A cana-de-açúcar é uma gramínea C4 de alta biomassa que acumula grandes quantidades de sacarose e é utilizada para produção de etanol, um combustível de baixa emissão de carbono. Restrições bióticas e abióticas afetam significativamente a produtividade das culturas, pois elas podem prejudicar severamente o crescimento e desempenho da planta. Compreender as bases moleculares para essa perda de produtividade ajudará na investigação das estratégias de mitigação. Para estudar estes dois tipos de estresses, plantas jovens de cana-de-açúcar foram submetidas à herbivoria e à privação de água. Uma investigação foi realizada para estudar as mudanças transcricionais em cana-de-açúcar sujeita ao ataque da Diatraea saccharalis, usando macroarranjo para monitorar a seleção de transcritos, contendo sequências de ESTs de serina proteases e inibidores de serina proteases de cana-de-açúcar do banco de dados SUCEST. Análises do macroarranjo revelaram sequências diferencialmente expressas em resposta à herbivoria. PCR em tempo real confirmou que 10 ESTs homólogos à inibidores de protease (4 homólogos aos inibidores do tipo Bowman-Birk de arroz e milho (BBI), 5 homólogos à inibidores de proteinase de milho (MPI) e 1 homólogo ao inibidor de subtilisina) e 3 ESTs homólogos à serina proteases das famílias S1, S10 e S14 foram positivamente regulados pela herbivoria. Embora a função dos inibidores de protease na defesa está bem estabelecida, o envolvimento de proteases de planta na resposta à herbivoria ainda precisa ser elucidado. Neste trabalho nós mostramos que uma protease da família S14 foi induzida em resposta ao ataque da lagarta e ao ferimento mecânico em cana-de-açúcar. Curiosamente, sequências homólogas de arroz e Arabidopsis também responderam aos mesmos tratamentos, sugerindo um papel conservado desta protease S14 na defesa contra herbívoros. Uma importante aplicação destes resultados é a identificação de genes para utilização em estratégias biotecnológicas para melhorar a resistência da cana-de-açúcar a insetos. Outra investigação foi realizada para estudar os parâmetros fisiológicos e perfis transcricionais de genes responsivos ao estresse hídrico em plantas jovens de cana-de-açúcar. Resultados deste trabalho indicaram que a interrupção da irrigação resultou em efeitos fisiológicos mensuráveis e análises de expressão de genes selecionados de resposta ao estresse revelaram expressão diferencial significativa entre os grupos irrigado e não irrigado. Resultados do RNA-Seq revelaram atividade transcricional de 24.142 transcritos de folhas de cana-de-açúcar submetidas à seca. Análises de expressão gênica em resposta à seca revelaram 68 (resposta precoce) e 2.390 (resposta tardia) transcritos diferencialmente expressos no 3º e 7º dia de tratamento, respectivamente. O decréscimo em vários parâmetros fisiológicos foi observado depois de seis dias de privação de água. Reduções na taxa de fotossíntese, condutância estomática e transpiração foliar ocorreram antes que fossem observadas alterações físicas visíveis, mas estas foram precedidas por mudanças significativas na expressão de genes com papel na fotossíntese. RNA-Seq identificou novos transcritos com papéis na defesa precoce e tardia à seca e a validação por PCR em tempo real confirmou os resultados obtidos pelo RNA-Seq. Isto irá incentivar mais pesquisas sobre a eficiência do uso da água em cana-de-açúcar, levando a identificação de variedades com maior tolerância às condições ambientais adversas. / Sugarcane is a high biomass tropical C4 grass crop which accumulates large quantities of sucrose and is used for bioethanol production, a low-carbon emission fuel. Biotic and abiotic constraints significantly impact crop productivity, because they can severely impair plant growth and performance. Understanding the molecular basis for this loss in productivity will aid in identifying strategies for mitigation. To study these two types of stresses, young sugarcane plants were subject to herbivore and water privation. An investigation was undertaken to study the sugarcane transcriptional changes following Diatraea saccharalis damage, using macroarrays to monitor a selection of transcripts, containing sequences of sugarcane ESTs of serine proteases and serine proteinase inhibitors from the SUCEST (Sugarcane EST Project) database. Macroarray analyses revealed differently expressed sequences in response to herbivory. Real-Time PCR confirmed that 10 ESTs homologous to proteinase inhibitors (4 homologous to maize and rice Bowman-Birk inhibitors (BBI), 5 homologous to maize proteinase inhibitors (MPI) and 1 homologue to subtilisin inhibitor) and 3 ESTs homologous to serine proteases of the S1, S10 and S14 family were positively regulated by herbivory. While the protease inhibitor\'s function in defense is well established, the involvement of plant proteases in response to herbivory still remains to be elucidated. In this work we show that a sugarcane encoding S14 family protease member was upregulated in response to both D. saccharalis damage and wound treatment. Interestingly, homologous sequences from rice and Arabidopsis also responded to the same treatments, suggesting a conserved role of this S14 protease in defense against herbivores. One important application of these results is the identification of genes for use in biotechnological strategies to improve sugarcane insect resistance. Another investigation was undertaken to study the physiological parameters and transcriptional profiles of genes responsive to water stress in young sugarcane plants. Results of this work indicated that termination of irrigation resulted in measurable physiological effects in sugarcane plants and analysis of the expression of the chosen stress-response genes revealed significant differential expression between the control and treatment groups. RNA-Seq results revealed transcriptional activity of 24.142 transcripts from sugarcane leaf subjected to water stress. Gene expression analyses in response to water deprivation revealed 68 (early response) and 2,390 (later response) differentially expressed transcripts on day 3 and day 7, respectively. Sustained decreases in various physiological parameters were observed in water-stressed sugarcane plants after six days of water deprivation. Reductions in photosynthesis rate, stomatal conductance and leaf transpiration occurred before visible physical changes were observed, but this was preceded by significant changes in expression of genes with roles in photosynthesis. RNA-seq identified novel transcripts with roles in early and late response to drought stress and Real-Time qPCR validation confirmed the RNA-Seq results. This will inform further research on water use efficiency in sugarcane, leading to identification of varieties with improved tolerance to adverse environmental conditions.

