Cowpea yields in Mozambique can be increased through breeding farmers’ accepted cultivars with drought tolerance and stability across environments. A study was conducted in the southern region of Mozambique to: (1) determine farmers perceptions on major constraints limiting cowpea production and identify preferences regarding cultivars and traits, (2) determine the variability of selected cowpea germplasm for drought tolerance, (3) determine the gene action controlling drought tolerance, yield and yield components in cowpea, and (4) assess the genotype × environment interaction and yield stability of cowpea genotypes under drought-stressed and non-stressed conditions. The study on farmers’ perceptions about the major constraints limiting cowpea production and preferences regarding cowpea cultivars and traits established that cowpea was an important crop, cultivated for its grain, leaves and fresh pods for household consumption and the market. The study revealed that cowpea grain and leaves were equally important across the three districts in the study. Differences in accessibility to markets between districts influenced the ranking of grain and leaves among districts. Grain was more important in Bilene and Chibuto districts which are situated far from the major urban centre, Maputo, while leaves were more important in Boane district which is near the major market of Maputo. Fresh pods were important in Bilene district which is situated along the major highway connecting Maputo and other provinces. Drought was the most important production constraint followed by aphids, bruchids and viral diseases. The criteria used by farmers to select cowpea varieties included high grain and leaf yield, large seed size, earliness, smoothness of the testa and potential marketability of the variety. The implication of this study is that different types of varieties need to be developed for different areas. Dual-purpose or grain-type varieties need to be developed for areas situated far away from the major markets while varieties for leaf production need to be bred for areas near major markets. During the breeding process, a selection index needs to be adopted whereby drought tolerance, high grain and leaf yield, large seed size, smooth testa, earliness, aphids and bruchids resistance should be integrated as components of the index. High grain yield should receive high weight for varieties developed for areas located far from major markets while high leaf yield would receive high weight for varieties developed for areas located near major markets. The study on variability of cowpea germplasm collections for drought tolerance revealed wide genotypic variability among the tested germplasm. Biplot displays indicated that the genotypes could be grouped into four categories according to their drought tolerance and yielding ability as indicated below: high yielding-drought tolerant (group A), high yielding-drought susceptible (group B), low yielding-drought tolerant (group C), and low yielding-drought susceptible (group D). Examples of high yielding-drought tolerant genotypes were Sh-50, UC-524B, INIA-24, INIA-120, IT96D-610 and Tete-2. Stress tolerance index was the best criterion for assessing genotypes for variability in drought tolerance because it enabled the identification of high yielding and drought tolerant genotypes (group A). The assessment on gene action controlling drought tolerance (stay-green), yield and components indicated that both additive and non-additive effects were involved in controlling all of these traits. Additive gene action was more important than non-additive gene affects in controlling stay-green, days to flowering, number of pods per plant, number of seeds per pod and hundred seed weight. Under no-stress conditions, additive gene action was more important than non-additive gene action while under drought-stressed conditions, non-additive gene effects were more important than additive gene effects. Stay-green can easily be assessed visually in early segregating populations while yield and yield related traits cannot. Hence, selection for drought tolerance using the stay-green trait would be effective in early segregating generations while selection for yield and number of pods per plant would be effective in late segregating generations. Selection for yield could be conducted directly under no-stress conditions and indirectly using the number of pods per plant under drought stress conditions. Genotype INIA-41 would be the most desirable to use as a parent for drought tolerance and IT93K-503-1 would be the most desirable to use as a parent for drought tolerance and yield. The assessment on genotype × environment interaction and cowpea grain yield stability for forty-eight (48) cowpea genotypes grown under drought-stressed and non-stressed conditions indicated that cross-over genotype × environment interactions were present for yield indicating that genotypes responded differently to varying environmental conditions. Genotypes adapted to specific environmental conditions could be identified. Genotypes IT-18, INIA-51, INIA-51A and Nhavanca were adapted to non-stressed environments that were either drought stressed or non-stressed while VAR-11D was adapted to low yielding, stressful environments. Genotypes INIA-23A, INIA-81D, INIA-24, INIA-25, INIA-16 and INIA-76 were high yielding and stable while genotypes IT-18, INIA-51, INIA-51A, Nhavanca and VAR-11D were high yielding and unstable. Genotypes Bambey-21, INIA-36, INIA-12 and Monteiro were consistently low yielding and stable except INIA-12 that was consistently unstable. Chókwè was a high yielding environment and suitable for identifying high yielding genotypes but not ideal for selection because it was not representative of an average environment while Umbeluzi was low yielding and not ideal for selection. Overall, the study revealed that genetic improvement of drought tolerance and yield would be feasible. Potential parents for genetic improvement for yield and drought tolerance were identified. However, further studies for assessing yield stability of cowpea genotypes are necessary and could be achieved by including more seasons and sites to get a better understanding of the genotype × environment interaction and yield stability of cowpea in Mozambique. / Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2010.

