Abiotic transformation of compounds in the natural environment by metal oxides plays a significant a role in contaminant fate and behavior in soil. The ability of birnessite, ferrihydrite and green rust to abiotically transform insensitive munitions compounds (IMCs) parent (2,4 dinitroanisole [DNAN] and 3-nitro-1,2,4-triazol-5-one [NTO]), and daughter products (2-methoxy-5-nitro aniline [MENA], 2,4-diaminoanisole [DAAN]of DNAN; and 5-amino-1, 2, 4-triazol-3-one [ATO] of NTO) was studied in batch reactors under strictly controlled pH and ionic strength. The objectives of the study were to (i) assess the abiotic transformation potential of soluble DNAN, MENA, DAAN, NTO and ATO by birnessite, ferrihydrite and green rust, and (ii) identify inorganic reaction products. The study was carried out at metal oxide solid to IMC solution ratios (SSR) of 0.15, 1.5 and 15 g kg⁻¹ for birnessite and ferrihydrite and 10 g kg⁻¹ for green rust. Aqueous samples were collected at time intervals between 0 to 3 days after the reaction initiation and analyzed using HPLC with UV detection. Results indicated that DNAN was resistant to oxidation by birnessite and ferrihydrite at given solid to solution ratios. MENA was susceptible to rapid oxidation by birnessite (first order rate constant, 𝑘=1.36 h⁻¹ at 15 g kg⁻¹ SSR). The nitro groups from MENA largely mineralized to nitrite (NO₂⁻). In contrast, ferrihydrite did not oxidize MENA. DAAN was susceptible to oxidation by both birnessite and ferrihydrite, but about a six times higher oxidation rate was observed with birnessite (𝑘=1.18 h⁻¹) as compared to ferrihydrite (𝑘=0.22 h⁻¹) at an SSR of 1.5 g kg⁻¹. There was a complete loss of DAAN from solution after 5 min with birnessite at an SSR 15 g kg⁻¹ (𝑘≥90.5 h⁻¹). CO₂ evolution experiments indicate mineralization of 15 and 12 % of carbon associated with MENA and DAAN, respectively; under aerobic conditions with birnessite at an SSR of 15 g kg⁻¹. NTO was resistant to oxidation by birnessite and ferrihydrite at any SSR; however, there was slight initial loss from solution upon reaction with ferrihydrite at 0.15 and 1.5 g kg⁻¹ SSR and complete loss at 15 g kg⁻¹ SSR due to adsorption. ATO was susceptible to oxidation by birnessite and sorption by ferrihydrite. The first order rate constants (𝑘) for ATO with birnessite at 0.15 and 1.5 g kg⁻¹ SSR are 0.04 and 3.03 h⁻¹ respectively. There was complete loss of ATO from solution with birnessite at 15 g kg⁻¹ SSR (𝑘 ≥ 90.2 h⁻¹) within 5 min of reaction. Transformation products analysis revealed urea, CO₂ and N₂ as major reaction products with 44 % urea recovery and recovery of 51.5 % of ATO carbon as CO₂ and 47.8 % of ATO nitrogen as N₂ at 15 g kg⁻¹ SSR. The oxidation of ATO in the presence of birnessite was found to be independent of dissolved O₂. The results indicate that ATO, the major reductive (bio)transformation product of NTO, is readily oxidized by birnessite in soil. NTO was found strongly sorbed to ferrihydrite as compared to that of ATO. The results of the green rust experiment indicate rapid abiotic reduction of parent compounds NTO and DNAN to their reduced aminated daughter products. NTO was generally reductively transformed to 5-amino-1, 2, 4-triazol-3-one (ATO) within 10 min and completely reacted in 20 min. DNAN was rapidly transformed to its reduced daughter products MENA and 4-methoxy-5-nitroaniline (iMENA). The reduction occurred with a distinctive, staggered regioselectivity. Over the first 10 min, the para-nitro group of DNAN was selectively reduced, generating iMENA. Thereafter the ortho-nitro group was preferentially reduced, generating MENA. Both iMENA and MENA were subsequently transformed to the final reduction product DAAN within 1 day. X-ray absorption near edge spectroscopy data suggested oxidative transformation of green rust to lepidocrocite-like mineral forms, accounting for 94 % of the mineral products in the case of NTO reaction as compared to 62 % in the case of DNAN. The results taken as whole suggest that complete abiotic transformation of IMCs could be achieved by coupled stepwise green rust and birnessite treatments.

The requirements of true frogs on breeding sites were monitored at 19 locations in the Hradec Králové Region and the Central Bohemia Region and Vysočina Region. Specific sites were selected according to the high variability. Breeding sites are mutually different representation of true frogs, water volume, depth, water and surrounding vegetation. The data of oviposition three different species of czech frogs - moor frog (Rana arvalis), agile frog (Rana dalmatina) and common frog (Rana temporaria) - were collected in the spring months, from the end of March until the end of April, from 2012 to 2015. During this period were collected data from 927 egg clutches of these species. The data were obtained by non-invasive way - measuring of inividual clutches directly on the reproductive site. At each breeding site was also recorded the required parameters of the breeding pond and the surrounding environment (presence of true frogs, the water volume and depth of the pond, the presence and density of aquatic vegetation, surrounding vegetation, air and water temperature, pH value of water, etc.). The results presented in this work confirm that the three species of true frogs have, in some parameters, different standards to select of breeding sites. This clarification of species- specific claims is also...

