Adenosine triphosphate (ATP) is well recognized for its role as the primary cellular energy currency. However, studies dating back to 1929 have reshaped our understanding of ATP as not only an energy source, but also as a signaling agent. Among the most important of these discoveries are animal purinergic receptors (P2X and P2Y receptors) that perceive extracellular ATP (eATP), primarily in the nervous system. Though eATP is an established receptor agonist in animals and applied poorly hydrolyzable ATP analogs have numerous effects on growth in plants, eATP is not widely accepted as a signal in plants where no purinoceptor has been identified. Here, enriched outside-out plasma membrane vesicles were isolated and proteins labeled with a radioactive ATP analog (8N₃ATP[α²³P]) to identify a putative purinoceptor-like receptor. We used etiolated seedlings to capture proteins from plant tissue that was actively growing and used sodium carbonate washes to separate peripheral and integral membrane proteins. With this method, we have generated lists of plasma membrane ATP binding proteins, and therefore possible eATP receptors. Ectoapyrases are phosphohydrolases thought to regulate eATP in both animals and plants. Here, we also investigated the expression and role of the candidate ectoapyrases AtAPY1 and AtAPY2 in guard cells and stomatal responses. AtAPY1 and AtAPY2 transcript and protein expression was confirmed in guard cells. Early genetic studies using an apy2 knock out with induced RNAi-silencing of APY1 suggest a role for these apyrases in stomatal regulation. In response to treatment with five hours light, the apyrase-suppressed line features wider stomatal aperture when compared to WS wild-type. / text

Salinity usually reduces plant growth in terms of height and biomass, but can increase secondary metabolite production. This frequently reported observation in guayule (Parthenium argentatum Gray, Asteraceae) was investigated for possible mechanisms.Osmotic and specific ion effects of four chloride salts (CaCl2, MgCl2, KCl, and NaCl) on leaf anatomical and plant physiological parameters were studied. One-year-old plants of guayule line AZ 2 were grown under two salt concentrations (750 ppm and 1500 ppm) for each salt type (plus a control) in sand culture (semi-hydroponic) for eight weeks under controlled greenhouse conditions in Tucson, Arizona.Growth in height decreased with increasing salt concentration. Shoot dry weight, rubber, and resin contents, however, showed no significant differences between treatments, indicating no effect from either salt concentration or salt type. There was a trend for increasing rubber content with increasing salt concentration, although not statistically significant. At the same time, net CO2 gas exchange rates decreased significantly with increasing salinity.With increasing salt concentration, guayule showed osmotic effects in terms of height, indicating a lower hydraulic conductivity. Although plants of higher salt concentrations utilized significantly less water, they had the same shoot dry weights, rubber, and resin contents. Salt-stressed plants therefore achieved higher water use efficiencies. The diurnal net CO2 gas exchange rates were significantly reduced with increasing salinity; the nocturnal net CO2 gas exchange rates showed no significant difference between the treatments.Anatomically, it was found that the stomata were raised or elevated above the epidermis, and supported by upwardly curving cells. When guayule was grown under salt treatments, the trichomes were found to include deposits of material. Trichomes might act as a detoxification repository for excess ions. Although the physiological significance of raised stomata is unknown, it is hypothesized that the unique combination of raised stomata, indumentum, and multiple layers of palisade parenchyma allows for an overall high photosynthetic capacity and performance. During stress conditions such as salinity or drought, guayule might activate an internal CO2 concentrating mechanism, i.e., bicarbonate/CO2 pump, internal CO2 recycling, or PEP carboxylation activity.