Sorghum [Sorghum bicolor (L.) Moench] is one of the most important cereal crops grown in arid and semi-arid regions of the world. The North Eastern regions of Ethiopia are known for its high sorghum production and genetic diversity, and proneness to moisture stress. Globally, moisture stress is one of the major sorghum production constraints limiting genetic gain through breeding. Although, the importance of Ethiopia’s sorghum germpalsm has been widely recognized both nationally and internationally, the genetic potential of the germplasm has not yet been fully assessed and exploited in breeding programmes. Therefore, the objectives of this study were: (1) to evaluate sorghum production systems and patterns, major production constraints and cropping mechanisms, varietal diversification, farmers’ criteria for choosing varieties over time and space, and adoption of improved varieties, (2) to assess the agro-morphological and molecular diversity and population structure of lowland sorghum landraces collected from different geographic origins using morphological and SSR markers, (3) to assess the performance of sorghum landraces under moisture stress conditions and identify promising lines, and (4) to determine heterosis and combining ability of lowland sorghum landraces for grain yield, yield components and drought tolerance and to identify suitable parents for future hybrid cultivar development for the North Eastern Ethiopia or similar environments. A survey was conducted in the North Eastern Ethiopia sampling three Administrative Zones, six Districts and 12 Peasant Associations. Data was gathered from a total of 171 farmers and analyzed using SPSS statistical package. The results suggest that the performance of sorghum was generally poor mainly due to moisture stress, pests, diseases, weeds, farmland fragmentation due to demographic pressure, poor soil fertility, and poor performance of the local varieties. The productivity of sorghum was also largely hindered by the use of inherently poor yielding local landraces as farmers were forced to abandon their high yielding, big-headed and late maturing sorghum varieties due to the prevalence of recurrent moisture stress. The survey found that the region is as a rich source of genetic diversity and more than 70 distinct sorghum landraces were identified. The majority of the farmers grew their local landraces, despite the accessibility and availability of many improved sorghum varieties and production packages. Farmers were willing to adopt the improved varieties if they had farmers preferred attributes such as as red seed colour, tall with high biomass yield. To benefit the most from the available improved technologies, farmers have to be part of the breeding process right from the very beginning. Lowland sorghum accessions which exhibited farmer-desired traits were selected from the entire landrace germplasm collection at the national gene bank of Ethiopia. Field evaluations of the selected 278 landraces together with checks were held at Sirinka and Kobo agricultural research stations for 12 qualitative and 10 quantitative traits under stress and non-stress conditions. Two hundrad landraces were selected on the basis of their morphological distinctiveness and drought tolerance, in terms of earliness and yield stability. Molecular level diversity assessment was conducted using 30 SSR markers. Considerable magnitude of variation was observed among landraces between and within geographic origin for most of the traits studied. The morphological variability was also complemented by high molecular markers diversity. Thirty two pure lines were selected for inclusion as parents in the sorghum breeding programme for yield and drought tolerance. The selected lines were then crossed to four cytoplasmic male-sterile lines that had different cytoplasm systems (A1 to A4) using a line x tester mating design scheme. The 32 parents, together with the 128 hybrids and 4 check varieties were evaluated for grain yield, yield components traits and drought tolerance under stress and non-stress environments. Data were analysed using GenStat statistical package following a fixed effects model. Non-additive gene action was predominant in controlling plant height, grain yield, above ground biomass, grain filling duration, 100-seed weight and panicle weight, whereas additive gene action was found more important in controlling days to 50% anthesis and panicle length. Novel landraces with high GCA effects were selected including 214838-A, 242039-B, 75454, 73056-B, and 242050-A which will serve as potential parents for cultivar development. Similarly, the study identified new experimental hybrids i.e. ICSA 749 x 242039-B, ICSA 756 x 242049-B, ICSA 756 x 75454, ICSA 756 x 73059 and ICSA 756 x 214855 with high SCA effects and heterosis for grain yield which will be forwarded for further stability analysis and farmers participatory selections at representative growing environments. In general, the study identified invaluable sorghum germplasm and candidate hybrids useful for further breeding and conservation strategies. / Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2012.

This study aimed to characterize the physiological responses to drought of two sugarcane (Saccharum spp.) varieties in order to contribute information to study the physiological processes involved in plant adaptation to drought. To this end, an experiment was installed in a completely randomized design in a greenhouse at the Laboratory of Plant Physiology of the Center for Agrarian Sciences, Federal University of Alagoas, located in Rio Largo - AL (9° 28 S, 35° 49 W, with 127 m of altitude), and where the varieties RB855536 and RB867515 with contrasting responses to drought, were subjected to two, four, six and eight days of water stress and rehydration. The physiological and biochemical variables were analyzed, leaf water potential (w), solute potential (s), chlorophyll fluorescence (Fv/Fm and ФPSII), soluble sugars (glucose, fructose, sucrose) and starch and antioxidant enzymes, catalase (CAT), superoxide dismutase (SOD) and ascorbate peroxidase (APX). Water stress resulted in significant reduction in the w and s in two varieties studied, which did not support a drought of more than six days. After rehydration, the varieties studied, showed the ability to restore in the w and s. The RB867515 had greater reductions in the w and s in relation to RB855536. Both varieties studied presented chronic and dynamic photoinhibition gradually with the increase in time of drought stress and a strong reduction in the quantum potential efficiency (Fv/Fm) and effective (ΦPSII). The variety RB867515 showed higher capacity of recovery of the ΦPSII and rapair the effects of photoinhibition. The drought caused the decrease in the levels of sucrose and starch in two varieties, with greater reduction of recovery sucrose and starch compared to control, presented by RB867515. Increases in the levels of glucose and fructose were also observed in both varieties under water restriction. Changes in levels of sugars, indicating that rising levels of glucose and fructose occurred through the degradation of sucrose and starch, likely to hold the osmotic adjustment. The variety RB867515 showed higher activity of CAT and APX, in relation to RB855536, and SOD activity increased with increasing time of drought. In this work, the variety RB867515 showed greater efficiency in the activation of mechanisms of tolerance to drought stress. / Conselho Nacional de Desenvolvimento Científico e Tecnológico / O presente estudo teve por objetivo, caracterizar fisiologicamente as respostas à seca de duas variedades de cana-de-açúcar (Saccharum spp.), afim de contribuir com informações para o estudo dos processos fisiológicos envolvidos na adaptação das plantas ao déficit hídrico. Para tanto, instalou-se um experimento em delineamento inteiramente ao acaso, em casa de vegetação no Laboratório de Fisiologia Vegetal do Centro de Ciências Agrárias da Universidade Federal de Alagoas, localizado no município de Rio Largo - AL (9° 28 S, 35° 49 W, a 127 m de altitude), onde as variedades RB855536 e RB867515 com respostas contrastantes à seca, foram submetidas a dois, quatro, seis e oito dias de estresse hídrico e posterior reidratação. As variáveis fisiológicas e bioquímicas analisadas foram, potencial hídrico foliar (w), potencial de solutos (s), fluorescência da clorofila a (Fv/Fm e ФPSII), açúcares solúveis (glicose, frutose, sacarose) e amido, e atividade de enzimas antioxidantes, catalase (CAT), superóxido dismutase (SOD) e ascorbato peroxidase (APX). O estresse hídrico ocasionou em redução significativa no w e s nas duas variedades estudadas, as quais não suportaram um período de seca superior a seis dias. Após a reidratação, as variedades pesquisadas, mostraram capacidade de restabelecimento do w e s. A RB867515 teve maior decréscimo no w e s em relação a RB855536. Ambas as variedades pesquisadas apresentaram fotoinibição crônica e dinâmica gradativa com o aumento do tempo de estresse hídrico além de forte redução na eficiência quântica potencial (Fv/Fm) e efetiva (ΦPSII). A variedade RB867515 mostrou maior capacidade de recuperação do ΦPSII e de reparação aos efeitos da fotoinibição. O déficit hídrico causou a diminuição dos teores de sacarose e amido nas duas variedades, com maior nível de redução de sacarose e amido em relação ao controle, apresentado pela RB867515. Incrementos nos níveis de glicose e frutose também foram observados nas duas variedades sob restrição hídrica. As mudanças nos níveis de açúcares, indicam que a elevação nos níveis de glicose e frutose se deu através da degradação de sacarose e amido, provavelmente, no sentido de realizar o ajuste osmótico. A variedade RB867515 mostrou maior atividade de CAT e APX, em relação a RB855536, além de atividade crescente da SOD com aumento do tempo de seca. No presente trabalho, a variedade RB867515 revelou maior eficiência na ativação de mecanismos de tolerância ao estresse hídrico.