Certaines pratiques agricoles telles que l’épandage de produits résiduaires organiques (PRO) ou l’irrigation des sols peuvent être à l’origine de l’introduction de bactéries pathogènes de l’Homme dans les sols. Le sol joue alors un rôle central dans la dissémination de ces bactéries pathogènes dans les différents compartiments de l’environnement et peut contaminer les matières premières végétales, les animaux d’élevage mais également les ressources en eau. Le risque sanitaire associé à la dissémination de ces bactéries pathogènes doit être évalué afin de limiter le développement de maladies infectieuses. Dans ce contexte, l’incidence et le potentiel de survie de bactéries pathogènes a été étudié dans un large ensemble de sols provenant du territoire français (Réseau de Mesure de la Qualité des Sols : RMQS). Ce travail a pour objectif général d’identifier les facteurs biotiques et abiotiques des sols qui déterminent l’incidence et la survie de d’un pathogène strict, Listeria monocytogenes, et d’un pathogène opportuniste, Enterococcus faecalis, dans ces matrices environnementales Pour ceci, 2 approches ont été adoptées : la première approche a visé à mettre au point des systèmes de détection de PCR quantitative spécifiques pour ces deux espèces bactériennes afin de détecter et quantifier ces 2 bactéries dans un ensemble de 1200 ADN de sols présentant des paramètres pédoclimatiques contrastés. L. monocytogenes n’est pas détectée dans les ADN de sols avec la méthode moléculaire développée, dont le seuil de détection est d’environ 104 bactéries par gramme de sol. La détection moléculaire d’E. faecalis a été biaisée par un problème de contamination des ADN extraits. Après extraction de nouveaux ADN sur 150 échantillons de sol, E. faecalis a été détecté en faible quantité dans 5 des 150 sols testés (4% des sols) à des concentrations d’environ 102 bactéries par gramme de sol. Dans une deuxième approche, la survie de L. monocytogenes et d’E. faecalis a été suivie dans un ensemble de 100 sols caractérisés par des paramètres pédoclimatiques variables. Des microcosmes de sols ont été inoculés et la survie bactérienne a été évaluée par des méthodes culturales classiques. L. monocytogenes survit à long terme dans un plus grand nombre de sols qu’E. faecalis. Les sols favorables à la survie à long terme sont majoritairement (44%) les mêmes pour les 2 bactéries. Les sols sableux présentant un pH acide sont défavorables à la survie des 2 bactéries. Cependant des différences sont observées quant aux paramètres physico-chimiques des sols qui déterminent la survie de ces 2 bactéries. Ainsi, la survie de L. monocytogenes est corrélée positivement au taux de saturation en cations basiques, à la capacité d’échange cationique et à la concentration en calcium pour le court terme et à la teneur en argile pour le long terme. En revanche, pour E. faecalis, une corrélation négative avec la teneur en calcium total est trouvée pour la survie à court terme alors que la survie à long terme est négativement corrélée à la concentration en phosphore assimilable. Un effet inhibiteur de la microflore des sols a été mis en évidence sur la survie de L. monocytogenes et E. faecalis : il est statistiquement significatif pour les sols présentant un pH supérieur à 7 pour les 2 pathogènes. Les résultats obtenus vont permettre de modéliser la survie de ces pathogènes dans les sols en fonction de paramètres pédo-climatiques et donc à terme de mieux gérer les épandages de PRO en fonction des types de sols pour limiter la persistance de pathogènes dans les sols. / Soil contamination by bacterial pathogens can occur through manure, sewage sludge spreading or irrigation using waste water treatment plants effluents. Agricultural soils may act as reservoirs for these pathogens, play a significant role in their dissemination, leading to the potential contamination of food and water resources. Health risk associated with the occurrence of pathogens in environmental matrices has to be thoroughly evaluated. In this context, the objectives of this work were: i) to determine the prevalence of two pathogenic bacterial species (Listeria monocytogenes and Enterococcus faecalis) in a large collection of French soils originated from a systematic soil survey of the territory, called RMQS (16x16 km grid), ii) to determine major biotic and abiotic parameters driving the survival of bacterial pathogens in soils. Two approaches were used to reach these objectives: i) the prevalence of the two pathogens was monitored in 1200 soils using specific molecular detection tools (real time PCR using TaqMan probe detection system) and ii) the survival of the 2 pathogens, inoculated in soil microcosms, was determined over a 84 days incubation period under laboratory conditions, on a subset of 100 soils from the RMQS survey, using classical microbiological methods. L. monocytogenes was not detected in the set of 1200 soils (with a detection limit estimated to be 104 bacteria per gram of soil) using the molecular detection method, while E. faecalis was detected in approximately 4% of the soils tested (on a smaller set of 150 RMQS soils). The two bacterial pathogens were able to survive in the majority of soil, although L. monocytogenes survived in a greater number of soils. Soils where both pathogens survived represented 44% of soils. The survival of both pathogens is strongly impaired in sandy acidic soils. However, textural and chemical parameters driving survival of the two pathogens differed: L. monocytogenes survival is increased in soils with higher BCSR (basic cation saturation ration) and CEC (cation exchange capacity). L. monocytogenes long-term survival is favored by higher soil clay content. E. faecalis survival is impaired in soils with high total Ca content (calcareous soils). Soil microflora inhibits survival of both pathogens especially in soil with alcaline pH’s. The results of our study will allow implementing survival models for these two pathogens. Such data is invaluable for a better and safer managment of soil manuring using various organic residues.

