Global gene expression in Sorghum bicolor, an important crop showing drought tolerance in arid and semi-arid cultivated areas, was monitored to exposure of 8-days seedlings to water deficit (20% polyethylene glycol) or cold stress (4 ºC). A sorghum cDNA microarray, including ~13,000 (milestone version 1) or ~28,000 (milestone version 2) unigenes, was used to examine gene expression in shoots and roots at 3 and 27hours after stress treatment. ~1,300 and ~2,300 genes were modulated by water deficit and cold stress, respectively. Up-regulated genes included previously identified stressinduced genes such as early drought-induced gene, dehydrin, late embryogenesis abundant gene, glycin and proline-rich gene, and water stress-inducible genes as well as unknown genes. Genes involved in signal transduction, lipid metabolism, transporter, and carbohydrate metabolism are induced. Quantitative real-time PCR was used to quantify changes in relative mRNA abundance for 333 and 108 genes in response to water deficit and cold stress, respectively. Stress-induced genes were classified by kinetics. Eighteen of 108 cold-induced genes were modulated by cold but not by ABA and PEG treatment. This research provides the starting point for detailed analysis and comparison of water deficit and cold modulated gene networks in sorghum.

No description available.

Dry mixed conifer forests of southwestern North America are projected to be particularly vulnerable to ongoing persistent warm drought conditions, and related increases in wildfire frequency, size and severity, due in part to consequences of over a century of fire exclusion. Prescribed fire is applied actively in many landscapes to reduce hazardous fuel loads and continuity, restore forest community composition and structure, and increase tree resilience to drought stress. However, fire can also adversely affect tree growth by damaging cambial, root, and canopy tissues, leading to tradeoffs in the use of fire as a tool for forest resilience. Radial growth is an indicator of climatic and ecological stress and can thus provide a relative measure of resilience to stress and disturbances; but, the mechanisms driving tree resilience to prescribed fire and concurrent drought are poorly understood. Thinning effects of prescribed fire may increase tree resilience to drought by increasing water, light and nutrient availability and production of defense mechanisms. However, trends over the last century indicate warming temperatures are increasing tree sensitivity to fire by reducing post-fire growth (lower resilience) and increasing the likelihood of mortality. Trees can be resistant to fire exposure, and where growth changes occur they can be transient or persistent. We studied the interactions between tree- and stand-level fire effects on the growth responses of surviving Abies concolor, Pinus jefferyi, Pinus ponderosa, and Pseudotsuga menziesii over 24 years of variable climatic conditions in ten National Parks across the western and southwest United States. We used linear mixed effects models to identify mechanisms influencing resistance and resilience responses to fire and interannual climate, using climatic water deficit (CWD) as an index of climatic stress. Compared to pre-fire growth, trees exposed to fire increased growth during periods of greater water deficits. Tree growth responses were variable among and within species and size classes, but contingent on time-since-fire and the climate during the recovery period. Negative fire effects on tree resistance were generally transient, while climate and pre-existing stand conditions were persistent controls on tree resilience. These results suggest that antecedent and subsequent climate conditions modulate post-fire forest response. Consideration of climate variation could improve the strategic use of prescribed fire for tree resilience to drought, and a deeper understanding of factors contributing to prefire growth may elucidate the mechanisms driving post-fire growth responses.

No description available.

Wheat plants are affected by water-deficit stress in various regions of the world resulting in reduced crop productivity and thus decreased food production. To better understand the protein changes of water-deficit stress in wheat roots, comparative proteomics was performed using 2D gel electrophoresis followed by HPLC-MS/MS. Forty proteins were identified, twenty-nine of which are non-redundant. Cell walls root proteins were identified using SDS-PAGE followed by mass spectrometry, resulting in the identification of seventeen proteins. Cell wall polysaccharides were extracted from roots to identify the polysaccharide metabolic changes that occurred under water-deficit stress conditions. Using FT-IR, cellulose was found to increase while hemicellulose and pectin content decreased. Further analysis by PCA showed changes in overall polysaccharide content over time. Overall, proteins identified in wheat whole roots and cell walls, combined with indications of polysaccharide modifications in the root cell walls, give us a better understanding of wheat responses to water-deficit stress. / Plant Biology