The soil water deficit is a problem that affects much of the cultivated areas in the world, especially those located in semi-arid regions. The sugarcane is a crop growing in northeastern Brazil, where the limitation of water availability affects growth and development of the plant, causing social and economic damage. Thus, this study aimed to evaluate, in a comparative way, the physiological aspects of six genotypes of cane sugarcane (RB931011, RB931013, RB72010, CL001, CL002 and CL003) submitted to water deficit during early vegetative growth under greenhouse conditions, at the Centre for Agrarian Sciences of the Federal University of Alagoas, Rio Largo, State of Alagoas, Brazil. The experimental design was completely randomized in factorial arrangement 6x3 with four replications. Water levels were defined as follows: control (80-100% of available water in soil - AW), mild stress (AW 40-60%) and severe stress (0-20% AW). Plants of 19 days were transplanted to pots. The treatments were applied to 80 days after transplanting and the pots were weighed daily to control of the humidity. Were taken periodically the SPAD readings, chlorophyll fluorescence parameters (quantum efficiency potential - Fv / Fm and effective quantum efficiency - ΦPSII) and measurements of leaf area. At the end of the experiment were measured the leaf water potential, plant height, tiller number and dry weight of leaf, stem plus sheath and shoot. Were also quantified the levels of photosynthetic pigments, osmoregulators (soluble sugars, proline, N-amino and proteins) and the enzyme activity of oxidative defense. Data were subjected to analysis of variance and average test (Tukey, 5% probability). Overall, there was reduction in the levels of photosynthetic pigments according to the intensity of stress. The w was reduced by the drought stress, however, the cultivar RB931011 showed no significant reduction in the water levels in the predawn and at noon, showed the smallest reduction when subjected to moderate stress. The Fv/Fm ratio was reduced by the treatments for genotypes RB931011, RB72910 and CL002, at noon, with high recovery rate during the night time (readings at 4:00 am). The RB72910 showed the greatest reduction in ΦPSII, SPAD index, content of chlorophyll b, total chlorophyll and ratio chla/chlb, but kept the carotenoids content. Regarding the osmoregulators, were increased the N-α-amino-soluble, free proline, soluble carbohydrate and were decreased the soluble protein amount, depending on water levels, independent of the genotypes. There was an increase in the level of malonilaldehyde (MDA) in response to water stress, more intense in RB931011 and RB931013 varieties. For the ascorbate peroxidase activity, there was a greater increase in CL002, even showing stability for catalase. Water stress reduced the initial growth and total dry matter production, and induced the activation of the oxidative defenses of sugarcane plants. RB72910 and CL001 genotypes showed higher tolerance to water deficit. / Conselho Nacional de Desenvolvimento Científico e Tecnológico / A deficiência hídrica dos solos é um problema que afeta boa parte das áreas cultivadas no mundo, sobretudo aquelas situadas nas regiões semi-áridas. A cana-de-açúcar é uma cultura em expansão no Nordeste do Brasil, onde a limitação da disponibilidade hídrica afeta o crescimento e o desenvolvimento da planta, causando prejuízos sócio-econômicos. Assim, o presente trabalho teve como objetivo avaliar, de maneira comparativa, aspectos fisiológicos de seis genótipos de cana-de-açúcar (RB931011, RB931013, RB72910, CL001, CL002 e CL003) submetidos ao déficit hídrico, durante a fase inicial de crescimento vegetativo, sob condições de casa-de-vegetação. O experimento foi realizado no Centro de Ciências Agrárias da UFAL, em Rio Largo, Estado de Alagoas, disposto em delineamento inteiramente ao acaso, em arranjo fatorial 6x3, com quatro repetições. Os níveis de água foram assim definidos: controle (80-100% da água disponível no solo - AD); estresse moderado (40-60% AD); e estresse severo (0-20% AD) e controlados pela pesagem diária dos vasos. Plantas de 19 dias de idade foram transplantadas para vasos de 15 L, e aos 80 dias após o transplantio, iniciou-se o experimento. Periodicamente foram realizadas leituras SPAD e de parâmetros da fluorescência da clorofila (eficiência quântica potencial - Fv/Fm e eficiência quântica efetiva - ΦPSII) e medições da área foliar. Ao final do experimento foram realizadas medidas do potencial hídrico foliar (w), diâmetro do colmo, altura de plantas, número de perfilhos e massa seca de folhas, colmo mais bainha e parte aérea. Também foram quantificados os teores de pigmentos fotossintéticos, osmorreguladores (açúcares solúveis, prolina, N-amino e proteínas) e atividade das enzimas de defesa oxidativa. Os dados foram submetidos à análise de variância e teste de média (Tukey a 5% de probabilidade). De modo geral, houve redução nos teores de pigmentos fotossintéticos de acordo com a intensidade do estresse. O w foi reduzido pelo déficit hídrico, entretanto, o genótipo RB931011 não apresentou redução significativa entre os níveis de água na ante-manhã e, ao meio dia, foi o que apresentou menor redução quando submetido ao estresse moderado. A relação Fv/Fm foi reduzida em função da deficiência hídrica para os genótipos RB931011, RB72910 e CL002, ao meio dia, apresentando alta taxa de recuperação durante o período noturno (leituras as 4:00h da manhã). O RB72910 apresentou a maior redução em ΦPSII, no índice SPAD e nos teores de clorofila a e b, clorofila total e relação chl a/chl b, porém manteve o teor de carotenóides. Em relação aos osmorreguladores, houve aumento para N-α-amino solúveis, prolina livre, carboidratos solúveis e redução para o teor de proteína solúvel, em função dos níveis de água, independente dos genótipos. Houve aumento no teor de aldeído malônico (MDA) em resposta ao estresse hídrico, sendo mais intenso nas variedades RB931011 e RB931013. Para a atividade da ascorbato peroxidase (APX), houve maior aumento no CL002, mesmo apresentando estabilidade para a catalase. O estresse hídrico reduziu o crescimento inicial e a produção de matéria seca total, einduziu a ativação das defesas oxidativas das plantas de cana-de-açúcar. A variedade RB72910 e o CL001 apresentaram-se mais tolerantes ao déficit hídrico.