Temperatura e chuva exercem grande influência sobre o crescimento, desenvolvimento e produtividade vegetal. Com o objetivo geral de verificar o efeito da variação da temperatura e chuva sobre a qualidade da fibra e a produtividade de algodão no estado de Mato Grosso, este trabalho analisou conjuntamente dados climatológicos (banco de dados contendo os valores diários de temperatura máxima, mínima e média, e chuva) e dados provenientes de um grupo de experimentos de campo de algodão. A caracterização de diversos genótipos (cultivares e linhagens) de algodão foi realizada em 109 ambientes do estado de Mato Grosso, os quais foram formados pela combinação de três causas de variação: (i) seis locais (Itiquira, Serra da Petrovina, Campo Verde, Campo Novo do Parecis, Sapezal e Sorriso); (ii) quatro épocas de semeadura (05 a 21 de dezembro, 22 de dezembro a 10 de janeiro, 11 a 25 de janeiro, 25 de janeiro a 20 de fevereiro); e (iii) oito safras agrícolas (2003/2004 a 2010/2011). Os quatro cultivares mais frequentes foram selecionados: FiberMax 966, FMT 701, FiberMax 993 e DeltaOpal, testados em 75, 68, 63 e 60 ambientes, respectivamente. Os dados climatológicos, de cada ambiente, foram segmentados em três fases de desenvolvimento do algodoeiro: (i) emergência à primeira flor (E-F1); (ii) primeira flor ao primeiro capulho, mais 21 dias (F1-C1+21D); e (iii) emergência ao primeiro capulho, mais 21 dias (F1-C1+21D). Os resultados de cada fase foram correlacionados com os dados dos experimentos de campo de algodão (produtividade de algodão em caroço, rendimento e qualidade de fibra - índice micronaire, resistência, comprimento e maturidade). Os principais resultados obtidos foram: (i) variações de temperatura e chuva ocorridas durante a fase F1-C1+21D promoveram maior influência sobre produtividade e qualidade de fibra; (ii) o aumento da amplitude térmica interferiu negativamente na produtividade de algodão em caroço, índice micronaire e maturidade de fibra, e positivamente na resistência da fibra; (iii) quando comparadas à temperatura máxima, variações de temperatura mínima foram mais limitantes sobre a produtividade e qualidade de fibra; (iv) temperaturas mínimas inferiores a 20°C interferiram negativamente na produtividade de algodão em caroço, rendimento de fibra, índice micronaire e maturidade; (v) a elevação da temperatura máxima média interferiu positivamente na resistência e negativamente no comprimento de fibra; entretanto extremos de temperatura máxima, acima de 35°C, não foram associados às variações de produtividade e qualidade de fibra; e (vi) o aumento de chuva, em quantidade acumulada e frequência de ocorrência, resultou em elevação da produtividade de algodão em caroço, índice micronaire e maturidade. Como conclusão, o aumento da temperatura mínima e da frequência de ocorrência de chuva promovem o incremento da produtividade e da qualidade da fibra de algodão, ao passo que extremos de temperatura máxima não provocam alterações consistentes no estado de Mato Grosso. / Temperature and rain have great influence on the growth, development and yield of plants. In order to verify the effect of temperature and rainfall on fiber quality and yield of cotton in the State of ´Mato Grosso´, Brazil, climatological data (daily database of maximum, minimum and mean temperature and rainfall) were analyzed with data from a group of cotton field experiments. The characterization of various genotypes (cultivars and lines) of cotton was fulfilled in 109 environments of the State of Mato Grosso, which were formed by the combination of three sources of variation: (i) six locations (´Itiquira´, ´Serra da Petrovina´, ´Campo Verde´, ´Campo Novo do Parecis´, ´Sapezal´ and ´Sorriso´); (ii) four sowing times (5 to 21 December, December 22 to January 10, 11 to 25 January, and after January 26); and (iii) eight growing seasons (2003/2004 to 2010/2011). The four most frequent cultivars were selected: ´FiberMax 966´, ´FMT 701´, ´FiberMax 993´ and ´DeltaOpal´, tested in 75, 68, 63 and 60 environments, respectively. The climatological data, from each environment, were segmented into three stages of cotton development: (i) emergence to first flower (E-F1); (ii) first flower to the first boll, plus 21 days (F1-C1+21d); and (iii) emergency to the first boll, plus 21 days (E-C1+21d). The results of each phase were correlated with the experimental field data of cotton (productivity, lint yield and fiber quality (micronaire index, strength, length and maturity). The main results were: (i) variations of temperature and rainfall during the phase (F1-C1+21d) promoted greater influence on yield and fiber quality; (ii) the increasing of thermic amplitude affected negatively on yield, micronaire index and fiber maturity, and positively the fiber strength; (iii) when compared to the maximum temperature, minimum temperature variations were more restrictive on yield and fiber quality; (iv) values of minimum temperature below 20°C were harmful to yield, lint yield, micronaire index and maturity; (v) the maximum temperature had a positive influence in resistance and a negative influence on the fiber length; however, extreme maximum temperature above 35°C wasn\'t associated to variations on yield and fiber quality; and (vi) the increasing of rainfall, in height and occurrence frequency, influenced the rising in cotton productivity, micronaire index and maturity. In conclusion, the increasing of minimum temperature and frequency of rainfall occurrence promote the increment of cotton productivity and fiber quality, while maximum temperature extremes caused no consistent changes in the State of Mato Grosso.