Hevea brasiliensis est cultivé pour le caoutchouc naturel contenu dans le latex. Une exploitation intensive combinée aux stress environnementaux affectent la production de latex. L'encoche sèche ou Tapping Panel Dryness (TPD) est déclenché par un désordre physiologique complexe au sein des laticifères. Il est responsable d'une perte annuelle de production de 10 à 40% en fonction de l'âge de la plantation et des clones d'hévéa utilisés Le stress oxydatif, point de départ de la maladie, affecte l'écoulement, par la coagulation in situ des particules du caoutchouc. Chez les plantes, la réponse adaptative aux stress abiotiques dépend de la finesse de la régulation de l'expression des gènes. Ce contrôle se fait au niveau transcriptionnel mais également au niveau post-transcriptionnel. Les micro-ARNs jouent un rôle crucial en menant les ARN messagers cibles à la dégradation ou au blocage de leur traduction L'objectif de cette thèse est de comprendre et d'identifier la régulation du stress oxydatif en étudiant l'implication des micro-ARNs en réponse aux stress abiotiques et suite à l'apparition du TPD chez l'hévéa. L'isolement et le séquençage à haut débit de petits-ARNs ont permis l'identification de micro-ARNs d'hévéa. Dans un premier temps, une banque de petits ARNs a été effectuée à partir de vitroplants soumis ou non à des stress abiotiques, à laquelle s'est ajoutée dans un second temps deux banques fabriquées à partir de latex d'arbres en exploitation atteints ou non par le TPD. L'analyse de la population de petits ARNs montre une diminution de la taille des séquences en réponse à la maladie, la majorité des séquences de petits ARNs de latex étant de 21 nucléotides chez les arbres malades et 24 nucléotides chez les arbres sains. En combinant le pipeline LeARN et les données transcriptomiques, soixante huit familles de micro-ARNs conservés entre les espèces et quinze nouvelles familles de micro-ARNs ont été identifiées chez l'hévéa. Les gènes codant pour la voie de biogenèse des micro-ARNs sont présents dans le latex, suggérant leur production dans ce compartiment cellulaire particulier. L'identification des séquences de trente précurseurs de micro-ARNs ont permis d'étudier l'expression des gènes MIR en réponse aux stress abiotiques et en réponse au TPD. Les gènes MIR étudiés sont différentiellement exprimés chez des hévéas immatures en réponse au stress abiotiques et aux traitements par l'éthylène et le méthyl-jasmonate. L'abondance relative de transcrits des gènes MIR est fortement réduite par le TPD dès 5% de longueur d'encoche sèche à l'exception d'un gène.Les cibles potentielles des 83 familles de micro-ARNs ont été prédites. Ces micro-ARNs sont impliqués dans les voies de détoxication des espèces activées de l'oxygène, dans les voies de biosynthèse du caoutchouc naturel, dans les voies de biosynthèse et de signalisation de l'éthylène et du jasmonate. Trois cibles ont été validées expérimentalement dont la CuZnSOD chloroplastique, enzyme importante du système antioxydant. / Hevea brasiliensis is cultivated for natural rubber produced in latex cells. Intensive harvesting systems combined with environmental cues affect latex production. The Tapping Panel Dryness (TPD), a complex physiological disorder, causes a loss of production of 10-40%. Oxidative stress, starting point of the disease, affect latex flow because of in situ coagulation of rubber particles. In plants, the adaptation to abiotic stress relies on the fine tuning of the gene expression at the transcriptional and post-transcriptional levels. MicroRNAs play a crucial roles leading to mRNAs degradation or repression of their translation.The aim of this thesis is to understand and identify the regulation of the oxidative stress by studying the involvement of microRNAs in the regulation of abiotic stress and TPD occurrence in Hevea. Isolation and high-throughput sequencing of small RNAs allowed identifying Hevea microRNAs. Firstly, a small RNA library was constructed from in vitro plantlets subjected or not to abiotic stress, and secondly, two others small RNA libraries were constructed with latex from healthy and TPD-affected trees. Analyses of the small RNA population showed a decrease in the size of the reads in response to TPD, the majority of the small RNAs from latex being 21 nucleotides in TPD-affected trees and 24 nucleotides in healthy trees. Combining the LeARN pipeline and transcriptomic data, sixty eight microRNAs families conserved between plant species and fifteen new families were identified in Hevea. Genes involved in microRNA biogenesis are present in latex suggesting their production in this particular cellular compartment. Identification of thirty precursors of microRNAs allowed the expression analyses of the corresponding MIR genes in response to abiotic stress and upon TPD occurrence. MIR genes are differentially expressed in young plants in response to abiotic stress and in response to ethylene and methyl jasmonate treatments. Moreover, relative transcript abundance of MIR genes is strongly repressed upon TPD occurrence a soon as 5% of dry cut length except for one MIR gene.Putative targets were predicted for the 83 families. MicroRNAs are involved in ROS detoxification, natural rubber biosynthesis, ethylene and jasmonate biosynthesis and signalling pathways. Three targets were experimentally validated including the chloroplastic isoform of CuZnSOD, which is an important enzyme of the ROS-scavenging system.