Made available in DSpace on 2015-03-26T13:36:34Z (GMT). No. of bitstreams: 1 texto completo.pdf: 218982 bytes, checksum: 38c14fe52f8aa57350cdc239a6dbf920 (MD5) Previous issue date: 2006-02-16 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / This study aimed to identify potential photoprotective mechanisms and how they could be spatially and diurnally adjusted. It was conducted in an arabica coffee tree plantation (northsouth-oriented hedgerows) in Viçosa, Southeastern Brazil. Gas exchange, chlorophyll a fluorescence, and antioxidant system were examined throughout. Evaluations were performed in different canopy positions in August 2005 (dry, cold season with high insolation) when the net photosynthetic rate (A) of coffee tree is very low. This low A largely resulted from nonstomatal limitations. This, associated with an elevated interception of solar radiation, led to a discrete chronic photoinhibition of photosynthesis at noon, particularly in west-faced leaves. In a general way, decreased quantum yield of photosystem II (FPSII) paralleled an increased non-photochemical quenching, indicating a rise in heat dissipation. Apparent electron transport rate also rose during afternoon in west-faced leaves, that was largely a consequence from increased irradiance interception by those leaves, although decreases in both FPSII and photochemical quenching coefficient were observed. As a result, a higher excitation pressure should have occurred in west-faced leaves, thus leading to an increased fraction of irradiance neither used in photochemistry nor dissipated thermally. Regardless of their positions, coffee leaves showed similar ability for energy dissipation into photochemical and nonphotochemical processes. This suggests that the plant might dissipate satisfactorily the excess of absorbed light, i.e. the coffee tree might plastically adjust its photosynthetic machinery to changing irradiance. Steeper leaf angles were related to greater irradiance interception in the upper stratum leaves, particularly in the west side. Chlorophyll and carotenoid concentrations were higher in lower stratum leaves. There was no change in chlorophyll a/b ratio, regardless of the treatments. Activities of superoxide dismutase (SOD), catalase (CAT), and ascorbate peroxidase (APX) generally were similar among different sampled leaves. However, a greater SOD activity was found in upper stratum leaves that could be associated with a larger hydrogen peroxide concentration in those leaves. Little, if any, differences in the activities of SOD, CAT, APX and glutathione peroxidase were observed in paraquat-treated leaves. A stronger decrease in the maximum efficiency of photosystem II was registered in paraquattreated west-faced leaves, similarly to what occurred in the field. Despite west-faced leaves being naturally submitted to a high-light stress, malondialdehyde accumulated in a similar way as in the leaves from other canopy positions. Since the activity of antioxidant enzymes was similar irrespective of leaf position, some other photoprotecting systems should also be of utmost importance for protection of the photosynthetic machinery, particularly in west-faced leaves when exposed to bright irradiance. / O comportamento das trocas gasosas, dos parâmetros de fluorescência da clorofila a e do sistema antioxidativo foi estudado em plantas adultas de café arábica cultivadas em campo e orientadas no sentido norte-sul, em Viçosa-MG, objetivando-se identificar potenciais mecanismos de fotoproteção e avaliar como tais mecanismos se ajustariam espacial e diurnamente. Para isso, procederam-se às avaliações em diferentes posições da copa, em agosto de 2005, época relativamente fria, seca e com alta insolação, na qual o cafeeiro exibe taxas muito baixas de fotossíntese. As baixas taxas fotossintéticas foram largamente resultantes de limitações não-estomáticas. Isso, aliado à elevada interceptação da radiação solar, particularmente pelas folhas da face oeste à tarde, traduziu-se numa discreta fotoinibição crônica da fotossíntese. De modo geral, menores valores do rendimento quântico fotoquímico do fotossistema II (FFSII) foram acompanhados por incrementos paralelos no coeficiente de extinção não-fotoquímica, indicando aumento na capacidade de dissipação de calor. Ademais, a taxa de transporte aparente de elétrons aumentou, à tarde, nas folhas da face oeste, fato largamente associado à elevada irradiância interceptada por aquelas folhas, a despeito dos decréscimos observados em FFSII e no coeficiente de extinção fotoquímica. Como conseqüência, uma maior pressão de excitação ocorreu nas folhas da face oeste, resultando no aumento da proporção de energia não utilizada na fase fotoquímica e nem dissipada termicamente. De modo geral, as folhas do cafeeiro, independentemente de faces ou estratos, apresentaram capacidade similar de dissipação da radiação fotossinteticamente ativa em processos fotoquímicos e não-fotoquímicos. Isso sugere que o cafeeiro pode dissipar, de forma aparentemente satisfatória, o excesso da energia absorvida, apresentando capacidade de resposta plástica de sua maquinaria fotossintética às variações da irradiância. Maiores ângulos foliares mostraram-se relacionados à maior interceptação de luz pelas folhas dos estratos superiores, principalmente na face oeste da copa. A concentração de clorofilas e carotenóides foi maior em folhas dos estratos inferiores, mas não houve nenhuma alteração na razão clorofila a/clorofila b. As atividades da dismutase do superóxido (SOD), da catalase (CAT) e da peroxidase do ascorbato (APX), foram, de modo geral, semelhantes nas folhas, independentemente de estratos e faces. Somente uma maior atividade de SOD em folhas dos estratos superiores foi observada, fato que pode estar associado a maior acúmulo de peróxido de hidrogênio nas folhas daqueles estratos, independentemente de faces. Poucas foram as diferenças nas atividades da SOD, da CAT, da APX e da peroxidase da glutationa entre folhas das diferentes faces e estratos, quando se induziu o estresse oxidativo com Paraquat, mas uma maior redução da eficiência fotoquímica nas folhas dos estratos da face oeste foi observada, à semelhança do que ocorreu em condições de campo. Apesar de as folhas localizadas na face oeste da copa estarem submetidas a um maior estresse luminoso e, portanto, sujeitas de forma mais marcante ao estresse oxidativo, não houve danos celulares consideráveis, estimados pelo acúmulo de aldeído malônico. Uma vez que a atividade das enzimas estudadas foi muito similar, independentemente da posição na copa, sugere-se que outros sistemas de fotoproteção possam ter maior importância na proteção da maquinaria fotossintética, particularmente nas folhas da face oeste quando expostas à elevada irradiância.