Costus arabicus L. é uma espécie herbácea, nativa da Mata Atlântica, conhecida popularmente como costus, canela-de-ema, cana-do-brejo, cana-de-macaco e gengibre-espiral, que é comumente utilizada como planta ornamental e na medicina popular. O órgão subterrâneo (rizoma) dessa espécie acumula amido como principal carboidrato de reserva. O estresse hídrico por falta de água é um dos fatores ambientais mais importantes que podem regular o crescimento e desenvolvimento das plantas, limitando sua produção e levando também a alterações nas características fisiológicas, bioquímicas e anatômicas. Desse modo, a deficiência hídrica destaca-se como fator adverso ao crescimento e à produção vegetal por alterar a eficiência com que os fotoassimilados são convertidos para o crescimento e desenvolvimento das plantas. O objetivo deste trabalho foi avaliar a influência do déficit hídrico no crescimento e acúmulo de carboidratos de reserva em plantas de Costus arabicus L., assim como na anatomia e ultra-estrutura do rizoma. O déficit hídrico foi imposto às plantas em duas épocas diferentes (inverno e verão) através dos seguintes tratamentos: plantas irrigadas diariamente (controle) e irrigadas a cada 7 (7d) e 15 dias (15d). Foram avaliados a umidade do solo, o teor relativo de água nas folhas (TRA), o potencial hídrico foliar (w), o potencial osmótico da seiva do rizoma (s), crescimento vegetativo, teores dos açúcares solúveis totais (AST) e redutores (AR), amido, aminoácidos livres totais e prolina, assim como, aspectos anatômicos e ultra-estruturais do rizoma. A umidade do solo manteve-se praticamente constante nas plantas irrigadas diariamente e superior a das plantas irrigadas a cada 7 e 15 dias nos dois experimentos. O déficit hídrico afetou o crescimento de Costus arabicus e os menores valores de w e s das plantas tratadas coincidiu com os menores valores de massa seca e número total de folhas verdes e aumentos do número de folhas senescidas. O baixo teor de prolina presente nas plantas sob déficit hídrico sugere que esse aminoácido não atuou na osmorregulação dessas plantas. Os menores teores de AST, AR e a manutenção dos teores de amido nos rizomas, associados ao menor crescimento das plantas dos tratamentos 7d e 15d em relação às plantas controle, sugerem a ocorrência de dormência do órgão subterrâneo em resposta ao déficit hídrico imposto, evidenciando uma estratégia de evitação à seca. As análises anatômicas mostraram que o rizoma é revestido pela epiderme ou súber estratificado e que existe uma delimitação entre as regiões cortical e vascular. Também foi observada a presença de numerosos grãos de amido e de idioblastos com conteúdo fenólico no parênquima dessas duas regiões. Entre os tratamentos, os rizomas das plantas irrigadas a cada 7 dias apresentaram características anatômicas semelhantes aos das plantas controle. No entanto, nos rizomas das plantas irrigadas a cada 15 dias, as células do tecido de revestimento e das camadas corticais mais externas tornaram-se mais achatadas e com paredes sinuosas. Através das análises em microscópio eletrônico de varredura, observou-se que os grãos de amido das plantas irrigadas diariamente e daquelas submetidas ao déficit hídrico apresentaram os mesmos formatos. / Costus arabicus L. is an herbaceous species, native to the Atlantic Forest, known popularly as costus, canela-de-ema, cana-do-brejo, cana-de-macaco and gengibreespiral, that is commonly used as an ornamental plant and in popular medicine. The underground organ (rhizome) of this species accumulates starch as the main reserve carbohydrate. Water stress, due to low water availability, is one of the most important environmental factors that regulates plant growth and development, limiting the production and leading to changes in physiological, biochemical and anatomical features. Thus, water deficit stands out as a factor adverse to growth and crop production by altering the efficiency by which photoassimilates are converted for growth and development of plants. The aim of this study was to evaluate the effect of water deficit on growth and accumulation of reserve carbohydrates in plants of Costus arabicus L., as well as on the anatomy and ultrastructure of the rhizome. Water deficit was imposed on plants in two seasons (winter and summer) through the following treatments: daily irrigation (control) and irrigation every 7 (7d) and 15 days (15d). We evaluated soil moisture, relative water content in leaves (RWC), leaf water potential (w), osmotic potential of sap from the rhizome (o), vegetative growth, levels of total soluble sugars (TSS), reducing sugars (RS), starch, amino acids and proline, as well as anatomical and ultrastructural characteristics of the rhizome. Soil moisture remained mostly constant in irrigated plants and higher than that of plants irrigated every 7 to 15 days in both experiments. Water deficit affected plant growth; low values of w and o of treated plants coincided with the low values of dry weight and total number of green leaves and with increases in the number of senesced leaves. The low content of proline found in plants under water deficit suggests that this amino acid did not act in the osmoregulation of these plants. The lowest levels of TSS, RS and the maintenance of starch levels in rhizomes, associated with low rates of plant growth in treatments 7d and 15d, compared to control plants suggest the occurrence of dormancy of the underground organ in response to water deficit, indicating a strategy of drought avoidance. The anatomical analysis showed that the rhizome is covered by epidermis or stratified cork and there is a delimitation between the cortical and vascular regions. We detected the presence of numerous starch grains and idioblasts with phenolic content in the parenchyma of these two regions. The rhizomes of plants irrigated every 7 days showed anatomical features similar to control plants. However, in the rhizomes of plants irrigated every 15 days, the cells of covering tissue and of the outer cortical layers became more flattened and sinuous. Analysis of scanning electron microscopy showed that the starch grains of irrigated plants and of plants submitted to drought presented the same shape.

