<p>  </p> <p>Giant ragweed (<em>Ambrosia trifida</em> L.) is an annual broadleaf plant capable of emergence throughout the cropping season, opportune colonization of disturbed soil, rapid biomass accumulation, and a propensity to evolve mutations that endow resistance to herbicides, all of which contribute to giant ragweed being one of the most challenging weeds to control in row-crop production. Many soybean growers rely on acetolactate synthase (ALS)-inhibiting herbicides such as cloransulam for control of giant ragweed prior to its emergence, though the spread of biotypes resistant to ALS inhibitors can render these herbicides largely ineffective. Mesotrione inhibits the 4-hydroxyphenylpyruvate dioxygenase (HPPD) enzyme, and applications of this herbicide have recently been approved for use in mesotrione-resistant soybean varieties. Field experiments demonstrated that preemergence applications of mesotrione resulted in greater control of giant ragweed populations segregating for ALS-inhibitor resistance than several other commonly used herbicide combinations. Where mesotrione was applied, giant ragweed biomass was reduced by an average of 84% relative to the nontreated, while treatments without mesotrione increased biomass by an average of 34% by suppressing competition from other weed species. Additionally, both soil- and agar-based bioassays demonstrated that combinations of mesotrione and metribuzin can be synergistic for control of giant ragweed. </p> <p>Cloransulam was shown to result in strong selection for giant ragweed individuals with ALS-inhibitor resistance, increasing the proportion of resistant plants that emerged at one field site from 15% to greater than 90% after a single preemergence application. This selection pressure was reduced when mesotrione was co-applied with cloransulam. However, no herbicide combination, including sequential applications of non-ALS-inhibiting herbicides, consistently resulted in a resistance frequency similar to the baseline if an ALS inhibitor was applied preemergence. Resistance to cloransulam and other ALS inhibitors is expressed in giant ragweed plants possessing at least one mutant (Trp574Leu) <em>ALS</em> allele. The distribution of this allele in one field violated the Hardy-Weinberg Equilibrium, despite the fact that <em>ALS</em> is a nuclear gene and the Trp574Leu mutation does not incur a fitness penalty. We suspected that the inheritance of this mutation may be linked with a gene or genes responsible for self-incompatibility (SI) in giant ragweed, and that linkage drag was disrupting pollination in resistant plants. This research provided evidence that giant ragweed does possess SI, as greater pollen retention, pollen tube growth, and seed set were observed in cross-pollinated plants compared with self-pollinated plants. Non-Mendelian inheritance of the Trp574Leu mutation was documented in crosses between plants from three different giant ragweed populations, indicating that the mutant <em>ALS</em> allele may be linked with an SI allele common to many plants because of a shared resistant ancestor.  In crosses between plants from one population, production of resistant F1 seeds was 33% greater on average compared with the expectation under Mendelian inheritance. </p> <p>Collectively, this research demonstrated that mesotrione may become a highly effective tool for control of giant ragweed in soybean. Applications of mesotrione can also reduce the selection for an increased frequency of ALS inhibitor-resistant biotypes induced by cloransulam, though a more robust weed management strategy may be necessary to maintain the long-term viability of ALS inhibitors.  The need for sound weed management practices is underscored by the impact of the linkage of SI and <em>ALS</em> genes, which may be encouraging a more rapid spread of herbicide-resistance than was previously anticipated.</p>

No description available.