Conselho Nacional de Desenvolvimento CientÃfico e TecnolÃgico / FundaÃÃo Cearense de Apoio ao Desenvolvimento Cientifico e TecnolÃgico / As membranas biolÃgicas sÃo alvos preferenciais dos efeitos deletÃrios do estresse hÃdrico, induzidos por aÃÃo de enzimas hidrolÃticas e espÃcies reativas do oxigÃnio (ERO), ambas estimuladas durante o estresse. A biossÃntese dos lipÃdeos dos cloroplastos pode ser importante para a tolerÃncia ao estresse hÃdrico e para a recuperaÃÃo apÃs reidrataÃÃo. Nesse trabalho nos estudamos o metabolismo dos cloroplastos, monogalactosil-diacilglicerol (MGDG), digalactosil-diacilglicerol (DGDG), sulfoquinovosil-diacilglicerol (SQDG), phosphatidil-glicerol (PG), no Ãmbito do dÃficit hÃdrico e da reidrataÃÃo apÃs o fim do estresse. Com este intuito, nos medimos o teor dos lipÃdeos da folhas, acompanhamos a incorporaÃÃo do precursor 14C-acetato nos lipÃdeos e analisamos a expressÃo dos genes codificadores das enzimas de sÃntese chave dos lipÃdeos (MGD1, MGD2, DGD1, DGD2, SQD2 e PGP1) durante o estresse hÃdrico e apÃs a reidrataÃÃo. Visando de uma melhor compreensÃo da relaÃÃo entre o metabolismo destes lipÃdeos e a tolerÃncia a seca, nos trabalhamos com duas cultivares de Vigna unguiculata L. Walp, uma tolerante (cv. EPACE) e outra sensÃvel (cv. 1183) a seca. Por meio de varredura diferencial de um biblioteca de cDNA de V.unguiculata, foram obtidas as seqÃÃncias completas dos cDNA dos genes VuMGD1, VuMGD2, VuDGD1, VuDGD2, VuSQD2 e VuPGP1. Os resultados mostram que em condiÃÃes de estresse hÃdrico a cultivar tolerante, alÃm de preservar seu teor de lipÃdeos durante a seca, à igualmente capaz de estimular a biossÃntese do DGDG aumentando significativamente a relaÃÃo DGDG:MGDG de suas membranas. NÃs sugerimos que o DGDG acumulado em condiÃÃo de seca à transportado para as membranas externas ao cloroplasto e que isso contribui para a tolerÃncia à seca. Os efeitos da desidrataÃÃo celular sobre as membranas tÃm conseqÃÃncias diretas sobre a capacidade das plantas a se recuperarem apÃs reidrataÃÃo. 48 horas apÃs a rega, a cv. sensÃvel 1183 nÃo à capaz de se recuperar em termos de teor de galactolipÃdeos e incorporaÃÃo do precursor. Na cv. tolerante, no entanto, o teor de DGDG permanece elevado, mesmo apÃs a reidrataÃÃo. Em conclusÃo, nossos resultados sugerem a importÃncia dos lipÃdeos membranares na tolerÃncia/sensibilidade das plantas ao dÃficit hÃdrico, em particular o balanÃo entre as classes lipÃdicas de propriedades fÃsico-quÃmicas diferentes (SQDG versus PG e DGDG versus MGDG) que poderiam afetar a estrutura e o funcionamento das membranas. / Membranes are main targets of degradation by reactive oxygen species and hydrolytic activities induced by drought. Chloroplasts lipid biosynthesis, especially galactolipids monogalactosyl-diacylglycerol (MGDG) and digalactosyl-diacylglycerol (DGDG) are important for plant tolerance to water deficit and for recovery after rehydration. In this thesis, we studied the metabolism of the chloroplast membrane lipids, MGDG, DGDG, sulphoquinovosyl-diacylglycerol (SQDG), phosphatidyl-glycerol (PG) under drought and during recovery from drought. Aiming this, we measured leaf lipids content, followed 14C-acÃtate incorporation and expression of genes coding for chloroplast membrane lipid synthases (MGD1, MGD2, DGD1, DGD2, SQD2 and PGP1) during drought and recovery. In order to better understand the relationship between drought tolerance and lipid metabolism, two cultivars of Vigna unguiculata L. Walp, one drought tolerant (cv. EPACE) the other drought susceptible (cv. 1183) were compared. The cDNA complete sequences for VuMGD1, VuMGD2, VuDGD1, VuDGD2, VuSQD2 and VuPGP1 were obtained from screening of a V.unguiculata cDNA library. The results showed that under water stress conditions, the tolerant cultivar, besides its ability to preserve its lipids pool despites drought, is able to strongly stimulate the DGDG biosynthesis, increasing the DGDG:MGDG ratio in its membranes. We suggest that DGDG accumulated under drought condition, when phosphate is deficient, is exported for extrachloroplastic membranes, and thus contributes to plant drought tolerance. Effects of loss of water on cell membranes have direct consequences on plant capacity to recover from stress. 48 hours after rewatering, the susceptible cv. 1183 was not able to fully recover from a moderate stress in terms of leaf galactolipid content and acetate incorporation into MGDG. In EPACE-1, MGDG leaf content remained unchanged after rehydration and DGDG remained higher than in the control plants. In conclusion, our results highlight the importance of membrane lipids in plant adaptation to water deficit and in their capacity to recover from stress. Of particular importance is the balance between lipid classes with various physico-chemical properties (SQDG versus PG, DGDG versus MGDG), since they most likely have a profound influence on membrane structure and function.