Magistro darbe tiriama tolimųjų paprastosios pušies populiacijų spyglių žiotelių skaičiaus geografinio kintamumo dėsningumai. Darbo objektas – Prokazin serijos paprastosios pušies tolimųjų kilmių bandomieji želdiniai Kazlų Rūdos MMU. Darbo metodai – ištirtos 34 populiacijos, kilusios iš rytinės pušies arealo dalies. Kiekvienoje populiacijoje atsitiktinai atrinkta po 5 medžius – iš viso 170 medžių. Medžiai įvertinti pagal pasirinktus rodiklius (skersmuo, šakų kampas, šakų skaičius menturyje ir žievės forma). Nuo kiekvieno medžio paimti ūgliai analizei, po 6 spyglius nuo kiekvieno medžio (viso 1020 spygliai). Spygliai buvo analizuoti mikroskopu pagal parinktus tokius morfologinius rodiklius: spyglio ilgis, žiotelių skaičius lygioje spyglio pusėje, žiotelių eilių skaičius lygioje spyglio pusėje. Darbo rezultatai. Atlikus tyrimus Prokazin serijos paprastosios pušies tolimųjų kilmių bandomuosiuose želdiniuose ir apdorojus duomenis statistinėmis programomis gauti tokie rezultatai: žiotelių vidutinis skaičius 2 mm spyglio segmente yra 195 žiotelės; žiotelių skaičius ir spyglių ilgis priklauso nuo medžių kilmės: populiacijos kilusios iš Europos šiaurės rytų pasižymi trumpesniais spygliais ir mažesniu žiotelių skaičiumi nei kilusios iš pietų; medžių skersmuo nėra susijęs su žiotelių skaičiumi spyglio ploto vienete, tačiau yra susijęs su spyglio ilgiu ir bendru žiotelių skaičiumi spyglyje. / This Master's thesis investigated the geographical diversity patterns of Scots pine populiations on the number of needle stomata. Object of the work – Scots pine provenance trials known as the Prokazin series in Kazlu Ruda training forest enterprise. Method of the work – 34 populations of Scots pine which represent the eastern range of the species distribution, were tested. Each population represented by 5 trees, for a total of 170 trees. The trees were evaluated with regard to the following traits: diameter, branch angle, number of whorls of branches, bark in the form. Shoots were cut from the each tree and subsequently 6 needles (for a total of 1020 needles) were taken from each tree for analysis. Each needle was analyzed under a binocular microscope separately in regard to the following seven morphological traits: needle length, number of stomatal rows on the flat side of a needle, number of stomata per 2 mm of needle length on the flat side. The results of the work. The results of this study showed that the average number of stomata is 195 per 2 millimeters segment; the number of stomata and needle length in Scots pine depends on the tree of origin: the Northeast European populations have shorter needles and fewer stomata than populations from the south; tree diameter is not associated with the number of stomata per unit area, however is related to the needle length and the total number of stomata.

As drought can impinge significantly on forest health and productivity, the mechanisms by which forest trees respond to drought is of interest. The research presented herein examined the intra-specific variation in the Populus balsamifera drought response, examining the potential role of the transcriptome to configure growth, metabolism and development in response to water deficit. Amassing evidence indicates that different species of Populus have divergent mechanisms and Three lines of inquiry were pursued to investigate the intraspecific variation the drought response in P. balsamifera. First, the transcriptome responses of six genotypes of P. balsamifera were examined using Affymetrix Poplar GeneChips under well-watered and water-deficit conditions. A core species-level transcriptome response was identified. Significantly, intraspecific variation in the drought transcriptome was also identified. The data support a role for genotype-derived variation in the magnitude of P. balsamifera transcriptome remodelling playing a role in conditioning drought responsiveness. Second, the impact of drought-stress induced declines in stomatal conductance, as well as an alteration in stomatal development in two genotypes was examined. Patterns of transcript abundance of genes hypothesised to underpin stomatal development had patterns congruent with their role in modulation of stomatal development. These results suggest that stomatal development may play a role as a long-term mechanism to limit water loss from P. balsamifera leaves under conditions of drought-stress. Finally, the drought-induced metabolome of six P. balsmaifera genotypes was interrogated. Metabolite profiling reveled amino acids such as isoleucine and proline and sugars such as galactinol and raffinose were found with increased abundance, whereas TCA intermediates succinic and malic acid were found with decreased abundance in response to drought. Comparative analysis of the metabolome and the transcriptome revealed genotypic-specific variation in energy and carbohydrate metabolism. Taken together, the findings reported in this thesis form a foundation to understand the basis of intraspecific variation in the drought response in P. balsamifera. Transcripts and metabolites that contribute to within-species differences in drought tolerance were defined. These molecular components are useful targets for both future study, as well as efforts aimed at protecting and growing trees of this important species under challenging environmental conditions.