The genus Restrepia is well known to orchid enthusiasts but its micromorphology has not been described, and its pollination and breeding systems have not been investigated. The aim of this investigation was, therefore, to add to existing knowledge so that the resultant data could be used to facilitate ex situ conservation initiatives. A detailed electron microscopy study (SEM) of the floral organs was performed. This confirmed the structure of the dorsal sepal and lateral petal osmophores, their secretory nature together with that of the synsepal and the labellum. It was postulated how, by manipulating different labellar surface textures, the flower might use these ‘tactile guides’ to steer the insect (fly) through the flower. The cirrhi were postulated to help by destabilising the pollinator in flight, trapping it and bringing about pollination. The papillate structure of the calli was established and their optical properties investigated. Media comparison investigations established that Western medium supported the highest germination rates and, with the addition of banana supplement, the highest rates for seedling growth and development. This represented the first protocol for axenic germination of Restrepia in the literature (Millner et al., 2008) and provided a tested methodology for investigating breeding systems and producing Restrepia plant material for both scientific and horticultural purposes. Self-pollinations were found to produce fewer embryos compared to cross-pollinations. The operation of self-incompatibility (SI) was confirmed by the study of pollen tube growth which further confirmed the time interval between pollination and fertilisation. A time line from pollination/fertilisation to flowering was established. The type of SI in operation was best explained by gametophytic incompatibility. This demonstrated that it was possible to raise Restrepia hybrids and species from seed, by performing intraspecific crosses so helping to preserve them for posterity and relieve pressure on wild populations. Narrow endemic Restrepia species face combined threats from habitat loss, habitat degradation and problems of viable seed production due to the effects of SI and inbreeding depression (ID). Recently developed online resources, such as GeoCAT, were used to perform a Red List assessment in order to identify the degree of threat individual species faced, both globally and nationally. All species were classified as facing substantial levels of threat; although this was lessened for populations in protected habitats. Conservation is needed for cultivated collections as well as these wild populations by keeping alive existing knowledge and expertise in growing these species.

Depuis les années 1950, l’intensification des pratiques agricoles concourt à une augmentation de la pression anthropique entraînant une raréfaction des espèces spécialistes des parcelles cultivées, ce qui contribue à l’érosion de la diversité biologique des agroécosystèmes. Parmi les espèces en déclin, on compte de nombreuses messicoles dont le bleuet (Centaurea cyanus L.), fleur emblématique des moissons. Or, cette espèce peut rendre des services écosystémiques comme hôte de prédateurs de ravageurs des cultures et ressource privilégiée de certains pollinisateurs qui justifieraient son maintien dans les champs. Nous étudions ici des facteurs biologiques qui pourraient potentiellement contribuer à son déclin : distribution spatiale, potentialité de croissance, diversité génétique des populations, survie des semences, système de reproduction. Nous avons montré, par deux campagnes de relevés, que C. cyanus n’est pas une espèce indicatrice de diversité floristique dans la parcelle cultivée. Toutefois, lorsque que le bleuet est rare dans une région, il est associé à d’autres messicoles peu fréquentes. Par contre, s’il est commun, il est associé aux zones ayant la plus forte diversité végétale. Une expérience de semis dans différents compartiments de l’agrosystème, en absence de traitements herbicides, a mis en évidence que la potentialité de croissance des bleuets est plus élevée dans le plein champ du blé que dans la moutarde et les interfaces blé/bordure. Cette croissance est limitée de manière variable par les communautés adventices présentes dans les compartiments hors champs (bordures). L’analyse de la diversité génétique à l’aide de marqueurs microsatellites de bleuets dans une petite zone agricole montre que les populations sont connectées par des flux de gènes importants. Les barrières écologiques telles que les chemins, semblent être des facteurs de structuration plus importants que la distance géographique séparant les populations. La répartition du bleuet dans le paysage agricole n’est donc pas aléatoire et apparaît dépendante de la fréquence de l’espèce dans la région ainsi que des différents éléments du paysage. Les caractéristiques du cycle biologique du bleuet ont été étudiées grâce à des expériences au champ et en serre. Nous