Submitted by Natalia de Souza Gonçalves (natalia.goncalves@ufpe.br) on 2016-10-10T12:47:42Z No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Dissertação Lumena Feitosa Duda.pdf: 3810732 bytes, checksum: 139d9ec8d60cf833b8f3dc69f96cd690 (MD5) / Made available in DSpace on 2016-10-10T12:47:42Z (GMT). No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Dissertação Lumena Feitosa Duda.pdf: 3810732 bytes, checksum: 139d9ec8d60cf833b8f3dc69f96cd690 (MD5) Previous issue date: 2014-02-19 / Capes / O aumento na demanda por produção de cana-de-açúcar exige maior investimento na geração de novas variedades tolerantes aos estresses bióticos e abióticos, melhorando as condições de plantio, colheita e produtividade. A produção de variedades tolerantes à seca, exige o conhecimento detalhado dos aspectos morfofisiológicos que distinguem entre os indivíduos mais vantajosos para o plantio. Dessa forma, esse trabalho teve como objetivo avaliar alguns aspectos anatômicos, espessura do mesofilo, arquitetura estomática, caracterização epidérmica, análises de trocas gasosas e potencial hídrico foliar entre as variedades sensíveis RB855536 e RB72454 e as tolerantes RB867515 e RB92579 ao déficit hídrico. Verificou-se que o potencial hídrico foliar (w) foi menor na estação seca, em especial àquele medido ao meio dia. A variedade RB72454 obteve menor valor de w (-2,21 MPa). Durante a estação seca, ocorreu uma compactação do mesofilo em cerca de 30% para todas as variedades. A taxa de assimilação líquida de CO2 (AL) apresentou maiores reduções na estação seca, para todas as variedades, com menores médias na variedade RB92579. Por outro lado, a condutância estomática (gs) foi maior na variedade RB855536. A morfologia dos estômatos e sua distribuição nas superfícies epidérmicas diferiu de acordo com a estação, com menor densidade estomática na estação seca em ambas faces. Uma ampla deposição de cera epicuticular sobre o poro estomático parece ser uma das principais características que determinam a menor gs no período seco. / The increase in demand for production of sugar cane requires greater investment in new generation tolerant varieties to biotic and abiotic stresses, improving conditions for planting, harvesting and productivity. The production of drought tolerant varieties requires a detailed knowledge of Morphophysiological aspects that distinguish between the most viable individuals for planting. This study aimed to evaluate some anatomical aspects, mesophyll thickness, stomatal architecture, epidermal characterization, analysis of gas exchange and leaf water potential between susceptible varieties RB855536 and RB72454 and RB92579 and RB867515 drought tolerant. It was found that the leaf water potential (w) was lower in the dry season, especially to that measured at noon. The variety RB72454 got smaller value of w (-2.21 MPa). During the dry season, there was a compression of the mesophyll by about 30% for all varieties. The rate of net CO2 assimilation (AL) showed greater reductions in dry season for all varieties with lower averages in variety RB92579. On the other hand, stomatal conductance (gs) was higher in the range RB855536. The morphology and distribution of stomata in epidermal surfaces differed according to season, with lower stomatal density in the dry season on both side. An extensive deposition of epicuticular wax on the stomatal pore seems to be one of the main characteristics that determine the lowest gas in the dry season.

A caracterização e o entendimento das variações genômicas e morfológicas, bem como a estrutura genética de variedades locais armazenadas em bancos de germoplasma é importante para sua efetiva utilização em programas de melhoramento visando tolerância a estresses. Neste trabalho um conjunto de 192 variedades oriundas de diferentes regiões geoclimáticas do Japão foram testadas quanto à suas características morfológicas e produtivas, utilizando ensaios de campo e metodogias de fenotipagem de alto desempenho. A fenotipagem por meio da metodologia de camadas de herbicida (Aminotriazol+Diuron+2,4D, 100mg/plant) alocado a 30 centímetros de profundidade foi possível detectar variação entre as variedades para comprimento de raiz e velocidade de emissão de raiz sendo possível a distinção de variedades com sistema radicular profundo e sistema radicular superficial baseando-se na sua pontuação no ensaio de herbicida, destacando-se 20 genótipos como possíveis doadores de genes para comprimento, densidade e velocidade de emissão de raízes. Ensaios a campo foram conduzidos em 4 localidades expondo as variedades as mais distintas condições climáticas, buscando analisar a diversidade fenotípica para caracteres agromorfológicos. Os dados fenotípicos obtidos pelos marcadores morfológicos geraram um total de 15 grupos de acessos quando utilizados os 13 caracteres avaliados. A média do índice de sensibilidade a seca foi de 0,99 havendo materiais tolerantes com índices próximos a 0,6 e materiais sensíveis com índice próximos a 1,12. Os 384 marcadores SNP detectaram um total de 73728 alelos indicando alta porcentagem de A (40,8%) e G (34,6%) comparado com C (15,6%) e T (3,6%). Quanto aos heterozigotos, a maior porcentagem foi observada de A/G (0,54%) e a menor porcentagem de A/T (0,04%), sendo a maior parte dos heterozigotos observados nos cromossomos 3 e 8 comparado com outros cromossomos. As análises caracterizaram os acessos japoneses como 98,4% pertencentes à subespécie Japônica. Para associação entre marcadores e fenótipos, foi utilizada a abordagem de modelo linear misto (MLM), o qual incorpora informações de estrutura populacional e parentesco. Os resultados obtidos deverão ser investigados futuramente a fim de confirmar as associações em diferentes populações. O aumento do estresse hídrico teve efeito significativo no desempenho dos acessos, e a interação genótipo e estresse hídrico foi significativa para rendimento final e tamanho de panícula. Entre os acessos estudados foi observada variação genética para as características relacionadas com a tolerância a estresse hídrico e encontraram-se acessos com reduções no rendimento devido ao déficit hídrico comparáveis com o das testemunhas, embora com tamanho de panícula menor, inclusive em condições ótimas. Dos 384 marcadores utilizados, 10 foram responsáveis por associações significativas com o índice de sensibilidade a seca, com base nos diferentes métodos de correção para múltiplos testes. Estas associações foram selecionadas para verificar