As drought can impinge significantly on forest health and productivity, the mechanisms by which forest trees respond to drought is of interest. The research presented herein examined the intra-specific variation in the Populus balsamifera drought response, examining the potential role of the transcriptome to configure growth, metabolism and development in response to water deficit. Amassing evidence indicates that different species of Populus have divergent mechanisms and Three lines of inquiry were pursued to investigate the intraspecific variation the drought response in P. balsamifera. First, the transcriptome responses of six genotypes of P. balsamifera were examined using Affymetrix Poplar GeneChips under well-watered and water-deficit conditions. A core species-level transcriptome response was identified. Significantly, intraspecific variation in the drought transcriptome was also identified. The data support a role for genotype-derived variation in the magnitude of P. balsamifera transcriptome remodelling playing a role in conditioning drought responsiveness. Second, the impact of drought-stress induced declines in stomatal conductance, as well as an alteration in stomatal development in two genotypes was examined. Patterns of transcript abundance of genes hypothesised to underpin stomatal development had patterns congruent with their role in modulation of stomatal development. These results suggest that stomatal development may play a role as a long-term mechanism to limit water loss from P. balsamifera leaves under conditions of drought-stress. Finally, the drought-induced metabolome of six P. balsmaifera genotypes was interrogated. Metabolite profiling reveled amino acids such as isoleucine and proline and sugars such as galactinol and raffinose were found with increased abundance, whereas TCA intermediates succinic and malic acid were found with decreased abundance in response to drought. Comparative analysis of the metabolome and the transcriptome revealed genotypic-specific variation in energy and carbohydrate metabolism. Taken together, the findings reported in this thesis form a foundation to understand the basis of intraspecific variation in the drought response in P. balsamifera. Transcripts and metabolites that contribute to within-species differences in drought tolerance were defined. These molecular components are useful targets for both future study, as well as efforts aimed at protecting and growing trees of this important species under challenging environmental conditions.

Diss. (sammanfattning) Umeå : Sveriges lantbruksuniversitet, 2006. / Härtill 3 uppsatser.

Thesis (Ph. D.)--Florida State University, 2005. / Advisor: William H. Outlaw Jr., Florida State University, College of Arts and Sciences, Dept. of Biological Science. Title and description from dissertation home page (viewed Jan. 26, 2006). Document formatted into pages; contains x, 60 pages. Includes bibliographical references.

Orientador: Marcos Silveira Buckeridge / Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-11T10:56:04Z (GMT). No. of bitstreams: 1 Machado_MarceloRubens_D.pdf: 11185309 bytes, checksum: d108572b634306b2f02511963c8df8c9 (MD5) Previous issue date: 2007 / Resumo: A concentração atmosférica de dióxido de carbono (CO2) aumentou de 280 ppm, à época da Revolução Industrial, para cerca de 370 ppm atualmente, e continua a aumentar cerca de 1,8 ppm ao ano. Os efeitos dessa elevação no sistema climático global têm atraído considerável atenção, mas ainda não há informações suficientes sobre como essas mudanças podem afetar as espécies arbóreas tropicais, em especial na estrutura dessas plantas, componente relevante no sucesso competitivo e fisiológico dos vegetais. Nesse estudo são apresentados dados de análises estruturais, ultra-estruturais e bioquímicas de folhas de duas espécies tropicais - o jatobá da mata Hymenaea courbaril e o do cerrado H. stigonocarpa - expostas à elevada concentração de CO2. Após cem dias de exposição em câmaras de topo aberto, as espécies demonstraram ser reativas ao aumento da [CO2], especialmente H. stigonocarpa. Essa alteração atmosférica resultou em maior crescimento das plantas, com significativa redução do número de estômatos e aumento dos parênquimas fotossinteticamente ativos, especialmente do paliçádico. A análise quantitativa da cera epicuticular indicou haver variação, porém não significativa, da cera bruta total das espécies. A cromatografia