avons montré que la longévité des akènes enfouis dans le sol chute rapidement après deux ans. Le cycle saisonnier de la dormance permet deux cohortes de levées (automne et printemps). L’étude du système de reproduction a permis de mettre en évidence que les pollinisateurs sont nécessaires pour la fécondation et que les populations sont majoritairement auto-incompatibles. Il existe cependant des individus pseudo auto-incompatibles mais leur fréquence n’est pas liée à la taille des populations ou à leur niveau d’isolement spatial. Par ailleurs, dès que le coefficient de consanguinité augmente dans les populations, la valeur phénotypique des individus baisse. La dépression de consanguinité s’exprime alors principalement pendant la phase de la germination. L’ensemble de ces caractéristiques biologiques peuvent s’avérer défavorables au bleuet dans les agroécosystèmes simplifiés actuels et ont pu entraîner son déclin dans certaines régions / Agroecosystems are currently experiencing high biodiversity loss, in particular among the plant species specifically adapted to this habitat. This decline results from cropping systems that have been intensified in Western Europe since the 1950s. The cornflower (Centaurea cyanus L.), considered as emblem of the flora associated with traditional cereals, appears as a species that may be at risk and should be monitored. Indeed, cornflower can serve as a host to predators of crop pests and is strongly attractive for the pollinators. This study examines the biological factors that could potentially cause the decline of this species: spatial distribution, potential for growth, population’s genetic diversity, seeds survival and mating system.We have shown that C. cyanus is not a biological indicator of the weed diversity of a field. However, when it is not frequent in a region, cornflower is associated with other rare segetal species. When it is common in a region, C. cyanus is present in areas with higher weed diversity. Sowing seeds in different components of the agrosystem, without herbicide application, shows that the growth of cornflower was higher in the centre of a wheat field than in mustard and the crop edge. Growth success in field margins was variably limited according to the weed communities in the field boundaries. The analysis of the genetic diversity using microarrays has shown that populations in a small agricultural area are connected by important genes flow. The ecological barriers as country road seem to be factors more determinant than geographical distance to structure and divide populations. Therefore, the cornflower distribution in the agricultural landscape is not random and appears to be linked to species frequency in the area and to landscape elements.The cornflower life cycle characteristics were studied through field and greenhouse experiments. We have shown that the longevity of achenes buried in the soil falls rapidly after two years. There is a seasonal dormancy cycle that allows the emergence of two seedling cohorts (fall and spring). Our study of the mating system highlights the fact that pollinators are required for fertilization and that cornflower is strongly self-incompatible. However, there are pseudo self-incompatible individuals, but their frequency is not related to the population size or the spatial isolation level. Individual fitness decreases as the inbreeding coefficient increases in a population. Inbreeding depression is mainly expressed during the germination stage. All these biological characteristics can be detrimental to the cornflower in current simplified agroecosystems and may have led to cornflower decline in some areas

<p>In this thesis, I examined variation in sex expression and mating patterns in the sexually polymorphic, wind-pollinated herb <i>Plantago maritima</i>. With a combination of field studies, greenhouse experiments, and genetic analyses, I (a) examined factors influencing sex ratio variation in gynodioecious plants (in which hermaphrodites and females coexist), (b) discovered variation in breeding system, (c) investigated density-dependence of seed production, and (d) documented genetic variation within and among populations close to the northern range margin in Europe. </p><p>In a survey of 104 <i>P. maritima</i> populations, I documented considerable variation in sex ratio (range 0-70% females, median 6.3% females). As predicted, females were more frequently missing from small than from large populations, and the variance in sex ratio increased with decreasing population size. Among twelve populations sampled for seed production, the frequency of females was positively related to relative fecundity of females and negatively related to population size. The results suggest that the local sex ratio is influenced both by the relative fecundity of females and hermaphrodites, and by stochastic processes in small populations.