o efeito alélico sobre o genótipo observado, gerando informações preliminares para a aplicação futura de seleção assistida por marcadores (SAM). O tamanho dos blocos de ligação foram estimados em ~100 kb (r < 0,05) e ~75kb (r < 0,1). / Germplasm characterization and the knowledge of its diversity and population structure are important to effective utilization of genetic resources in breeding programs specially drought breeding program. In this work 192 landraces from all over Japan were evaluate for their morphological and productive characteristics, using field trials and high throughput screening methods. The screening using herbicide barrier approach at 30 cm depth was able to detect genetic diversity between the landraces for root length and clearly distinguish between deep root landraces and shallow root landraces. With this approach was possible to select 20 landraces with deep root system as possible donor of drought tolerance genes. Aiming characterize the landraces to agromorphological characteristics field trials were carried in 4 different locations exposing the landraces for a diversity environment effect. Using 13 traits phenotypically data generate a total of 15 groups during cluster analysis. The average result for drought sensitive index was 0.99, however this results presents a huge variability having landraces with scores about 0.6 to landraces with score up to 1.12. Drought effect was huge and statistically significant affecting directly yield and panicle size. The landraces presented genetic variability for drought tolerance and some landraces presenting yield and panicle size reductions due to drought comparable with drought-tolerant controls were detected. A total of 73728 alleles were detected by the SNP markers, indicated a high percentage of A (40,8%) and G (34,6%) alleles compared to C(15,6%) and T (3.6%). Heterozygocity of A/G was highest (0,54%) and lowest in A/T (0.04%). Of 3 chromosomes of rice, chromosome 8 produced highest percentage of heterozygocity compared to other chromosomes. Accessions were classified as 98.4% belonging to japônica subspecies. Association between markers and phenotypes was performed using a mixed linear approach (MLM), which incorporates information regarding population structure and kinship. Among the 384 markers used, 10 were responsible for significant associations with drought sensibility index, based on different criteria to correct for multiple tests.These associations were selected to determine the allelic effects over the traits, in order to generate preliminary data for marker assisted selections (MAS). Estimated size of haplotype blocks were ~100 kb (r <0.05) and ~75 kb (r <0.1). Future studies should confirm marker trait associations here found using different populations.

Orientador: Marcelo Menossi Teixeira, Andrea Akemi Hoshino / Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-21T05:45:50Z (GMT). No. of bitstreams: 1 Souza_CesarBuenode_D.pdf: 2755272 bytes, checksum: 945d7a45c10893c9a58603f1e04813ef (MD5) Previous issue date: 2012 / Resumo: O Brasil é o maior produtor mundial de cana-de-açúcar e no cenário atual, em que o aumento na busca por energia renovável é visível, o aumento na produtividade da cana é de extremo interesse para o setor sucroalcooleiro. Estresses abióticos influenciam grandemente a produtividade de espécies como a cana e, sendo assim, estudos relacionados com a tentativa de diminuir esse impacto na produtividade são de grande importância. A seca é o estresse ambiental que mais causa prejuízos ao agronegócio e por esse motivo é muito desejável que se desenvolvam novas variedades de cana-de-açúcar que sejam mais tolerantes a esses estresses e com isso o setor sucroalcooleiro será largamente beneficiado. A transgenia é uma das ferramentas utilizadas na produção de novos cultivares comerciais com características agronômicas interessantes e, para o seu sucesso, a identificação de genes com potencial para melhorar essas características faz-se necessária. Sendo assim, o maior objetivo desse trabalho foi a avaliação do potencial biotecnológico de seis genes de cana-de-açúcar que são modulados por seca. Os genes aqui estudados tiveram seu perfil de resposta a seca anteriormente avaliado por microarranjos de DNA e foram selecionados como candidatos para a produção de um novo cultivar de cana-de-açúcar que seja mais tolerante a estresses abióticos. Esses genes foram analisados em plantas transgênicas de tabaco, cana-de-açúcar e/ou Brachypodium. A superexpressão de dois genes de cana em tabaco conferiu maior tolerância a seca e salinidade das plantas transgênicas quando comparadas às selvagens e a proteção de seus usos na produção de plantas tolerantes a estresses abióticos foi solicitada. Há evidências de que um terceiro gene de cana conferira tolerância ao estresse oxidativo em plantas transgências de cana. Além disso, outros três genes de cana foram inseridos em Brachypodium, mas os eventos gerados ainda não foram avaliados. Com o trabalho desevolvido foi, portanto, possível gerar plantas transgênicas tolerantes a estresses abióticos e com isso é possível concluir que a seleção de genes candidatos para melhorias de características agronômicas de interesse através de microarranjos é algo que deve ser explorado e pode-se concluir, ainda, que alguns dos genes analisados estão envolvidos na resposta a seca e/ou aos estresses ambientais em geral / Abstract: Brasil is the largest producer of sugarcane in the world and the seeking for renewable energy is currently visible what makes the increase of sugarcane productivity highly desirable. Abiotic stresses greatly influence the productivity of species such sugarcane. Therefore, studies related to the reduction of these impacts on productivity are highly important. Drought is the environmental stress that causes more damage to agribusiness and because of it the development of new cultivars with higher tolerance to abiotic stresses is desirable since the sugar and ethanol sector will be largely benefited. Transgenic plants production is one of the tools that have been used in the development of new cultivars with interesting agronomic traits and for its success identifying genes that can improve these characteristics is necessary. Thus, the main objective of this study was evaluating the biotechnological potential of six drought-modulated genes from sugarcane. The genes studied here have their drought response profile previously showed by microarray and were selected