Para que ocorra a indução floral em citros, as plantas necessitam passar por algum tipo de estresse hídrico ou térmico (baixas temperaturas). Apesar do estresse hídrico ser importante para o florescimento, condições extremas deste tipo de estresse podem prejudicar o desenvolvimento e a fixação dos frutos na planta posteriormente ao período de florescimento. Neste sentido, a redução da fração de área molhada do solo pode também ser uma fonte de estresses hídrico nas plantas. Dessa forma, o presente trabalho tem como objetivo estudar o efeito do estresse hídrico e da fração de área molhada (100% e 12,5%) nas relações hídricas da laranjeira Valência, sob dois tipos solo e dois porta-enxertos. O experimento foi conduzido na área de pesquisa do Departamento de Engenharia de Biossistemas na Escola Superior de Agricultura Luiz de Queiroz (ESALQ/USP) em ambiente protegido. Foi utilizado o delineamento em esquema fatorial 2 x 2 x 2 x 2 com os tratamentos dispostos em faixas, totalizando 16 tratamentos, constituídos da combinação de dois tipos de solos (argiloso e franco-arenoso), dois porta-enxertos (limoeiro Cravo e citrumelo Swingle), duas frações de área molhada (100% e 12,5%) e dois níveis de deficiência hídrica: 1) Estresse Hídrico Contínuo (suspensão da irrigação por 30 dias) 2) Estresse Hídrico Intermitente (sub-lâmina) - aplicação de 30% da ET0. As plantas foram conduzidas em caixas de 500 L internamente divididas em compartimentos. O inicio do experimento de estresse hídrico consistiu em realizar a irrigação do volume total do solo, elevando-o a capacidade de campo, sendo suspensas as irrigações durante o período de avaliação, sendo após realizado o acompanhamento diário do consumo hídrico em cada compartimento das caixas. Amostrou-se o sistema radicular das plantas, para cada compartimento existente para averiguar a existência de adaptação radicular em função da restrição de área molhada. Determinou-se simultaneamente a transpiração de todas as plantas através de sondas de dissipação térmica, o conteúdo de água no solo, o crescimento das plantas (área foliar e diâmetro de caule), o potencial de água na folha e a temperatura foliar. A extração de água no solo após 6 meses de irrigação localizada indica que o sistema radicular na zona irrigada consegue redistribuir parte da água absorvida, pelo sistema radicular da planta que se encontra na área seca, mantendo-o vivo. Observa-se que no solo argiloso as plantas apresentaram um maior comprimento total de raízes do que no solo arenoso. A média do módulo do potencial da água na folha das plantas sob condição simulada de irrigação por gotejamento, foi 31,5% maior quando comparado às plantas com 100% de solo molhado (-1,085 MPa e -0,7435 MPa). Em termos de área foliar, o estresse contínuo promoveu uma maior desfolha das plantas, sendo significativa a diferença entre os tipos de solo, as condições de estresses e a interação tipo de solo e porta-enxertos. Treze dias após a imposição dos estresses, a temperatura foliar, nos períodos de maior demanda atmosférica (14:00hs) tende a ser maior do que a temperatura do ar. Independente do tipo de estresse, para solo arenoso a transpiração diminui quanto se impõe a redução de área molhada (gotejamento). Esperava-se que os tipos de estresse hídrico impostos apresentassem uma diferenciação de consumo hídrico nas plantas, no período de retomada da irrigação, sendo o estresse intermitente o que deveria apresentar o maior consumo hídrico; nos resultados obtidos não ficou muito clara e estabilizada esta tendência, apesar do nível de desfolha e o potencial de água nas folhas confirmaram as hipóteses inicialmente formuladas. Existe a possibilidade de que o método de dissipação térmica tenha sofrido influência significativa da desfolha no gradiente térmico natural, o que pode ter ocasionado imprecisão nos dados coletados e resultado inconsistência nos fluxos de seiva calculados neste período de baixo índice de área foliar. / In order to present floral induction, citrus plants need to undergo some type of thermal (low temperature) or water stresses. However, despite a certain level of water stress is important for the flowering, excessive water stress can impair the development and fruit set in the plant. In this sense, to reduce the fraction of wetted area of the soil can also be a source of water stress in plants. This paper aims to study the effect of water stress and the fraction of wetted area (100% and 12.5%) in Valencia orange tree transpiration under two soil types and two rootstocks. The experiment was conducted at the Department of Biosystems Engineering at the School of Agriculture \"Luiz de Queiroz\" (ESALQ / USP) in a protected environment. It was used a randomized design in a factorial 2 x 2 x 2 x 2 with treatments arranged in strips, totaling 16 treatments combinations, consisting of two soil types (clay and sandy loam), two-door grafts (Rangpur lime and citrumelo \'Swingle\'), two fractions of wetted area (100% and 12,5%) and two levels of water stress: 1) Continuous Stress (suspension of irrigation for 30 days) 2) Intermittent Stress (deficit irrigation: 30% ET0). The plants were conducted in 500 L boxes divided into compartments inside. At the beginning of the water stress experiment, soil moisture was set to field capacity. Irrigation was suspended during the evaluation period, and it was monitoring every day the water consumption in each compartment boxes. Root systems of plants were sampled for each compartment to determine total length of roots in the soil. It were determined the individual transpiration of each plant using thermal dissipation probes (sap flow / Granier), the water content in soil, the plant growth (leaf area and stem diameter), the leaf water potential and leaf temperature. The extraction of water in the soil after 6 months of drip irrigation indicates that the root system in the irrigated area redistribute some of the absorbed water to the root system that is in the dry area, keeping it alive waiting for the raining period. It was observed for the clay soil a greater total length of roots compared to the sandy soil. The mean of leaf water potential for plants under simulated drip irrigation conditions (12.5 % wetted area) was 31,5% higher compared to plants with 100% of wetted area (-1,085 MPa and -0,7435 MPa). In terms of leaf area, the continuous stress promoted a greater leaf loss of plants, with a significant difference between soil types, stress types and the interaction of soil type and rootstock. Thirteen days after the imposition of the water stresses, leaf temperature at around 2:00 pm tends to be higher than air temperature. For sandy soil, independent of the water stress type, transpiration decreases as the wetted area is reduced (drip). It was expected for the water stress types imposed, a differentiation of plants water consumption after resuming irrigation for fruit set, being the intermittent stress the greatest water consumption treatment. The results obtained are unclear and non-stabilized for this trend, although leaf losses and leaf water potential confirmed the hypothesis initially formulated. There is a possibility that thermal dissipation probles suffered significant influence of leaf losses on the natural thermal gradient, which may have caused inaccuracies in the collected data, resulting inconsistent values of sap flow calculated for this period of low leaf area index.