gasosa e a espectrometria de massa mostraram que a cera epicuticular das duas espécies é constituída, principalmente, por nonacosanos e triterpenódes de provável função alcoólica. Os estudos ultra-estruturais evidenciaram alterações na morfologia dos cloroplastos, em decorrência do aumento na deposição de amido, nos exemplares submetidos à elevada [CO2], mas não há evidências de que esse depósito afete o desenvolvimento do vegetal. Também houve alteração na morfologia da cera epicuticular. As técnicas microscópicas e bioquímicas adotadas mostraram-se úteis na determinação de respostas de plantas tropicais a elevadas concentrações de CO2, constituindo-se em ferramentas de aplicação viável em estudos de mudança climática / Abstract: The atmospheric concentration of carbon dioxide (CO2) increased from 280 ppm, during the Industrial Revolution period, to about 370 ppm in the nowadays. It is continuing to increase about 1.8 ppm per year. The effect of this rise in CO2, which is leading to global warming have attracted considerable attention, generating reports on how these changes can affect vegetables, in particular tropical trees species. There are few reports on the consequences of the rise in atmospheric CO2 on plant structure, even considering its relevance for the competitive and physiological success of plants. In this study we present the data about structural, ultrastructural and biochemical analyses of seedlings from two tropical species ¿ the jatobá Hymenaea courbaril and H. stigonocarpa ¿ both exposed to high CO2 concentration. After one hundred days of exposure to elevated CO2 (720 ppm) in open top chambers, the species demonstrated to be reactive, in particular the H. stigonocarpa. This atmospheric alteration resulted in slightly higher plant growth, with significant reduction of the number of stomata and increase of the photosynthetically active tissues, especially of the palissadic layer of the leaves. The quantitative analysis of the epicuticular wax indicated no variation in the species. Gas chromatography and mass spectrometry analyses have shown that the epicuticular wax of the two species is constituted mainly of nonacosans and triterpenoids of probable alcoholic function. The ultrastructural studies revealed alterations in the morphology of the epicuticular wax, and of the chloroplasts the later due to the increase in starch deposition, in the samples submitted to increased CO2 concentration. However, there were no evidences that these changes affect plant development as a whole. The microscopical and biochemical techniques used were useful in determining the responses of tropical plants to the increase of CO2 concentrations, being therefore valuable tools in studies of climatic change / Doutorado / Biologia Celular / Doutor em Biologia Celular e Estrutural

Nas últimas décadas, o aumento da demanda por produtos provenientes de plantios florestais resultou na expansão das áreas de plantio, principalmente para as regiões do Cerrado, onde as terras são mais baratas e extensas. Nessas regiões fronteiriças, as condições edafoclimáticas são diferentes das atuais regiões de plantio, apresentando regime pluviométrico irregular e escasso associado à altas temperaturas e elevada evaporação e diferentes tipos de solo, o que pode restringir o uso de genótipos de eucalipto nesses locais. Tal fato exige pesquisa acerca do efeito do estresse hídrico e da adubação potássica no comportamento morfofisiológico dos clones, uma vez que plantas bem nutridas com potássio resistem melhor ao estresse hídrico devido à maior retenção de água. Assim, o objetivo do presente trabalho foi avaliar o efeito da omissão do potássio e, conseguintemente, do déficit hídrico nas respostas fisiológicas e de crescimento de seis clones de eucalipto. Desse modo, foi instalado em dezembro de 2011 um experimento na Estação Experimental de Ciências Florestais de Itatinga (EECFI) pertencente à ESALQ/USP. O solo onde o experimento foi implantado é classificado como Latossolo Vermelho-Amarelo distrófico típico A moderado textura média. O delineamento experimental utilizado foi em blocos casualizados, com seis repetições, cada uma composta por doze tratamentos em um arranjo fatorial 6x2, sendo seis clones de eucalipto (COP 1404, COP 1407, AEC 042, AEC 224, AEC 144 e AEC 1528) e duas doses de potássio (0 kg.ha-1 e 140 kg.ha-1). As parcelas são retangulares, formadas por 25 árvores (5 linhas de 5 plantas) em espaçamento 