</p><p>A comparative field study showed that plant fecundity decreased with increasing distance to nearest pollen donor both within and among populations in an archipelago in southern Sweden, where self-incompatibility was confirmed in controlled crosses. In contrast, plant fecundity was overall higher and was not density-dependent in the Skeppsvik archipelago in northern Sweden, where controlled crosses showed that plants are self-compatible. The results were consistent with the prediction that evolution of self-fertility should reduce density-dependence of pollination success.</p><p>I quantified the genetic structure within and among populations from eastern Sweden and western Finland based on variation at four polymorphic microsatellite loci. The genetic diversity was low in northern Sweden, which may be the result of a history of small population sizes and periods of frequent self-fertilization.</p>

In this thesis, I examined variation in sex expression and mating patterns in the sexually polymorphic, wind-pollinated herb Plantago maritima. With a combination of field studies, greenhouse experiments, and genetic analyses, I (a) examined factors influencing sex ratio variation in gynodioecious plants (in which hermaphrodites and females coexist), (b) discovered variation in breeding system, (c) investigated density-dependence of seed production, and (d) documented genetic variation within and among populations close to the northern range margin in Europe. In a survey of 104 P. maritima populations, I documented considerable variation in sex ratio (range 0-70% females, median 6.3% females). As predicted, females were more frequently missing from small than from large populations, and the variance in sex ratio increased with decreasing population size. Among twelve populations sampled for seed production, the frequency of females was positively related to relative fecundity of females and negatively related to population size. The results suggest that the local sex ratio is influenced both by the relative fecundity of females and hermaphrodites, and by stochastic processes in small populations. A comparative field study showed that plant fecundity decreased with increasing distance to nearest pollen donor both within and among populations in an archipelago in southern Sweden, where self-incompatibility was confirmed in controlled crosses. In contrast, plant fecundity was overall higher and was not density-dependent in the Skeppsvik archipelago in northern Sweden, where controlled crosses showed that plants are self-compatible. The results were consistent with the prediction that evolution of self-fertility should reduce density-dependence of pollination success. I quantified the genetic structure within and among populations from eastern Sweden and western Finland based on variation at four polymorphic microsatellite loci. The genetic diversity was low in northern Sweden, which may be the result of a history of small population sizes and periods of frequent self-fertilization.

Compatible interactions between male gametophytes (pollen) and the female reproductive organ (pistil) are essential for fertilization in flowering plants. Recognition at a molecular level allows “compatible” pollen grains to adhere/germinate on the stigma while pollen grains from unrelated plant species are largely ignored. The exocyst is a large eight subunit complex that is primarily involved in polarized secretion or regulated exocytosis in eukaryotic cells where it functions to tether vesicles to the plasma membrane. Recent research has implicated one of the Exo70 family members, Exo70A1, in compatible pollen-pistil interactions in Arabidopsis and Brassica. The loss of Exo70A1 in Arabidopsis Col-0 stigmas leads to the rejection of compatible pollen producing a “female sterile” phenotype. Through my research I have demonstrated that, driven by a stigma-specific promoter, an RFP:Exo70A1 fusion protein rescues this defect in exo70A1-1 mutant and Exo70A1 is found to be localized to the plasma membrane at flower opening.

Compatible interactions between male gametophytes (pollen) and the female reproductive organ (pistil) are essential for fertilization in flowering plants. Recognition at a molecular level allows “compatible” pollen grains to adhere/germinate on the stigma while pollen grains from unrelated plant species are largely ignored. The exocyst is a large eight subunit complex that is primarily involved in polarized secretion or regulated exocytosis in eukaryotic cells where it functions to tether vesicles to the plasma membrane. Recent research has implicated one of the Exo70 family members, Exo70A1, in compatible pollen-pistil interactions in Arabidopsis and Brassica. The loss of Exo70A1 in Arabidopsis Col-0 stigmas leads to the rejection of compatible pollen producing a “female sterile” phenotype. Through my research I have demonstrated that, driven by a stigma-specific promoter, an RFP:Exo70A1 fusion protein rescues this defect in exo70A1-1 mutant and Exo70A1 is found to be localized to the plasma membrane at flower opening.