Made available in DSpace on 2014-08-20T13:59:12Z (GMT). No. of bitstreams: 1 dissertacao_marilia_carvalho_carneiro.pdf: 420554 bytes, checksum: 1d98e1f75981deb53d68fc556ba8870a (MD5) Previous issue date: 2011-02-25 / This study aimed evaluates parameters related to photosynthesis and antioxidant metabolism in two sunflower cultivars under different water regimes in the initial growth phase. Seeds of the cultivars MG2 and M735 were grown in pots with a capacity of 10 L, using soil as substrate. Thirty days after sowing was done a thinned to four plants per pot, watered daily. Two months after the treatments were established, where a portion of the plants of each cultivar was kept watered daily (control) and another submitted to the full suspension of irrigation (water deficit). After 24 hours, five and twelve days of induction treatment in predawn leaf water potential was measured and leaf chlorophyll content in the early hours. Between 9:00 and 10:00 hours were measured photosynthetic parameters and chlorophyll fluorescence. Then, leaves and roots were collected and stored in a freezer at -80°C for quantification of cell damage and antioxidant enzymes. In response to water stress imposed at the end ofof the evaluations , there was a sharp drop in water potential of both cultivars . On the other hand, the chlorophyll index was down only in cv M735. The decline in leaf water potential reflected in reductions in stomatal conductance, transpiration rate, net assimilation of CO2 and its internal concentration in both cultivars, emphasizing greater reductions in transpiration and internal CO2 concentration, hence increasing efficiency water use for both varieties. The fluorescence of chlorophyll a decreased effective quantum yield photochemistry (FV '/ FM') and produce effective quantum of photosystem II (ΦPS2) in cv MG2. During that same period we observed an increase in the levels of lipid peroxidation, H2O2 content and activity of SOD and CAT, with the greatest difference compared to control, especially in cv M735, which prevented more effectively oxidative stress. / O presente estudo teve por objetivo avaliar parâmetros relacionados à fotossíntese e metabolismo antioxidante em duas cultivares de girassol submetidas a diferentes regimes hídricos, na fase inicial de crescimento. Sementes das cultivares MG2 e M735 foram semeadas em vasos com capacidade para 10 L, utilizando solo como substrato. Trinta dias após a semeadura foi realizado um desbaste mantendo quatro plantas por vaso, irrigadas diariamente. Após dois meses foram estabelecidos os tratamentos, onde uma parcela das plantas de cada cultivar foi mantida irrigada diariamente (controle) e a outra submetida à suspensão total da irrigação (déficit hídrico). Após 24 horas, cinco e doze dias da indução dos tratamentos, na antemanhã foi avaliado o potencial hídrico foliar e nas primeiras horas seguintes o índice de clorofila. Entre 9:00 e 10:00 horas foram mensurados parâmetros fotossintéticos e fluorescência da clorofila a. A seguir, folhas e raízes foram coletadas e armazenadas em freezer a - 80oC para quantificação dos danos celulares e atividade de enzimas antioxidantes. Em resposta ao estresse hídrico imposto, ao final das avaliações, observou-se uma queda acentuada no potencial hídrico das duas cultivares. Por outro lado, o índice de clorofila sofreu queda somente na cv M735. A queda no potencial de água nas folhas refletiu em reduções na condutância estomática, taxa transpiratória, assimilação líquida do CO2 e na sua concentração interna para as duas cultivares, destacando maiores reduções na transpiração e concentração interna de CO2, o que justifica o aumento da eficiência no uso da água para ambas as cultivares. A fluorescência da clorofila a apresentou queda na eficiência quântica fotoquímica efetiva (FV /FM ) e na produção quântica efetiva do FS II (ΦPS2) nas plantas da cv MG2. Nesse mesmo período foi observado aumento nos níveis de peroxidação de lipídios, teores de H2O2 e na atividade das enzimas SOD e CAT, com maior diferença em relação ao controle, principalmente, na cv M735, que evitou de forma mais efetiva o estresse oxidativo.

La sélection des nouveaux porte-greffes du genre Prunus a pour principal objectif d’utiliser lavariabilité génétique existant au sein des différentes espèces de Prunus afin de créer un matérielvégétal innovant, performant au plan agronomique et résistant à différentes contraintes biotiques etabiotiques. L’exploitation de la variabilité génétique s'appuie sur le phénotypage des différentsindividus présents dans les collections de ressources génétiques pour les caractères recherchés. Celapermet de sélectionner des génotypes spécifiques pour améliorer un trait donné.Dans cette étude constituée de deux parties, nous avons exploré la diversité génétique au seindu genre Prunus pour ce qui concerne deux traits importants : l’aptitude au bouturage et les réponsesde certains porte-greffes à la contrainte hydrique.Dans la première partie l’aptitude au bouturage ligneux a été explorée dans une collectiongénétique de 222 génotypes. Les résultats obtenus montrent une variabilité considérable entre les sousgenresAmygdalus et Prunophora et également une variabilité interspécifique au sein de chaque sousgenre.La réussite au bouturage est nettement améliorée chez les hybrides interspécifiques dont un desparents appartient à l’espèce P cerasifera.Dans la deuxième partie nous avons étudié les réponses à la contrainte hydrique chez neufgénotypes couramment utilisés comme porte-greffes. Nous avons comparé trois régimes hydriquesdifférents : témoins, stressés et recouvrés. L’humidité du substrat est maintenue à la capacité au champtout au long de l’expérience pour les plants témoins, par contre l’arrosage a été arrêté pendant 14 jourspour les plants stressés, puis il a été repris pendant 10 jours pour les plants recouvrés. Durant cetteexpérience, nous avons effectué des mesures morphologiques et physiologiques sur la partie aérienneainsi que des excavations à la fin de chaque phase pour examiner les modifications au niveau dusystème racinaire. Sur la partie aérienne, la contrainte hydrique a provoqué une diminutionsignificative de la photosynthèse nette, de la transpiration totale, de la conductance stomatique, et del’expansion des feuilles.Le système racinaire a répondu à cette contrainte par plusieurs modifications. D’abord, le ratio racines/pousses a augmenté pour 4 génotypes (GF305, GF677, Montclar et Myrobolan1254). Ensuite, lesdifférents traits de l’architecture racinaire ont montré des modifications sous l’effet de la contraintehydrique : la longueur de la zone apicale non ramifiée (LZANR) qui traduit l’élongation racinaire,ainsi que les diamètres apicaux des racines ont diminué chez tous les génotypes. Par conséquent lesracines se sont affinées et ont réduit leur croissance en longueur. Produire des racines plus finesaugmente la surface de contact entre les racines et le sol et améliore la capacité d’absorption. Unediminution de la distance inter-ramification a été observée chez les plants stressés. Cette modificationpourrait s’expliquer par le fait que les plantes produisent plus de racines latérales en profondeur oùl’eau est souvent plus disponible. De plus, les racines latérales produites étaient également plus fines.Au plan qualitatif, les génotypes ont eu des réponses semblables, mais l’intensité de la réponse a variéselon les génotypes. / In breeding programs of Prunus rootstocks the aim is to use the existing genetic variabilitywithin Prunus species in order to create new rootstock genotypes with excellent agronomic traits, andimproved resistance to biotic and abiotic stresses.Exploitation of the genetic variability is based on the evaluation of phenotypic variation amongindividuals within genetic collections for desirable traits. This make possible to select specificgenotypes to improve a given trait.This study consists of two parts; we explored the genetic diversity within the genus Prunusregarding two important characteristics: rooting ability of hardwood cuttings and responses of somerootstocks to water stress.In the first part, rooting ability of hardwood cuttings was evaluated among 222 genotypespreserved in genetic collection. The results show considerable variability among the sub genusAmygdalus, Prunophora, and also an interspecific variability within each of them. Rooting ability byhardwood cutting was significantly improved in interspecific hybrids if one parent belongs to Pcerasifera species.In the second part of this study we studied the responses of nine genotypes, commonly usedrootstocks, to water stress. We applied three treatments: control, water stress and recovering.Soil