3m x 2m. A área útil da parcela (onde efetivamente foram realizadas as avaliações) é composta pelas 9 plantas centrais. Foram realizadas avaliações das variáveis: DAP (cm), altura total (m), comprimento da copa (m), índice de área foliar (IAF) (m2/m2), área foliar específica (AFE) (m2/kg), deposição de folhedo (kg/ha), área foliar (cm2), densidade estomática (estômatos/mm2), dimensão dos estômatos (μm), volume total (m3/ha), análise nutricional do folhedo, incremento diamétrico através de cintas dendrométricas e dendrômetros de ponto eletrônicos. O efeito da omissão de potássio foi verificado para todos os clones, mas a resposta morfofisiológica dos clones foi diferente entre eles. Altura, comprimento de copa, área do limbo foliar, densidade estomática, diâmetro polar (DP) e diâmetro equatorial (DE) dos estômatos e variação cambial foram as variáveis que apresentaram efeito da interação clone-potássio. A variável DP mostrou resultados mais correlacionados com a produtividade dos clones que a variável densidade estomática, indicando que o DP influencia diretamente na eficiência de uso de água pela planta. A análise do incremento diamétrico mostrou que a atividade cambial é sazonal e que indivíduos dentro de um mesmo tratamento tem um ritmo de crescimento e responde de forma diferenciada às variações ambientais. Por fim, as variáveis morfofisiológicas que melhor explicam o maior grau de sucesso de alguns clones para resistir ao estresse hídrico são: comprimento de copa, IAF, deposição de folhedo e DP. / In the last few decades, the increased demand for products from forest plantations hás resulted in the expansion of plantation áreas, mainly for the Cerrado region, where land is cheaper and extensive. In these border regions, soil and climatic conditions are different from current planting areas, with irregular and scarce rainfall associated with high temperatures and high evaporation and different soil types, which may restrict the use of eucalyptus genotypes at these sites. This fact requires research about the effect of water stress and the potassic fertilization on the morphophysiological behavior of the clones, as well-nourished plants with potassium have stronger resistance to water stress due to the better water retention. The objective of this study was to evaluate the effect of potassium omission and consequently, of the water deficit on physiological responses and growth of six eucalyptus clones, Thus, it was installed in December 2011 an experiment in the Experimental Forest Sciences Station of Itatinga (EECFI) belonging to the ESALQ/USP. The soil where the experiment was carried out is classified as Oxisol Hapludox tipic A moderate medium texture. The experimental design was randomized blocks with six repetitions, each one consisting of twelve treatments in a factorial arrangement 6x2, with six eucalyptus clones (COP 1404, COP 1407, AEC 042, AEC 224, AEC 144 and AEC 1528) and two doses of potassium (0 kg ha-1 and 140 kg ha-1). The plots are rectangular, formed by 25 trees (5 rows of 5 plants) in 3m x 2m spacing. Each plot useful area (which were actually conducted evaluations) is composed of the 9 central plants. The variables were: DAP (cm), total height (m), canopy height (m), leaf area index (LAI) (m2/m2), specific leaf area (SLA) (m2/kg), litterfall (kg/ha), leaf area (cm2), stomatal density (stomata/mm2), size of stomata (uM), total volume (m3/ha), nutritional analysis of leaf litter, diameter increment through dendrometer bands and electronic point dendrometers. The effect of potassium omission was observed for all the clones, but the morphophysiological response were different Height, crown length, leaf area, stomatal density, stomata polar diameter (PD) and equatorial diameter (ED) and cambial variations were the variables that showed the effect of clone-potassium interaction. The variable DP showed results more correlated with the productivity of the clones than the variable stomatal density, indicating that the DP directly influences the water use efficiency of the plant.The dendrometric increment analysis showed that the cambial activity is seasonal and individuals within the same treatment has a growth rate and responds differently to environmental variations. Finally, the morphological and physiological variables that best explain the higher degree of success of some clones to resist to water stress are: length canopy, LAI, leaf litter deposition and DP.