Plants respond to environmental stress by altering their gene expression. Under stress conditions some genes are activated and some genes are repressed. Even though a lot of work has been done to understand mechanisms of gene activation under abiotic stress very little information is available on how stress responsive genes are kept repressed under normal growth conditions. Recent work has revealed that plants use transcriptional repression as common mechanism of gene repression. Transcriptional repression is achieved by recruitment co-repressor complexes to the target genes. Recent studies have revealed that the co-repressor LUH complexes with SLK1 and SLK2 to silence Arabidopsis thaliana stress responsive genes. However, the transcription factors involved in the recruitment of this complex to its target genes are not known. In this study, we identified SLK2INT1, as a novel transcription factor that is involved in silencing of select Arabidopsis thaliana stress responsive genes by recruiting the LUH-SLK2 complex.

The total annual forage production of tropical grasslands in the Brazilian Central area is expected to increase, however, some studies also predict a higher variability of forage production between and within years. In the future, maintenance of many pastures áreas will probably depend on the use of technology and increased productivity, seeking a competitive advantage concerning other activities, or even on its relocation to marginal áreas where forage grasses will be more suitable to abiotic factors. The water stress, both by flooding and drought, may reduce the production and survival of grasses. The objective was to identify the strategies that some genotypes of Paspalum and Urochloa genus use to cope with the flooding and/or water deficiency in the soil. Two greenhouse experiments were conducted: 1 - morphological and physiological characteristics, survival rate and recovery index in five genotypes of Paspalum sp. (BRS Guará, BGP 289, BGP 293, BGP 402 and BGP 397) and U. brizantha cv. Marandu were evaluated under three water conditions (flooded, drought and control); 2 - dehydration tolerance and dehydration avoidance of three cultivars of Urochoa (\'Marandu\', \'BRS Paiaguás\' and \'Basilisk\') and a cultivar Mediterranean Dactylis glomerata L. cv. Medly. In experiment 1, the genotypes of Paspalum sp., especially BGP 289, BGP 402, BGP 397 and cv. Marandu previously submitted water déficit, presented high tiller survival rates after rehydration. The cv. Marandu and BGP 289 access were not tolerant to flooding, presenting a low tiller survival rate and root recovery index. The BGP 293 access was not resistant to flooding, but it was able to survive periods up to 28 days under water depth. The accesses of Paspalum BRS Guará, BGP 402 and BGP 397 presented resistance mechanisms and survival to flooding, related to a high concentration of chlorophyll in the leaf, leaf biomass, and root recovery and tiller survival after a recovery period under water regular condition. In experiment 2, the distinction of cultivars was in accordance with the resource-use strategy. Cultivars of Urochloa were less tolerant to dehydration than cv. Medly. But the most productive Urochloa cultivars (Marandu and Basilisk) further delayed dehydration due to high investment in the root system, and consequently were less dehydration tolerant, with less survival rates under severe drought. The cv. BRS Paiaguás was less productive under regular irrigation than other Urochloa cultivars, but under drought conditions it is closer to the cv. Medly, being highly dehydration tolerant, and presenting high survival rates under severe drought and dehydration avoidance strategies with roots elongation along the soil profile during the drought period. There are differences in responses to other deficit and flooding between and within the Paspalum and Urochloa genus morphophysiological traits used in this study are relevant for the understanding of the mechanisms and discrimination among genotypes. / A partir de predições de futuros impactos climáticos, é esperado um aumento na produção anual total de forragem tropical, contudo estudos também predizem aumento da variação da produção entre e dentro dos anos. Diante desses cenários, é imprescindível buscar vantagens competitivas e o conhecimento do grau de tolerância a estresse abióticos de gramíneas tropicais. O estresses hídrico, tanto por excesso quanto por deficiência, pode reduzir a produção e a sobrevivência da planta forrageira. Assim, objetivou-se identificar as estratégias que alguns genótipos do gênero Paspalum e Urochloa utilizam para lidar com o encharcamento e/ou com a deficiência hídrica no solo. Foram conduzidos dois experimentos em casa-de-vegetação: 1 - avaliou-se características morfofisiológicas, taxa de sobrevivência e índice de recuperação em cinco genótipos de Paspalum sp. (BRS Guará, BGP 289, BGP 293, BGP 402 e BGP 397) e U. brizantha cv. Marandu sob três condições hídricas (alagado, déficit e controle), em vasos pequenos; 2 - mensurou-se em pequenos vasos a tolerância a desidratação e em longos tubos o atraso a desidratação de três cultivares de Urochoa (\'Marandu\', \'BRS Paiaguás\' e \'Basilisk\') e uma cultivar mediterrânea de Dactylis glomerata L. cv. Medly, submetidos a condições sem limitação hídrica e posteriormente sob seca prolongada. Sob déficit hídrico, os genótipos de Paspalum sp., especialmente o BGP 289, o BGP 