Self-incompatibility is a genetic barrier by which a plant recognizes and rejects its own pollen while allowing pollen from more distantly related plants to germinate. In the Brassicacea family, it is controlled by a highly polymorphic locus called the S-locus, which contains the male and female determinants of self-incompatibility. The stigma expresses the female determinant of self-incompatibility, the plant receptor kinase (PRK) S-LOCUS RECEPTOR KINASE (SRK). In Brassica oleracea, SRK has a unique subcellular localization among PRK: the receptor is mostly localized in endosomes and to a lesser extent at the plasma membrane.We investigated the function of the endosomal localization of SRK in Arabidopsis thaliana. Firstly, we reintroduced self-incompatibility in Arabidopsis thaliana by expression of a functional SRK allele from Arabidopsis lyrata (a self-incompatible species). Secondly, we showed that a loss-of-function mutant of DYNAMIN-RELATED PROTEIN1A, a protein required for endocytosis, abolished self-incompatibility. Our results suggest that endocytosis is required for self-incompatibility, and that SRK may be signaling from endosomal compartments.

Submitted by Claudia Rocha (claudia.rocha@udesc.br) on 2018-03-01T12:12:19Z No. of bitstreams: 1 PGPV17MA218.pdf: 1720230 bytes, checksum: 6b5c28d9052e3150143609b5794ee96a (MD5) / Made available in DSpace on 2018-03-01T12:12:19Z (GMT). No. of bitstreams: 1 PGPV17MA218.pdf: 1720230 bytes, checksum: 6b5c28d9052e3150143609b5794ee96a (MD5) Previous issue date: 2017-02-02 / PROMOP / CAPES / Due to the gametophytic self-incompatibility (GSI), the occurrence of cross-pollination between genetically compatible plants is necessary for the naturally fructification of apple trees (Malus domestica Borkh.). The GSI is governed by the multiallelic locus called S, which encodes a family of RNases thats act on the pistil and prevents the formation of the pollen tube when the S alleles presented in the pollen grain is the same as that is presented in the diploid tissue of the pistil. From the identification of the S alleles of plants of interest it is possible to guide the combinations between parents to obtain segregant populations and to predict the efficiency of apple tree genotypes when they are used as pollinators in commercial orchards. The objectives of this dissertation were to validate the use of DNA markers in the identification of the genetic constitution of the S locus of apple tree genotypes, as well as the definition of pollinators genotypes to be adopted in commercial orchards, serving as an auxiliary tool for the apple breeding. The study of the segregation of the S alleles carried out in segregating populations of apple trees by DNA markers (Chapter 2); the locus S of 28 genotypes of apple trees were analyzed by DNA markers and the genetic dissimilarity analysis was done based on the characterization of the same genotypes for based on the minimum descriptors required for the protection of new cultivars (Chapter 3); and the determination of the pollinators genotypes for three apple was done based on the genotyping of the S alleles associated with cross testing (Chapter 4). In the study of segregation of S alleles in segregating populations, the progeny of the cross between Fred Hough (S5S19) vs. Monalisa (S2S10) followed the expected segregation ratio for genotype compatibility: 1:1:1:1. In the other population evaluated, the same pair of S alleles, S3S5, were identified in both parents (M-11/01 and M-13/91), and the progeny showed the same genotype. In 26 of the 28 genotypes evaluated, both alleles S were identified, and in the remaining two genotypes only one allele was identified in each genotype. The average dissimilarity of the 28 genotypes obtained by the morphoagronomic characterization was 35 %. Considering the total genetic compatibility between the genotypes and the five major dissimilarities obtained, 15 crosses were suggested to increase the genetic base of the Epagri's Genetic Apple Breeding Program. Regarding the selection of pollinators, the field pollination tests did not show a significant difference between the cultivars and their respective pollinators tested for characters the fruit set and seed number, but when the S alleles present in each of the genotypes, were identified the presence of semi-compatibility cases between