moisture was maintained at field capacity through all stages of the experiment for the controlplants. On the contrary we stopped watering during 14 days for the stressed plants, and then we rewateredthe recovered plants for 10 days.During this experience, we performed morphological and physiological measurements on the aboveground parts of plants and we excavated plants at the end of each phase to observe root systemmodifications.Aboveground parts of plant responded to water stress by a significant decrease in net photosynthesis,total transpiration, stomatal conductance and leaf expansion.Root system responded to water stress by several modifications:Four genotypes (GF305, Montclar, GF677 and Myrobolan) showed a significant increase in root toshoot ratio under drought conditions. We also detected morphological modifications on the differenttraits of root architecture in response to water stress.The length of the apical unbranched zone LAUZ and the apical diameter were decreased forall genotypes, consequently, roots became finer and reduced their rate of elongation. Fine rootsenhance the surface of contact between roots and soil which in turn improve the acquisition of waterunder drought condition.The inter-branch distance also responded, and it tended to decrease under the water stress treatment.The decrease in inter-branch distance can be explained by a production of more lateral roots in deeplayers, where water was more available, moreover these new laterals roots were also finer.Qualitatively, a common response to water stress was observed on the different traits of the rootsystem architecture, but we showed a genotypic effect determining the level of the response.

Le sujet de l’adaptation aux changements climatiques est devenu l’un des sujets contemporains les plus importants dans la vigne. Une grande focalisation a été mise sur la compréhension des effets du porte-greffe sur la croissance du scion, l’absorption des nutriments, et la tolérance au stress, dans l’objectif final de développer de nouveaux porte-greffes qui facilitent l’adaptation au changement climatique. L’objectif de cette thèse est d’examiner comment les différences dans la résistance à la sécheresse entre les génotypes peut résulter en de grandes différences dans leur capacité à maintenir leur croissance racinaire en situation de stress. Une meilleure compréhension sur la manière dont la structure, la croissance racinaire et l’absorption d’eau répondent au stress nous permettra de mieux comprendre quels sont les aspects de la physiologie racinaire qui contribuent à la tolérance face à la sécheresse. Des recherches précédentes qui s’étaient focalisées sur l’absorption d’eau racinaire chez la vigne ont suggéré que l’absorption d’eau racinaire pouvait être fortement liée à la vitesse de croissance racinaire instantanée (voir Gambetta et al. 2013). Cette observation implique que des différences entre les génotypes dans la résistance face à la sécheresse pourrait largement résulter de leur capacité à maintenir la croissance racinaire en conditions de stress. Deux porte-greffes de vigne avec des capacités contrastées en matière de résistance à la sécheresse, le Riparia Gloire de Montpellier (RGM) et le 110 Richter (110R) ont été sélectionnés pour étudier dans cette thèse. RGM est considéré comme sensible à la sécheresse, tandis que 110R est fortement résistant à la sécheresse (Carbonneau 1985). La thèse a examiné la relation entre la croissance racinaire et la capacité de résistance à la sécheresse en évaluant la vitesse de croissance racinaire, la conductivité hydraulique à travers deux variétés de porte-greffe en conditions de déficit en eau. Le niveau de l’expression des gènes d’aquaporines (via la qPCR et l’ARNseq) et leur contribution à la conductivité hydraulique racinaire ont été analysés dans les radicelles afin d’obtenir une meilleure compréhension sur les mécanismes impliqués dans la régulation de l’absorption de l’eau racinaire et la conductivité hydraulique au cours du développement et en réponse à un manque d’eau.Le traitement de stress d’eau prolongé a diminué le potentiel hydraulique de la plante. La croissance racinaire individuelle est très hétérogène : bien que le traitement de sécheresse réduise l’élongation racinaire en moyenne, la vitesse de croissance racinaire varie tout de même énormément. Un haut niveau de stress hydrique a réduit significativement la vitesse de croissance racinaire moyenne à la fois pour RGM et 110R. Globalement, la vitesse de croissance racinaire moyenne a montré une tendance réduite au cours du développement de la plante. La température du sol est aussi un facteur qui affecte la croissance racinaire. Pour RGM et 110R, en conditionsIIde bon arrosage et de stress hydrique, la vitesse de croissance quotidienne moyenne a été positivement corrélée avec la température du sol quotidienne moyenne. En conditions de bon arrosage, des vitesses de croissance racinaires plus importantes ont été constamment observées chez 110R par rapport à RGM, ce qui pourrait être une explication possible de sa meilleure résistance à la sécheresse par rapport à 110R. [...] / The subject of adaptation to climate change has become one of the most important contemporary topics in grapevine. Much focus has been placed on the understanding of rootstocks effects on scion growth, nutrient uptake, and tolerance to stress, with the ultimate goal of developing novel rootstocks that facilitate adaptation to a changing climate. The purpose of this thesis is to examine how differences in drought resistance between genotypes could result largely from differences in their ability to maintain root growth under stress. A better understanding of how root structure, growth, and water uptake respond to stress will allow us to better understand what aspects of root physiology contribute to drought tolerance. Previous research focused on root water uptake in grapevine suggested that root water uptake could be tightly coupled to a root’s instantaneous rate of growth (see Gambetta et al. 2013). This observation implies that differences in drought resistance between genotypes could result largely from their ability to maintain root growth under stress. Two grapevine rootstocks with contrasting drought resistance capacity, Riparia Gloire de Montpellier (RGM) and 110 Richter (110R), were selected to study in this thesis. RGM is considered as sensitive to drought, while 110R is highly resistant to drought (Carbonneau 1985). The thesis examined the relationship between root growth and drought resistant capacity by assessing root growth rate, hydraulic conductivity across two rootstock varieties subjected to water deficit. The role of aquaporin gene expression (via qPCR and RNAseq) and their contribution to root hydraulic conductivity were analyzed in fine roots in order to obtain a better understanding on the mechanisms involved in the regulation of root water uptake and hydraulic conductivity across development and in response to water deficit.Prolonged water stress treatment decreased plant water potential. Individual root growth is very heterogeneous, although drought treatment reduces root elongation on average, individual root growth rate still varies enormously. High level of water stress significantly reduced average root growth rate for both RGM and 110R. Globally, average root growth rate showed a decreased trend over plant development. Soil temperature is also a factor that affects root growth. For both RGM and 110R, under both well-watered and water-stressed conditions, average daily root growth rate was positively correlated with average daily soil temperature. Under well-watered conditions, higher root growth rates were constantly observed in 110R compared to RGM, which could be one possible explanation for the higher capacity in drought resistance of 110R.Root hydraulic conductivity (Lpr) was influenced by both water stress treatment and plant developmental stage. Generally, for both RGM and 110R, Lpr was significantly reduced under water stress in early stage. In mid and late stages, no significant differences in Lpr were observedIVbetween well-watered and water-stressed plants. Changes in individual root Lpr in response to pre-dawn leaf water potential (ᴪpredawn) were investigated as well. Lpr showed a fast drop in the beginning of water stress treatment when ᴪpredawn was higher than -0.5 MPa. However, with ᴪpredawn getting more negative, e.g. from -0.4 MPa to -2.0 MPa, the range of Lpr values measured in our study maintained constant. Lpr of well-watered plants decreased as well even though their ᴪpredawn was maintained at a high level (< 0.1 MPa) during the period of the experiment. [...]