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Agradecemos a compreensão. on 2017-02-24T19:33:38Z (GMT) / Submitted by Carlos Eduardo Caixeta de Castro null (carloscaixetaagro@hotmail.com) on 2017-02-24T20:41:43Z No. of bitstreams: 1 Tese 21 02 2017 .pdf: 1498537 bytes, checksum: 1cc288db2cf86a6945c74350426712f9 (MD5) / Approved for entry into archive by LUIZA DE MENEZES ROMANETTO (luizamenezes@reitoria.unesp.br) on 2017-03-06T18:54:40Z (GMT) No. of bitstreams: 1 castro_cec_dr_jabo.pdf: 1498537 bytes, checksum: 1cc288db2cf86a6945c74350426712f9 (MD5) / Made available in DSpace on 2017-03-06T18:54:40Z (GMT). No. of bitstreams: 1 castro_cec_dr_jabo.pdf: 1498537 bytes, checksum: 1cc288db2cf86a6945c74350426712f9 (MD5) Previous issue date: 2017-02-02 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Os programas de melhoramento florestal no Brasil têm o desafio de atender à demanda crescente por produtos madeireiros. Por outro lado, a ocorrência de secas sucessivas no Brasil vem criando uma instabilidade produtiva nas florestas plantadas. Com isto, este trabalho teve como objetivos: i) estimar parâmetros genéticos e avaliar a adaptabilidade, estabilidade e produtividade de famílias de polinização aberta de Eucalyptus grandis em diferentes locais no Brasil e propor estratégia de seleção para o conjunto desses locais; ii) avaliar o comportamento de uma amostra destas famílias, cultivadas em vasos em casa de vegetação, submetidas a dois regimes hídricos na presença e ausência de adubação potássica e iii) relacionar os resultados obtidos em casa de vegetação com aqueles obtidos em campo. A primeira parte do trabalho foi realizada com os dados de altura, diâmetro à altura do peito e volume de madeira, cedidos pelo Instituto de Pesquisas e Estudos Florestais (IPEF), provenientes de testes de 165 famílias de polinização aberta de Eucalyptus grandis avaliados em oito locais no Brasil. Estes dados foram submetidas à análise de adaptabilidade, estabilidade e produtividade pelo método da Média Harmônica da Performance Relativa dos Valores Genéticos (MHPRVG). Com base neste estudo, uma amostra de 15 das 165 famílias foi usada para o experimento conduzido em casa de vegetação, onde estas foram submetidas a dois regimes de irrigação, na presença e ausência de adubação potássica, avaliando-se caracteres de crescimento, nutricionais e fisiológicos. No experimento de campo, observou-se interação genótipo por ambiente, com alteração no ordenamento dos genótipos nos diferentes ambientes, conforme detectado pelo método MHPRVG comparado com as ordenações dos genótipos por local, o que indica que a seleção simultânea para todos os locais não é a melhor estratégia de melhoramento. Os locais Pratânia e Monte Dourado B podem ser utilizados para obtenção de genótipos representativos e superiores para a maior parte das regiões estudadas. No experimento em casa de vegetação, observou-se a existência de variabilidade entre as progênies de famílias de polinização aberta de E. grandis possibilitando a obtenção de sucesso na seleção de famílias superiores para as características de crescimento; em contrapartida existe pequena variabilidade nas progênies quanto às características fisiológicas e nutricionais, o que dificulta o sucesso na seleção. Porém, mesmo perante a baixa variabilidade genética, de uma forma geral, é possível obter ganhos com a seleção das famílias superiores. Ao associar os resultados para as 15 famílias avaliadas em comum, nos experimentos de campo e em casa de vegetação, verificou-se que as relações entre os dois experimentos são fracas, com grandes alterações na classificação das famílias de uma condição para outra, demonstrando a dificuldade de usar resultados de experimentos sob condições semicontroladas para predizer o comportamento em campo. / The programs of forest improvement in Brazil have the challenge of meeting this rising demand. On the other hand, the occurrence of repeated droughts in Brazil has been creating a productive instability in commercial forests. Likewise, this work had the objective: i) estimated genetic parameters and evaluating the adaptability, stability and productivity of open-pollinated families of Eucalyptus grandis in different sites in Brazil, and proposing selection strategy for the set of these sites; ii) evaluating the behavior of a sample of these families, cultivated in pots at greenhouse, subject to two hydric regimes in the presence and absence of potassium fertilizations and iii) relating the results obtained in greenhouse with those obtained in the field. The first part of this work was done with height, diameter at breast height and wood volume data, provided by the Instituto de Pesquisas e Estudos Florestais (IPEF), from tests of 165 open-pollinated families of Eucalyptus grandis evaluated in eight places. These data were submitted to analysis of adaptability, stability and productivity through the method of Harmonic Mean of the Relative Performance of the Genetic Values (HMRPGV). Based on this study, a sample of 15 out of 165 families was used for the experiments realized in greenhouse, where these were submitted to two irrigation regimes, in the presence and absence of potassium, evaluating the growth, nutritional and physiological traits. There was genotype by environment interaction with change in the order of genotypes in different environments as detected by the method HMRPFV compared with the order of genotypes in each site, which shows the simultaneous selection for all sites is not the best strategy of improvement. The sites Pratânia and Monte Dourado B can be used for obtaining representative and superior genotypes for most regions studied. Even with a small genetic variance when the selection was done, small gains were obtained. By associating the results to as 15 families in field and greenhouse experiments, it was verified that as relations between the two experiments are weak, with large changes in the classification of families from one condition to another, demonstrating a difficulty of using experiment results under semicontrolled conditions to predict field behavior.