them. This fact can be explain by the high concentration of pollen grains applied on the pistil of the flowers on artificial pollination, which may mask the existing semi-compatibility. Considering the results obtained in this study, DNA markers can be used to identify the locus S genotype in apple trees as an auxiliary tool to the Epagri's Genetic Apple Breeding Program, both for the definition of combinations between parents for the formation of segregant populations as to chose pollinators to fruit-producing cultivars to be adopted in commercial orchards / Devido a autoincompatibilidade gametofítica (AIG), para a formação de frutos em plantas de macieira (Malus domestica Borkh.) é necessária a ocorrência de polinização cruzada entre plantas geneticamente compatíveis. A AIG é governada pelo loco multialélico S, que codifica para uma família de RNases atuantes no pistilo da planta e impede a formação do tubo polínico quando os alelos S presentes no grão de pólen forem iguais àqueles presentes no tecido diploide do pistilo. A partir da identificação dos alelos S de plantas de interesse é possível orientar as combinações entre genitores para a obtenção de populações segregantes via hibridações dirigidas e prever a eficiência de genótipos de macieira quando utilizados como polinizadores em pomares comerciais. Os objetivos dessa dissertação foram validar o uso de marcadores de DNA na identificação da constituição genética do loco S de genótipos de macieira e na definição de genótipos polinizadores a serem adotados em pomares comerciais, servindo como ferramenta auxiliar ao melhoramento genético de macieira. Realizou-se: o estudo da segregação dos alelos S em populações segregantes de macieira mediante genotipagem via marcadores de DNA (Capítulo 2); a genotipagem dos alelos S de 28 genótipos elite de macieira via marcadores de DNA e análise de dissimilaridade genética com base na caracterização quanto aos descritores mínimos requisitados para a proteção de novas cultivares (Capítulo 3); e a determinação dos genótipos polinizadores para três cultivares de macieira baseando-se na genotipagem dos alelos S associada à realização de cruzamentos teste a campo (Capítulo 4). No estudo da segregação dos alelos S em populações segregantes, a progênie do cruzamento entre Fred Hough (S5S19) vs. Monalisa (S2S10) apresentou a proporção de segregação esperada para compatibilidade entre genótipos: 1:1:1:1. Na segunda população avaliada, em ambos os genitores (M-11/01 e M-13/91) foi identificado o mesmo par de alelos S: S3S5, sendo que a progênie apresentou esse mesmo genótipo. Mediante a análise molecular, em 26 dos 28 genótipos avaliados foram identificados ambos os alelos do loco S, sendo que nos dois restantes apenas um alelo foi identificado em cada genótipo. A dissimilaridade média dos 28 genótipos identificada pela caracterização morfoagronômica foi de 35 %. Considerando a compatibilidade genética total entre os genótipos e as cinco maiores dissimilaridades obtidas, foram sugeridos 15 cruzamentos para ampliação da base genética do Programa de Melhoramento Genético da Epagri. Quanto a seleção de polinizadoras, os testes de polinização a campo não demonstraram diferença significativa entre as cultivares e suas respectivas polinizadoras para as características avaliadas, porém quando os alelos S foram identificados constatou-se a presença de casos de semicompatibilidade entre alguns dos genótipos avaliados. Sugere-se que a não identificação dos genótipos semicompatíveis ocorre devido a alta concentração de grãos de pólen aplicada sobre o pistilo das flores via polinização artificial das cultivares, que pode mascarar a semicompatibilidade existente. Considerando os resultados obtidos nesse estudo, a utilização de marcadores de DNA para identificação do genótipo do loco S em macieira pode ser empregada como ferramenta auxiliar ao programa de melhoramento genético de macieira da Epagri, tanto para a definição de combinações entre genitores para a formação de populações segregantes que serão alvo de seleção quanto para a escolha de polinizadoras geneticamente compatíveis com as cultivares produtoras de frutos a serem adotados em pomares comerciais, minimizando as perdas de produção em pomares comerciais devido a semicompatibilidade genética ou incompatibilidade entre os genótipos de macieira (cultivar copa x polinizadora)