A necessidade de expansão da eucaliptocultura em áreas com déficit hídrico, leva à necessidade de entender os processos fisiológicos relacionados ao crescimento fundamentais para possibilitar a alta produtividade das florestas nessas áreas. Este estudo teve como objetivo avaliar o efeito da adubação potássica no crescimento, respostas ecofisiológicas e nutricionais em clones de Eucalyptus grandis x Eucalyptus urophylla, para tal, instalou-se na Estação Experimental de Ciências Florestais da ESALQ/USP em Itatinga-SP, em Latossolo Vermelho-Amarelo distrófico um experimento em blocos ao acaso em arranjo fatorial 6 X 2 (seis clones e duas diferentes doses de adubação potássica) e parcelas retangulares de 25 plantas, nas quais as 9 centrais constituíram a parcela útil. A adubação de base foi uniforme em todo experimento, e aos três e oito meses de idade adubou-se as parcelas sem omissão de potássio com 50 e 78 Kg ha-1 de KCl respectivamente totalizando 140 kg ha-1 de potássio aplicados ao longo do experimento. A avaliação da absorção dos macronutrientes foi feita aos seis e quinze meses pós-plantio, em três indivíduos amostrados aleatoriamente de cada parcela. As avaliações de crescimento foram realizadas trimestralmente em todos os indivíduos da parcela, com início da medição da altura aos dois meses, área da copa aos quatro meses e DAP aos doze meses pós-plantio. A avaliação da eficiência e uso de água foi feita aos quinze meses com base na área foliar e nas características estomáticas dos clones, em quatro blocos, dos quais três árvores foram amostradas aleatoriamente. O folhedo foi coletado mensalmente em todas as parcelas, com início aos quinze meses de idade. Maiores crescimentos em altura, DAP e área da copa, foram apresentados pelos clones 2 e 6 caracterizados por apresentarem resistência ao estresse hídrico, e pelos clones 1 e 4 de moderada resistência. Sob omissão de potássio, os clones apresentam maiores densidades estomáticas acompanhadas pela redução no diâmetro dos estômatos. Houve variação do estado nutricional dos clones em função do material genético. A variação nas respostas dos clones à adubação potássica teve início aos doze meses para as variáveis de crescimento. Sob adubação potássica, os clones mostraram-se eficientes no uso da água, por apresentarem estômatos maiores em densidades menores, o que proporcionou maior crescimento. / The need for expansion of eucalyptus plantations in areas with water deficit, leads to the need to understand the physiological processes related to growth important to enable high forests productivity in these areas. In order to evaluate the effect of potassium fertilization on growth, nutritional and ecophysiological responses in Eucalyptus grandis x Eucalyptus urophylla clones an experiment was settled at the Experimental Station of Forest Sciences ESALQ/USP in Itatinga-SP in red-yellow latossols dystrophic, in a randomized block design in a factorial 2 X 6 (six clones and two different potassium fertilization doses) and rectangular plots of 25 plants, of which the 9 central plants were the useful plot. The basic fertilization was uniform throughout the experiment, and at 3 and 8 months, the plots without omission of potassium were fertilized with 50 and 78 kg ha-1 of KCl respectively, totalizing 140 kg ha-1 applied throughout the experiment. The macronutrients absorption evaluation was made at six and fifteen months after planting, in three random plants in each plot. Quarterly, growth evaluations were performed in all subjects, starting measuring heights at two months, crown area at four months and DBH at twelve months after planting. At fifteen months in four blocks, of which three trees were selected randomly, water use efficiency was evaluated, based on clone´s leaf characteristics and stomata. The litter was collected monthly in all plots, starting at 15 months. Highest increases in height, DBH and crown area were presented by clones 2 and 6 which are characterized by their resistance to water stress, and the clones 1 and 4 which are characterized by moderate resistance. Under potassium omission, the clones showed higher stomata densities accompanied by stomata diameter reduction. There was variation in the nutritional status of clones as a function of genetic material. The variation in the clone´s responses to potassium fertilization for the growth variables begin at twelve months. The clones were efficient in water use, because they have larger stomata in lower densities, which provided greater growth.