Cyanide is produced throughout a plant's life cycle alongside the hormone ethylene by oxidation of 1-aminocyclopropane-1-carboxylic acid. Production increases during certain developmental stages such as seed germination, seedling elongation, fruit ripening and senescence. Abiotic stresses increase ethylene production giving rise to `stress cyanide'. Cyanide also comes from metabolism of cyanogenic compounds. Cyanide is however, a toxic chemical which readily binds to metallo-enzymes inhibiting primary metabolic processes. Plants have mechanisms to maintain cyanide homeostasis such as the β-cyanoalanine pathway whereby cysteine reacts with cyanide forming β--cyanoalanine, mediated by β-cyanoalanine synthase and cysteine synthase. A dual nitrilase 4 enzyme then converts the β-cyanoalanine into asparagine or aspartate and ammonium. Studies have suggested that the physiological function of the pathway is not restricted to detoxification and assimilation of excess cyanide. The overall research goal was to investigate the role of the pathway in plant tolerance to water deficit and exogenous cyanide exposure in Arabidopsis thaliana. The first objective was to investigate responsiveness of the pathway to duration and intensity of water deficit and cyanide exposure. The second was to investigate the contribution of enzymes associated with the pathway to cyanide metabolism. The questions addressed were whether there is enzymatic redundancy in enzymes associated with the first step of cyanide detoxification and whether there is pathway redundancy between the β-cyanoalanine and an alternative sulfurtransferase pathway. A. thaliana Col-0 and three SALK-line mutants with a T-DNA insertion for the genesAtCysA1, AtCysC1 and AtNIT4 were grown and exposed to water stress. Physiological and biochemical measurements were taken. The results showed a transient increase in cyanide concentration and β-cyanoalanine synthase activity on exposure to stress. The response pattern was similar regardless of intensity or duration of stress. Knocking out AtCysA1 or AtCysC1 did not impair the ability of plants to metabolize cyanide and tolerate stress i.e the enzymes were functionally redundant. The AtNIT4 mutant however, was impaired in cyanide metabolism and exhibited a sensitive phenotype under both stresses, suggesting that the cyanoalanine pathway is the sole pathway in cyanide detoxification. The results show that the pathway may be an important tool in improvement of plant tolerance to abiotic stress.