as candidates for the production of a new sugarcane cultivar with higher tolerance to abiotic stresses. These genes were analyzed in transgenic tobacco, sugarcane and/or Brachypodium plants. The overexpression of two sugarcane genes in tobacco conferred higher drought and salinity tolerance in tobacco plants compared to wild-type and the protection of their uses in the production of plants with higher tolerance to abiotic stress was requested. There are evidences that a third sugarcane gene confers tolerance to oxidative stress in transgenic sugarcane. Furthermore, three other genes were inserted in Brachypodium but these transgenic events were not yet analyzed. With this work was possible to produce transgenic plants that are tolerant to abiotic stresses what let us to conclude that the selection of candidate genes to improve agronomic traits by microarrays is useful and that some of the analyzed genes are involved into drought and/or abiotic stresses in general responses / Doutorado / Genetica Vegetal e Melhoramento / Mestre em Genética e Biologia Molecular

Common bean is a major dietary component in several countries, but its productivity is negatively affected by abiotic stresses. Dissecting candidate genes involved in abiotic stress tolerance is a paramount step toward the improvement of common bean performance under such constraints. Thereby, this thesis presents a systematic analysis of the DEHYDRATION RESPONSIVE ELEMENT-BINDING (DREB) gene subfamily, which encompasses genes that regulate several processes during stress responses, but with limited information for common bean. First, a series of in silico analyses with sequences retrieved from the P. vulgaris genome on Phytozome supported the categorization of 54 putative PvDREB genes distributed within six phylogenetic subgroups (A-1 to A-6), along the 11 chromosomes. Second, we cloned four novel PvDREB genes and determined their inducibility-factors, including the dehydration-, salinity- and cold-inducible genes PvDREB1F and PvDREB5A, and the dehydration- and cold-inducible genes PvDREB2A and PvDREB6B. Afterwards, nucleotide polymorphisms were searched through Sanger sequencing along those genes, revealing a high number of single nucleotide polymorphisms within PvDREB6B by the comparison of Mesoamerican and Andean genotypes. The nomenclature of PvDREB6B is discussed in details. Furthermore, we used the BARCBean6K_3 SNP platform to identify and genotype the closest SNP to each one of the 54 PvDREB genes. We selected PvDREB6B for a broader study encompassing a collection of wild common bean accessions of Mesoamerican origin. The population structure of the wild beans was accessed using sequence polymorphisms of PvDREB6B. The genetic clusters were partially associated with variation in latitude, altitude, precipitation and temperature throughout the areas such beans are distributed. With an emphasis on drought stress, an adapted tube-screening method in greenhouse conditions enabled the phenotyping of several drought-related traits in the wild collection. Interestingly, our data revealed a correlation between root depth, plant height and biomass and the environmental data of the location of the accessions. Correlation was also observed between the population structure determined through PvDREB6B and the environmental data. An association study combining data from the SNP array and DREB polymorphisms enabled the detection of SNP associated with drought-related traits through a compressed mixed linear model (CMLM) analysis. This thesis highlighted important features of DREB genes in common bean, revealing candidates for further strategies aimed at improvement of abiotic stress tolerance, with emphasis on drought tolerance / O feijoeiro é um componente essencial na dieta em diversos países, no entanto, sua produção é afetada negativamente por estresses abióticos. O estudo de genes candidatos envolvidos na adaptação aos estresses é uma etapa fundamental para o melhoramento da performance do feijoeiro sob tais estresses. Desse modo, esta tese apresenta uma análise sistemática da subfamília de genes DEHYDRATION RESPONSIVE ELEMENT-BINDING (DREB), que reúne genes envolvidos em diversos processos em resposta a estresses, mas pouco estudados no feijoeiro. Primeiramente, uma série de análises in silico com sequências de feijoeiro obtidas da plataforma Phytozome possibilitaram a categorização de 54 genes PvDREB putativos, distribuídos em seis subgrupos (A-1 até A-6) nos 11 cromossomos da espécie. Posteriormente, quatro novos genes PvDREB foram clonados e seus padrões de inducibilidade foram determinados. PvDREB1F e PvDREB5A foram induzidos por desidratação, baixa temperatura e salinidade, enquanto PvDREB2A e PvDREB6B foram predominantemente induzidos por desidratação e baixa temperatura. Polimorfismos de nucleotídeos foram buscados através de sequenciamento por método derivado de Sanger, revelando elevado número de SNP no gene PvDREB6B. A nomenclatura desse gene foi discutida detalhadamente ao longo da tese. A plataforma de marcadores SNP BARCBean6K_3 foi acessada para identificar o SNP mais próximo de cada um dos 54 PvDREB. O gene PvDREB6B foi selecionado para um estudo mais amplo, envolvendo uma coleção de acessos selvagens de origem Mesoamericana. A estrutura populacional destes genótipos foi analisada a partir de polimorfismos na sequência de PvDREB6B. Os grupos genéticos apresentaram associação parcial com variação da latitude, altitude, precipitação e temperatura das áreas em que os acessos naturalmente ocorrem. Com ênfase no estudo do déficit hídrico, uma plataforma de fenotipagem destes acessos em casa de vegetação, utilizando um sistema de tubos plásticos, foi elaborada para a análise de diversos parâmetros relacionados ao estresse por déficit hídrico. Os dados revelaram correlação entre profundidade de raízes, altura das plantas e a biomassa e as variáveis ambientais de cada local. A correlação também foi detectada entre a estrutura populacional estudada por PvDREB6B e os dados ambientais. Finalmente, um estudo de associação genética foi realizado entre os SNP da plataforma e ligados a DREB e os parâmetros fenotípicos, permitindo a identificação de marcadores SNP associados a caracteres específicos, usando um modelo linear misto (CMLM). Esta tese apresentou importantes aspectos sobre os genes DREB em feijoeiro, revelando candidatos para seu uso em estratégias de melhoramento para tolerância a estresses abióticos, com ênfase em déficit hídrico