germination index.

Ácido 2,3,5-tri-iodobenzoico (TIBA) na micropropagação de abacaxizeiro 'IAC Gomo-de-Mel'

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Intensidades luminosas e profundidade de Semeadura na emergência e desenvolvimento de espécies de plantas daninhas

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Diversidade florística e morfoanatômica de comunidades não arbóreas de cerrado sensu stricto frente ao adensamento vegetacional na ausência de fogo

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)