Orientadora: Sonia Marli Zingaretti / Banca: Cristina Lacerda Soares Petrarolha Silva / Banca: Janete Apparecida Desidério Sena / Resumo: A seca é um dos principais fatores abióticos que afetam a produtividade das plantas. Para detectar os genes expressos sob condições de deficiência hídrica desenvolveu-se uma análise da expressão gênica das plantas de cana-de-açúcar de uma cultivar tolerante (cv. SP83-2847) e outra sensível (cv. SP90-1638) à seca, utilizando macroarranjos de DNA para monitorar a expressão gênica de 3.575 clones de folhas de cana-de-açúcar e, a partir dos resultados obtidos foram selecionados dois ESTs (SCJFLR2036B11 e SCBFLR1005E12) que se apresentaram como diferencialmente expressos nas duas cultivares. Os clones foram seqüenciados e identificados e correspondem ao gene que codifica para a Nitrilase e para elementos genéticos móveis (Retrotransposon). Os dados de expressão gênica foram validados por meio de RT-PCR semiquantitativo e Southem blotting e a seqüência de nucleotídeos obtida foi utilizada nas buscas em bancos de dados e na comparação de seqüência o que contribuiu para a atribuição de função biológica. O gene similar ao da Nitrilase apresentou-se induzido tanto na situação de deficiência hídrica no cultivar sensível (SP90-1638) como no cultivar tolerante (SP83-2847) nas plantas não expostas ao déficit hídrico. O gene que codifica um elemento genético móvel (Retrotransposon) apresentou-se induzido no cultivar tolerante na situação controle em detrimento da situação estressada e invariável para cultivá-Io sensível nas duas situações. / Abstract: Drought is one of the main abiotic stresses which aftect plant productivity. To detect genes expressed under drought conditions, a gene expression study of sugarcane plants was performed, with drought tolerant (cv. SP83-2847) and sensitive (cv. SP90-1638) cultivars, using DNA macroarrays to monitor gene expression of 3.575 sugarcane leaf clones, and from the obtained results two ESTs (SCJFLR2036811 and SC8FLR1005E12) identified as difterentially expressed in both cultivars. were selected. The clones were sequenced and identified as the gene which codifies Nitrilase and mobile genetic elements (Retrotransposon). Gene expression data were confirmed by Semiquantitative RT-PCR and Southern blotting analysis, and the obtained nucleotide sequence used in database searches and in sequence comparisons, which contributed for the biological function attribution. The gene similar to Nitrilase appeared to be induced under water deficiency in the sensitive cultivar (SP90¬ 1638) as well as in the tolerant cultivar (SP83-2847) in plants not submitted to water deficit. The gene which codifies a mobile genetic element (Retrotransposon) appeared to be induced in the tolerant cultivar under the controlled condition in detriment of the stress, and unchanged for the sensitive cultivar under both conditions. / Mestre

The Virginia-Carolina (VC) region including Virginia, North Carolina, and South Carolina, is the most important peanut production region for the large seeded, virginia-type peanut in the United States.  In recent years, an increased frequency of heat and drought episodes with significant effects on peanut yield was observed in the VC region.  Because limited information is available on the mechanisms virginia-type peanut develops in response to heat and water stress, the present study evaluated several physiological and metabolic characteristics and their relationship with yield for eight cultivars and breeding lines.  Experiments were conducted under rainfed and irrigated field trials in 2011 and 2012, and in a growth chamber under optimum (30/25 "C) and high temperature (40/35 "C) conditions.  The long term goal of this study is to help development of more tolerant peanut cultivars to heat and drought in the VC region. Visible symptoms of water-deficit stress were observed in peanut during the field experiments in both years.  Significant (p d 0.05) variations for yield, membrane injury, chlorophyll fluorescence (Fv/Fm ratio), specific leaf area, SPAD chlorophyll content, and relative levels of polar and non-polar metabolites were observed in response to water regime, growth stage, and genotype in both years during the field studies.  Similarly each year, the Fv/Fm ratio, organic acids, and saturated fatty acids decreased in rainfed vs. irrigated plants, while the sugar and sugar alcohol relative levels increased.  Regardless the water regime, lower levels of saturated fatty acids and sugars, and higher levels of unsaturated fatty acids and sugar alcohols were associated (p < 0.05) with higher pod yield in field conditions.  Genotypes Phillips, SPT06-07, and N05006 showed potential tolerance and N04074FCT, CHAMPS, and Bailey susceptibility to water deficit in field studies.  Significant physiological and metabolic changes were also observed in response to heat stress under controlled conditions in peanut seedlings.  A general decrease in organic acid and saturated fatty acid levels and an increase in membrane injury, sugar, and unsaturated fatty acid levels were observed under both water deficit and heat stress conditions.  Overall, results from both experiments were suggestive of natural stress responses rather than adaptive mechanisms to water deficit and heat stress of the virginia-type genotypes used in this study. Among all genotypes, SPT 06-07 showed improved tolerance to both stresses.  Our results suggest that monitoring chlorophyll fluorescence and changes in the levels of selected metabolites can be used to screen new peanut lines for drought and heat stress tolerance. / Master of Science in Life Sciences