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Wheat is the main winter crop in Rio Grande do Sul State. Temperature and photoperiod are the major environmental factors that control development in wheat. Wheat plants exposed to cold, non freezing temperatures induce them to enter the reproductive phase. Temperature and photoperiod response functions are used to simulate the wheat development. Developmental models are important part of water soil balance, growth and yield crop models. Thus, the objectives of this thesis were: (i) to evaluate vernalization sensitivity of some Brazilian wheat genotypes and to fit a vernalization response function for these genotypes; (ii) to determine the best temperature, photoperiod and vernalization functions to simulate wheat development with the Wang and Engel (WE) model for some Brazilian wheat genotypes; and (iii) to improve the prediction of available soil water in the soil by modifying two mechanistic models. To achieve the first objective, a factorial experiment with eight levels of vernalization (0, 7, 14, 21, 28, 35, 42 and 49 vernalization days) and six wheat genotypes were used. Also a field experiment was carried out in Santa Maria, RS, Brazil, to provide independent data on the date of developmental stages of six Brazilian wheat genotypes for evaluating the original and the modified WE model. Soil water content was determined with the gravimetric method to evaluate water balance models during three sowing dates. For those genotypes that presented some response to vernalization, response functions are presented, which can be used in models for simulating the development of Brazilian wheat cultivars with distinct vernalization requirements. The WE model simulates better the development of the Brazilian wheat genotypes using cardinal temperatures higher than original WE model and using daily maximum and minimum temperatures in the calculation of the temperature response function. The modified Campbell; Diaz water balance model is more realistic and has a greater potential for performing well in regions others than it was developed. The modified Amir; Sinclair water balance model simulates better the available soil water than the original model. / O trigo é a principal cultura de inverno do Estado do Rio Grande do Sul. A temperatura e o fotoperíodo são os dois elementos meteorológicos que mais influenciam o desenvolvimento de plantas de trigo. Quando se expõem plantas de trigo a temperaturas baixas não congelantes, estas podem ser induzidas a entrarem no estágio reprodutivo, diminuindo o seu ciclo de desenvolvimento. Para simular o desenvolvimento do trigo são utilizadas funções de resposta a temperatura e fotoperíodo. O modelo de desenvolvimento é parte importante em modelos de simulação de balanço de água no solo, crescimento e rendimento das culturas. Assim, os objetivos desta tese foram: (i) determinar a sensibilidade à vernalização de alguns genótipos de trigo do sul do Brasil e, ajustar uma função de reposta a vernalização para aqueles que apresentaram sensibilidade; (ii) determinar a melhor função de temperatura, fotoperíodo e vernalização para simular o desenvolvimento de alguns genótipos brasileiros de trigo com o modelo de Wang e Engel (WE); e (iii) obter uma melhor predição do balanço de água no solo, modificando dois modelos mecanísticos de balanço de água no solo. Para atingir o primeiro objetivo, foi conduzido um experimento fatorial com oito níveis de vernalização (0, 7, 14, 21, 28, 35, 42 e 49 dias de vernalização) e seis genótipos de trigo. Também foi realizado um experimento com várias datas de semeadura em Santa Maria, RS, Brasil, para obtenção de dados independentes dos estágios de desenvolvimento de seis genótipos Brasileiros de trigo para avaliar o modelo WE modificado. Foi determinado o conteúdo de água no solo pelo método gravimétrico em três datas de semeadura, com a finalidade de avaliar o desempenho dos dois modelos de balanço de água no solo. Para os genótipos que apresentaram resposta à vernalização são apresentadas funções de resposta, que podem ser usadas em modelos de simulação do desenvolvimento de genótipos brasileiros de trigo. O modelo WE simula melhor o desenvolvimento do trigo quando são usadas as temperaturas cardinais maiores do que as do modelo original e usando as temperaturas mínimas e máximas diária para o cálculo da função de resposta a temperatura. O modelo de balanço de água no solo de Campbell; Diaz modificado é mais realístico e com maior possibilidade de desempenho satisfatório em regiões de clima distinto daquelas em que foi desenvolvido. O modelo de balanço de água no solo de Amir; Sinclair modificado estima melhor a água disponível no solo que o modelo original.