402 e o BGP 397, e a cv. Marandu, apresentaram altas taxas de sobrevivência de perfilhos após reidratação. A cv. Marandu e o acesso BGP 289 não são tolerantes ao alagamento, apresentaram baixas taxas de sobrevivência e de recuperação radicular. O acesso BGP 293 não é resistente ao alagamento, porém é capaz de sobreviver a períodos de até 28 dias sob lâmina d\'água. Os acessos de Paspalum BRS Guará, BGP 402 e BGP 397 apresentam mecanismos de resistência e sobrevivência ao alagamento, relacionados à alta concentração de clorofila na folha, de biomassa de folhas, e de recuperação radicular e sobrevivência de perfilhos após período de recuperação sob condição hídrica regular. No experimento 2, a distinção dos cultivares foi de acordo com a estratégia de uso dos recursos. Cultivares de Urochloa são menos tolerantes a desidratação que a cv. Medly. Mas as cultivares de Urochloa mais produtivas (Marandu e Basilisk) atrasaram mais a desidratação, devido ao alto investimento no sistema radicular, e consequentemente foram menos tolerantes a desidratação, com memores taxas de sobrevivência sob seca severa. A cv. BRS Paiaguás é menos produtiva sob irrigação regular em comparação as outras cultivares de Urochloa, mas sob seca aproximou-se mais da cv. Medly, mostrando-se altamente tolerante a desidratação, com altas taxas de sobrevivência sob seca severa, além de apresentar estratégia de atraso a desidratação com o alongamento de raízes ao longo do perfil do solo durante a seca. Existem diferenças entre e dentro de espécies dos gêneros Paspalum e Urochloa em relação as respostas sob duas formas de estresse hídrico. As caraterísticas morfofisiológicas utilizadas neste estudo auxiliaram na compreensão dos mecanismos de resposta ao estresse hídrico e na discriminação dos genótipos.

Os objetivos deste trabalho com Mahanarva fimbriolata foram estudar a influência de fatores abióticos (temperatura, umidade e fotoperíodo) sobre sua biologia, avaliar a flutuação populacional, sistema de amostragem de ninfas e adultos e os métodos de controle, biológico com o fungo Metarhizium anisopliae, químico com thiamethoxam (Actara® 250WG) e por resistência varietal. As condições térmicas influem na duração das fases do ciclo, não havendo desenvolvimento embrionário a 32°C. As temperaturas constantes de 30 e 32°C são letais para ninfas, sendo 25°C a temperatura ideal. Os limites térmicos inferiores de desenvolvimento e as constantes térmicas são 11,2°C e 318,2 graus-dias para a fase de ovo e 11,8°C e 468,0 graus-dias para a fase de ninfa. Condições térmicas flutuantes influem na duração das fases do inseto, mas não são letais a nenhuma delas. A umidade influi na duração e viabilidade do período embrionário. O menor período ocorre na umidade de contato, seguido pela UR de 100%. Nas UR de 80, 60 e 40%, não há desenvolvimento embrionário. Ovos expostos a essas umidades por 120 dias e, então, submetidos à umidade de contato, continuam inviáveis ou com baixas viabilidades (9,6; 6,3 e 0%, respectivamente). Indivíduos expostos à fotofase de 10 horas colocam ovos inférteis ou em aparente diapausa. Para sugarem a seiva, as ninfas inserem os estiletes na epiderme da raiz, atravessam todo o córtex e atingem o cilindro vascular, alimentando-se no xilema e no tubo crivado do floema primário. Na lâmina foliar, os adultos introduzem os estiletes, preferencialmente, pelos estômatos, atravessam as células do parênquima clorofiliano e atingem o metaxilema nos feixes vasculares. O aparecimento da cigarrinha está condicionado ao excedente hídrico e à temperatura. É possível se amostrarem adultos, com armadilhas adesivas amarelas e determinar o momento para controlar ninfas de mesma geração. Não há diferença em relação ao número de conídios depositados por ninfa em concentrações que variam de 1,6 a 5,3; 2,9 a 4,9 e 0,7 a 3,6 conídios/ninfa, respectivamente. As maiores mortalidades acumuladas, total e confirmada, de M. fimbriolata pulverizada com M. anisopliae, oito dias após a inoculação, são constatadas na dosagem de 5,0 x 108 conídios.ml-1, sendo de 57,5 e 62,3% (total) e 42,7 e 45,2% (confirmada) com pulverização sobre ninfa e espuma, respectivamente. M. anisopliae apresenta maior potencial para controle da cigarrinha quando pulverizado sobre a ninfa ou a espuma e sem a lavagem de conídios após a aplicação. Dentre as variedades estudadas, SP81-3250 é a mais suscetível à cigarrinha, possibilitando maior longevidade de machos e fêmeas, maior período de oviposição e elevada fecundidade. A variedade RB72454 reduz em 50% a população de ninfas nas raízes. A aplicação de thiamethoxam, a 1,0 kg p.c./ha, na base das plantas, proporciona controle superior a 90% até 60 DAA e residual de 105 dias (0,02 mg.kg-1) nas raízes. A eficiência de controle na pulverização foliar diminui para 34%, 15 DAA, com resíduos de 2,61 mg.kg-1 nas raízes. Pulverizações dirigidas ao colmo apresentam resíduos inferiores a 0,02 mg.kg-1 nas raízes, sem controle da praga. / The objectives of this work were to study the influence of abiotic factors (temperature, moisture, and photoperiod) on Mahanarva fimbriolata biology, and to evaluate population fluctuation, sampling system for nymphs and adults, and biological control with the fungus Metarhizium anisopliae, chemical control with thiamethoxam (Actara® 250 WG), and control by varietal resistance. The temperature conditions influence life cycle stage duration; no embryonic development is observed at 