Dans le contexte des changements climatiques, la capacité d’adaptation à la contrainte hydrique des arbres de plantation devient un enjeu majeur pour le maintien de leur productivité. La plasticité phénotypique des génotypes, facteur majeur de l’adaptation aux changements environnementaux, reste encore insuffisamment décrite chez les arbres forestiers, plantes pérennes à long cycle de révolution. Cette thèse se propose i / de caractériser la plasticité phénotypique pour deux clones commerciaux d’eucalyptus soumis à une contrainte hydrique au champ, ii/ d’identifier les caractères potentiellement adaptatifs (i.e. ceux présentant de l’interaction GxE) et iii/ de mettre en évidence les mécanismes moléculaires sous-jacents. Un dispositif expérimental mis en place en république du Congo a permis de comparer, à travers une approche intégrant différents caractères allant de l'expression de gènes à la production de biomasse, la réponse de deux génotypes d’eucalyptus soumis à des régimes hydriques différents au cours de la saison sèche sur les deux 1ères années de croissance. Les résultats montrent que si l’effet de la saison sèche est relativement similaire pour les deux génotypes (réduction des accroissements relatifs), le clone plus productif présente des accroissements très élevés en saison des pluies, et semble donc mieux optimiser des conditions environnementales favorables. La capacité d’ajustement physiologique, anatomique et moléculaire au niveau foliaire de ce génotype semble être un atout au maintien de ses capacités photosynthétiques.Cette thèse a permis de mettre en évidence certains critères (surface foliaire spécifique, accumulation de phénols, épaisseur de collenchyme dans les feuilles), qui pourraient permettre d’évaluer le potentiel adaptatif des populations d’eucalyptus, nécessaire à leur gestion durable des plantations. D’autre part, certains gènes impliqués dans la photosynthèse et le métabolisme secondaire, dont l’expression pourrait être liée à la variation de caractères phénotypiques ont été mis en évidence. L’identification de ces gènes constitue une première étape vers la compréhension des mécanismes moléculaires sous-jacents à la plasticité phénotypique chez l’eucalyptus. / In the context of climate change, the ability of industrial forest plantation to cope with water scarcity is becoming a challenge for productivity maintenance. The phenotypic plasticity of genotypes, a major factor of adaptation to environmental changes, is still insufficiently described for long-lived species such as trees. This thesis proposes i/ to characterize the phenotypic plasticity for two commercial eucalyptus clones that differ in terms of productivity, subjected to water stress in the field, ii/ to identify potentially adaptive traits (i.e. those revealing GxE interaction) and iii/ to highlight the underlying molecular mechanisms.A field trial installed in the Republic of Congo was used to compare, responses of two contrasted eucalyptus genotypes subjected to different watering regimes during the dry season on the first two years of growth. We used an integrative approach involving different traits from gene expression to biomass production.The results show that if the effects of the dry season were relatively similar for both clones (lower relative increments), the most productive clone displayed a higher growth increment during the rainy season, and seemed to take more benefits when environmental conditions become more favorable. The ability to adjust leaf physiological, anatomical and molecular traits of this genotype seems to be an asset to maintain its photosynthetic capacity.This thesis allowed to highlight some criteria (specific leaf area, carbon isotope discrimination, thickness of collenchyma and cuticle in leaves), which could help to assess the adaptive potential of Eucalyptus populations for sustainable management of planted stands. On the other hand, some genes whose expression may be related to variation in phenotypic characters were revealed. This set of genes is resource first step toward the understanding of the molecular mechanisms underlying phenotypic plasticity in eucalyptus.