Dans le cadre du changement climatique et de l'évolution de la réglementation concernant les intrants azotés, la culture du colza risque d'être fortement pénalisée dans la mesure où c'est une culture qui nécessite d'importants apports azotés pour atteindre son potentiel de rendement. Par ailleurs, comme chez le colza un déficit hydrique induit l'accumulation de certains composés azotés, il est vraisemblable que cela conduise au détournement d'une quantité importante d'azote vers les organes végétatifs aux dépens des organes reproducteurs et donc du rendement. Chez le colza, la réponse métabolique au déficit hydrique se traduit par une très forte accumulation de proline et dans une moindre mesure une augmentation de la teneur en GABA (acide γ-aminobutyrique), deux acides aminés connus chez la plupart des plantes pour leur réponse à de nombreux stress abiotiques. L'objectif de cette thèse est de déterminer comment le métabolisme de ces deux molécules contribue à l'allocation de l'azote au cours du développement de la plante en situation normale comme en condition de stress hydrique et/ou azoté. Pour répondre à cette question nous avons fait le choix de caractériser deux voies enzymatiques majeures impliquées dans le catabolisme de la proline et du GABA : la proline déshydrogénase (ProDH) et la GABA-transaminase (GABA-T) et d'évaluer l'impact de carences hydriques et/ou azotées sur ces voies. Cette étude nécessitait d'identifier au préalable les gènes codant ces enzymes afin d'aborder une approche fonctionnelle. Les résultats montrent l'existence de multiples copies de gènes ProDH et GABA-T dans le génome du colza. L'analyse de leurs profils d'expression suggère que des processus de sub-fonctionnalisation sont en cours conduisant à l'expression spécifique, de certaines copies en réponse aux stress, et d'autres dans les processus développementaux. La comparaison des profils métaboliques avec les profils spécifiques des transcrits a permis d'élaborer des hypothèses sur le rôle de ces voies dans la gestion de l'azote. L'étude conjointe des métabolismes de la proline et du GABA suggère l'existence de régulations connexes entre les deux. Enfin, l'utilisation de plantules a permis - d'approfondir la régulation des gènes étudiés à des stades précoces de développement - et de mettre en évidence les effets délétères de l'inhibition de la GABA-T par une approche pharmacologique. En conclusion ces résultats apportent des précisions sur la régulation de ces deux enzymes et fournissent des éléments de réponse quant au rôle fonctionnel des catabolismes de la proline et du GABA dans les processus de gestion de l'eau et de l'azote chez le colza. Ces travaux constituent donc une première étape dans une démarche de validation de ces gènes comme candidats pour des programmes d'amélioration du colza visant à sélectionner des génotypes mieux adaptés aux conditions environnementales futures. / In the context of climate change and recent regulation concerning nitrogen inputs, the oilseed rape yields may be severely decreased because its crop requires significant nitrogen supply to reach high yield performance. Moreover, as water deficit induces the accumulation of some nitrogen compounds in oilseed rape, it is likely that this could lead to diversion of significant amounts of nitrogen to the vegetative organs at the expense of the reproductive ones and therefore of the yield. In oilseed rape, the metabolic response to water deficit results in a very high proline accumulation and, to a lesser extent, an increased content of GABA (γ-aminobutyric acid), both these amino acids known for their response to many environmental stresses in most species. The objective of the work presented here was to determine how the metabolism of proline and GABA contributes to the nitrogen allocation during plant development under optimal conditions and under water stress and/or nitrogen depletion. To answer this question, we have chosen to characterize two major enzymatic pathways involved in the catabolism of proline and GABA, proline dehydrogenase (ProDH) and GABA transaminase (GABA-T), and assess the impact of water and/or nitrogen deficiency on these pathways. This study has required to preliminary identify the genes encoding these enzymes in order to initiate a functional approach. The results show the presence of multiple copies of ProDH and GABA-T genes in the oilseed rape genome. Analysis of their expression profiles suggests that sub-functionalization processes are occurring, leading to the specific expression of some copies in response to stress, and some in developmental processes. Comparison of metabolic profiles with specific profiles of transcripts allows us to hypothesize about the role of these pathways in management of nitrogen. The combined study of proline and GABA metabolisms suggests the existence of relationships between them. Finally, the use of seedlings allows - further studying the regulation of genes in the early stages of development - and highlighting the deleterious effects of the inhibition of GABA-T by a pharmacological approach. In conclusion these results supply information on the regulation of these two enzymes and provide answers about the functional roles of proline and GABA catabolisms in the management processes of water and nitrogen in oilseed rape. These works constitute a first step in validation process of these genes as putative candidates for oilseed rape breeding programs to select genotypes better adapted to future environmental conditions.

Traditional irrigation was practiced in Ethiopia since time immemorial. Despite this, water productivity in the sector remained low. A survey on the Godino irrigation scheme revealed that farmers used the same amount of water and intervals, regardless of crop species and growth stage. In an effort to improve the water productivity, two traditional irrigation scheduling methods were compared with two scientific methods, using furrow irrigation. The growth performance and tuber yield of potato (cv. Awash) revealed that irrigation scheduling using a neutron probe significantly outperformed the traditional methods, followed by the SWB model Irrigation Calendar. Since the NP method involves high initial cost and skills, the use of the SWB Calendar is suggested as replacement for the traditional methods. SWB is a generic crop growth model that requires parameters specific to each crop, to be determined experimentally before it could be used for irrigation scheduling. It also accurately describes deficit irrigation strategies where water supply is limited. Field trials to evaluate four potato cultivars for growth performance and assimilate partitioning, and onions' critical growth stages to water stress were conducted. Crop-specific parameters were also generated. Potato and onion crops are widely grown at the Godino scheme where water scarcity is a major constraint. These crop-specific parameters were used to calibrate and evaluate SWB model simulations. Results revealed that SWB model simulations for Top dry matter (TDM), Harvestable dry matter (HDM), Leaf area index (LAI), soil water deficit (SWD) and Fractional interception (FI) fitted well with measured data, with a high degree of statistical accuracy. The response of onions to water stress showed that bulb development (70-110 DATP) and bulb maturity (110-145) stages were most critical to water stress, which resulted in a significant reduction in onion growth and bulb yields. SWB also showed that onion yield was most sensitive to water stress during these two stages. An irrigation calendar, using the SWB model, was developed for five different schemes in Ethiopia, using long-term weather data and crop-specific parameters for potatoes and onions. The calendars revealed that water depth varied, depending on climate, crop type and growth stage. / Thesis (PhD)--University of Pretoria, 2009. / Plant Production and Soil Science / unrestricted