32°C. Constant temperatures of 30 and 32°C are lethal to nymphs, whereas 25°C is the ideal temperature. The lower temperature development thresholds and thermal constants are 11.2°C and 318.2 degree-days for the egg stage and 11.8°C and 468.0 degree-days for the nymphal stage. Fluctuating temperature conditions influence stage duration but are not lethal to any of the stages. Humidity influences the duration and viability of the embryonic period. The shortest period occurs at contact humidity, followed by 100% RH No embryonic development occurs at RH values of 80, 60, and 40%. Eggs exposed to these humidity values for 120 days and then submitted to contact humidity remain unviable or present low viability (9.6; 6.3, and 0%, respectively). Individuals exposed to a 10-hour photophase lay infertile or apparently-diapausing eggs. In order to suck sap, the nymphs insert their stylets into the root epidermis, passing through the entire cortex and reaching the vascular cylinder, feeding on the xylem and on the sieve tube of the primary phloem. On the leaf blade, the adults introduce their stylets preferentially through the stomata, passing through the chlorophyll-bearing parenchyma cells and reaching the metaxylem on the vascular bundles. Spittlebug occurrence is conditioned to water surplus and temperature. Adults can be sampled by means of yellow sticky traps, allowing the control of same-generation nymphs to be synchronized. There are no differences with regard to the number of conidia deposited per nymph at concentrations ranging from 1.6 to 5.3; 2.9 to 4.9; and 0.7 to 3.6 conidia/nymph, respectively. The highest cumulative, total, and confirmed mortalities of M. fimbriolata sprayed with M. anisopliae eight days after inoculation are verified at a dose of 5.0 × 108 conidia.ml-1, with values of 57.5 and 62.3% (total), and 42.7 and 45.2% (confirmed) when sprayed on the nymphs and on the spittle, respectively. M. anisopliae has a greater potential to control spittlebugs when sprayed on nymphs or on the spittle without washing off the conidia after application. Among the varieties studied, SP81-3250 is the most susceptible to the spittlebug, allowing greater longevity of males and females, a longer oviposition period, and high fecundity. Variety RB72454 reduces the nymphal population on roots by 50%. The application of thiamethoxam at 1.0 kg c.p./ha at the base of plants provides control rates higher than 90% until 60 DAA and a residual control of 105 days (0.02 mg.kg-1) on the roots. Leaf spray control effectiveness decreases to 34% at 15 DAA, with residues of 2.61 mg.kg-1 in roots. Sprays directed at the stalk show residues lower than 0.02 mg.kg-1 in roots, without controlling the pest.

Low bioavailability of phosphate (P) due to low concentration and high immobility in soils is a key limiting factor in crop production. Application of excess amounts of P fertilizer is costly and by no means sustainable, as world-wide P resources are finite and running out. To facilitate the breeding of crops adapted to low-input soils, it is essential to understand the consequences of P deficiency. The second messenger calcium (Ca2+) is known to signal in plant development and stress perception, and most recently its direct role in signalling nutrient availability and deficiency has been partially elucidated. The use of Ca2+ as a signal has to be tightly controlled, as Ca2+ easily complexes with P groups and therefore is highly toxic to cellular P metabolism. It is unknown whether Ca2+ signals P availability or whether signalling is altered under P starvation conditions. The aim of this PhD project was to characterise the use of Ca2+ ions, particularly cytosolic free Ca2+ ([Ca2+]cyt), in stress signalling by P-starved roots of the model plant Arabidopsis thaliana. The hypothesis was that under P starvation and a resulting decreased cellular P pool, the use of [Ca2+]cyt may have to be restricted to avoid cytotoxic complexation of Ca2+ with limited P groups. Employing a range of genetically encoded Ca2+ reporters in Arabidopsis, P starvation but not nitrogen starvation was found to strongly dampen the root [Ca2+]cyt increases evoked by mechanical, salt, osmotic, and oxidative stress as well as by extracellular nucleotides. The strongly altered root [Ca2+]cyt response to extracellular nucleotides was shown to manifest itself during seedling development under chronic P deprivation, but could be reversed by P resupply. Fluorescent imaging elucidated that P-starved roots showed a normal [Ca2+]cyt response to extracellular nucleotides at the apex, but a strongly dampened [Ca2+]cyt response in distal parts of the root tip, correlating with high reactive oxygen species (ROS) levels induced by P starvation. Excluding iron, as well as P, rescued the altered [Ca2+]cyt response, and restored ROS levels to those seen under nutrient-replete conditions. P availability was not signalled through [Ca2+]cyt. In another part of this PhD project, a library of 77 putative Ca2+ channel mutants was compiled and screened for aberrant root hair growth under P starvation conditions. No mutant line showed aberrant root hair growth. These results indicate that P starvation strongly affects stress-induced [Ca2+]cyt modulations. The data generated in this thesis further understanding of how plants can integrate nutritional and environmental cues, adding another layer of complexity to the use of Ca2+ as a signal transducer.

O trigo (Triticum aestivum L.) é uma importante cultura no Brasil. Poucas cultivares são recomendadas para produção do tipo sequeiro no Bioma Cerrado onde a escassez de água limita o rendimento de grãos. Aqui reportamos uma análise de transcriptoma do MGS1 Aliança (cultivar de trigo adaptada ao Cerrado) sob estresse de seca. Um grupo de 4.422 transcritos diferencialmente expressos foi encontrado em raízes e folhas. O número de transcritos reprimidos em raiz (1.102) foi menor que os transcritos induzidos (1.706), enquanto o oposto ocorreu em folhas (1,017 induzidos e 647 reprimidos). O número de transcritos comuns entre ambos órgaõs foi 1.249, enquanto 2.124 foram específicos para raíz e 1.049 específicos para folhas. Análises de RT-qPCR de 35 transcritos selecionados ao acaso revelou uma correlação de 0,78 com os dados de transcriptoma. Os transcritos diferencialmente expressos foram distribuídos por todos os cromossomos e componentes do genoma. O número de transcritos no genoma B foi maior do que nos genomas A e D. Ainda, um grande número de transcritos relacionados à seca foi mapeado nos cromossomos 3B, 5B e 2B. Quando consideramos ambos órgãos, 116 diferentes rotas metabólicas foram alteradas. Uma rota em comum, entre as três mais alteradas em ambos órgãos, foi o metabolismo do amido e da sacarose. A comparação de transcritos derivados de raiz e de folha permite a identificação de transcritos importantes relacionados à respota ao estresse de seca em cada um destes órgãos. Os dados obtidos, também, abrem caminho para o desenvolvimento de futuros marcadores e seleção de genes candidatos ligados à característica. Estes resultados são úteis para o entendimento de rotas metabólicas envolvidas na tolerância à seca em trigo. A informação gerada será usada, a mais longo prazo, para propósitos de transgenia. Para isto, a metodologia de duplo-haploides é desejável e uma primeira investigação sobre a eficiência de protocolo se mostrou necessária. Micrósporos são células gaméticas com capacidade de dar origem a uma nova planta via embriogênese in vitro. Plantas duplo-haploides geradas pela cultura de micrósporos isolados são completamente homozigotas e representam uma importante ferramenta para estudos genéticos e melhoramento de plantas O processo androgenético é desencadeado por diferentes pré-tratamentos de estresse, os quais são empregados para mudar os micrósporos da rota gametofítica para a rota esporofítica. Embora a cultura de micrósporos isolados tenha inúmeras vantagens, importantes limitações tem impedido sua apliação em larga escala. Diferenças genotípicas na resposta androgenética e na formação de plantas albinas ainda constituem desafios. Embora o albinismo seja principalmente uma característica genética, pré-tratamentos e meios de cultura apropriados podem evitar este fenômeno até certo ponto. A resposta androgenética de cinco genótipos de trigo brasileiro foi avaliada no presente estudo. Dois pré-tratamentos foram testados: frio (4°C) e ácido 2-hidroxinicotinico (100 mg/L). O frio foi melhor que o pré-tratamento químico, produzindo mais plantas verdes em quatro de cinco genótipos. Somente dois genótipos brasileiros tratados com ácido 2-hidroxinicotinico produziram plantas, e um deles apenas uma única planta albina. Nossos reultados mostram, também, que o meio semilíquido (contendo 10% de Ficoll) promoveu uma maior resposta androgenética que o meio líquido, aumentando o número de embriões e plantas regeneradas. / Wheat (Triticum aestivum L.) is an important crop cultivated in Brazil. Few cultivars are recommended for rainfed production in the Cerrado Biome where water scarcity limits grain yield. Here we report a transcriptome analysis of MGS1 Aliança (a wheat cultivar adapted to the Cerrado) under drought stress. A set of 4,422 differentially expressed transcripts was found in roots and leaves. The number of down-regulated transcripts in roots (1,102) was lower than the up-regulated transcripts (1,706), while the opposite occurred in leaves (1,017 induced and 647 repressed). The number of common transcripts between the two tissues was 1,249, while 2,124 were specific to roots and 1,049 specific to leaves. Quantitative RT-PCR analysis of 35 randomly selected transcripts revealed a 0.78 correlation with the transcriptome data. The differentially expressed transcripts were distributed across all chromosomes and component genomes. The number of transcripts on the B genome was greater than on the A and D genomes. Additionally, a greater number of drought related transcripts was mapped on chromosomes 3B, 5B and 5D. When considering both tissues, 116 different metabolic pathways were changed. One common pathway, among the top three changed pathways in both tissues, was starch and sucrose metabolism. The comparison of root- and leaf-derived transcripts allows the identification of important transcripts related to water stress response in each of these tissues. It also paves the way for future marker development and selection of candidate genes linked to that trait. These results are useful for understanding the metabolic pathways involved in wheat drought response. The information generated will be used for transgenic wheat purposes. For this the doubled-haploid method is desirable and an investigation about the protocol eficiency is needed. Microspores are gametic cells with capacity to give rise to a new plant via in vitro embryogenesis. Doubled haploid plants generated by isolated microspore culture are completely homozygous and represent an important tool for plant genetics and breeding research. This process is triggered by different stress pretreatments, which are employed to switch microspores from gametophytic to a sporophytic pathway. Although isolated microspore culture has innumerous advantages, important limitations have prevented its application on a large scale. Genotypic differences in androgenic response and the formation of albino plants remain great challenges. Although albinism is a major genetic characteristic, appropriated pretreatments and culture medium can avoid this phenomenon to some extent. The androgenic response of five Brazilian wheat genotypes was evaluated in the present study. Two pretreatments were tested: cold (4°C) and 2-hydroxynicotinic acid (100 mg/L). Cold was better than chemical pretreatment, producing more green plants in four out of five genotypes. Only two Brazilian genotypes treated with 2-hydroxynicotinic acid produced plants, and one of them produced a single albino plant. Our results also show that semi-liquid medium (containing 10% Ficoll) promoted a higher androgenic response than did liquid medium, increasing the number of embryos and regenerated plants.

Submitted by Maria Beatriz Vieira (mbeatriz.vieira@gmail.com) on 2017-03-29T13:15:52Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) dissertacao_vania_marques_gehling.pdf: 823705 bytes, checksum: bb9a04c4074928df8353da917cbc35a4 (MD5) / Approved for entry into archive by Aline Batista (alinehb.ufpel@gmail.com) on 2017-04-05T17:44:43Z (GMT) No. of bitstreams: 2 dissertacao_vania_marques_gehling.pdf: 823705 bytes, checksum: bb9a04c4074928df8353da917cbc35a4 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Aline Batista (alinehb.ufpel@gmail.com) on 2017-04-05T17:45:44Z (GMT) No. of bitstreams: 2 dissertacao_vania_marques_gehling.pdf: 823705 bytes, checksum: bb9a04c4074928df8353da917cbc35a4 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2017-04-05T17:45:54Z (GMT). No. of bitstreams: 2 dissertacao_vania_marques_gehling.pdf: 823705 bytes, checksum: bb9a04c4074928df8353da917cbc35a4 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2015-09-03 / Sem bolsa / O objetivo deste trabalho foi avaliar o desempenho fisiológico de sementes e plântulas de pimenta e de tomateiro sob influência de diferentes temperaturas. Foram conduzidos, separadamente, dois experimentos no Laboratório Didático de Análise de Sementes – FAEM - UFPel. No primeiro experimento foram utilizadas sementes de pimenta Cayenne Long Slim®, e no segundo experimento foram empregadas sementes de tomateiro Gaúcho®. Em ambos os experimentos, os tratamentos consistiram de temperaturas constantes, sendo: 18; 20; 22; 24 e 26ºC. Para a determinação do desempenho fisiológico das sementes e plântulas foram empregadas as avaliações de germinação, primeira contagem de germinação, índice de velocidade de germinação, envelhecimento acelerado, teste de frio, comprimento de parte aérea e de raiz primária, massa da matéria seca de parte aérea e de raízes, emergência de plântulas aos 14 e 21 dias após a semeadura e índice de velocidade de emergência. Os resultados demonstraram que a germinação de sementes de pimenta não é afetada de maneira marcante pelas temperaturas de 18 a 24oC, assim como sementes de tomateiro sob temperatura de 18 a 26oC, que não apresentaram diferença significativa na germinação. A primeira contagem de germinação, o índice de velocidade de germinação e os valores de envelhecimento acelerado foram incrementados linearmente com o aumento da temperatura, em pimenta e tomateiro. Todavia, o teste de frio e de emergência, tanto aos 14 como aos 21 dias após a semeadura, foram influenciados positivamente, até a temperatura aproximada de 22ºC, para ambas as culturas. Deste modo, a expressão do vigor de sementes, de ambas as espécies, é afetada negativamente pela redução da temperatura de desenvolvimento, sendo resultados marcantes obtidos nas temperaturas de 18 e 20oC. / The objective of this study was to evaluate the physiological performance of seeds and pepper and tomato seedlings under the influence of different temperatures. They were conducted separately in two experiments in the Didactic Laboratory of Seed Analysis - FAEM - UFPel. In the first experiment pepper seeds were used Cayenne Long Slim®, and in the second experiment Gaúcho® tomato seeds were employed. In both experiments, the treatments consisted of constant temperatures, as follows: 18; 20; 22; 24 and 26ºC. To determine the physiological performance of the seeds and seedlings were employed germination assessments, first count, germination speed index, accelerated aging, cold test, shoot length and primary root mass of dry matter air and root, seedling emergence at 14 and 21 days after sowing and emergence speed index. The results show that the pepper seed germination is not affected markedly by the temperature from 18 to 24ºC, as well as tomato seeds at a temperature of 18 to 26ºC, which showed no significant difference in germination. The first count, germination speed index and the accelerated aging values were increased linearly with increasing temperature in pepper and tomato. However, the cold test and emergency, both at 14 and at 21 days after sowing, were influenced positively to the approximate temperature of 22°C, for both crops. Thus, the expression of seed vigor of both species is adversely affected by reducing the growth temperature, with remarkable results obtained at temperatures of 18 to 20ºC.