The United States Fish and Wildlife Service (USFWS) expressed the need to develop appropriate management strategies for apparently high-density, growing black bear populations in the Roanoke-Neuse-Tar-Cape Fear ecosystem in coastal North Carolina and Virginia. In order to provide the scientific information necessary to develop these strategies, I investigated population densities and genetic structure of black bears at 3 national wildlife refuges [Great Dismal Swamp (GDSNWR), Pocosin Lakes (PLNWR), and Alligator River (ARNWR)]. Density estimates were derived from DNA samples collected noninvasively at each of the 3 refuges for 2 consecutive summers. Hair samples were analyzed for individual identification using 6-7 microsatellite markers. Estimated densities were some of the highest reported in the literature and ranged from 0.56-0.63 bears/km2 at GDSNWR to 0.65-1.12 bears/km2 at ARNWR to 1.23-1.66 bears/km2 at PLNWR. Sex ratios were male-biased in all areas of all refuges. Genetic variability and structure of bears at these refuges was assessed using 16 microsatellite markers for 40 bears from each refuge. Genetic variability of the 3 refuge populations was substantially high compared to other bear populations in North America, with observed heterozygosities ranging from 0.6729 at GDSNWR to 0.7219 at ARNWR. FST and DS values were relatively low (0.0257-0.0895 and 0.0971-0.3640, respectively), indicating movement of bears and gene flow across the landscape is adequate to prevent high levels of genetic differentiation and structure among the refuge bears. Genetic statistics at GDSNWR indicate that this population is isolated to some degree by geography (i.e., the Albemarle Sound) and encroaching urban development (i.e., the towns of Suffolk and Chesapeake). ARNWR has the potential to become isolated in the future if movement corridors to the south of the refuge are not maintained. Harvest of bears is likely warranted at PLNWR and ARNWR, though extreme caution must be taken the first few seasons as hunter success will be extremely high. Further research is needed to determine population growth rates, reproductive parameters, and survival rates at all 3 refuges, particularly if a hunting season will be established and maintained in these areas. Methods for regularly monitoring bear populations at these refuges also should be incorporated into biological programs, as bears comprise a significant component of the ecosystem at these refuges and cannot be ignored when outlining management goals. / Master of Science

Doctor of Philosophy / Department of Biology / Philip S. Gipson / Understanding factors that influence genetic variability, demographic vital rates, and resource selection is important for conservation and management of wildlife populations. I examined factors influencing microsatellite variability, demographic vital rates, and habitat use for a reintroduced elk (Cervus elaphus) population at Fort Riley, Kansas based on data collected from 2003 – 2007. Levels of allelic richness, observed heterozygosity, and expected heterozygosity for the Fort Riley population were intermediate to other North American elk populations. Genetic variability in restored North American elk populations was not well explained by founding population size, number of founding populations, or number of years since the last translocation. I examined the influence of demographic vital rates on the rate of population change to test the hypothesis that variability in calf survival has a greater influence on rates of population change than adult survival. Survival for prime-age adult elk had the highest stage-specific elasticity value, but life-stage simulation analysis indicated that variation in calf survival had the highest correlation with variation in population growth rate. These results suggest that calf survival varies temporally and is the vital rate most directly related to variation in population growth rate for this population. I assessed the relative influence of risk-related and resource-related factors on elk habitat selection by comparing predictor variables included in top resource selection function models at the landscape and home range scales. All predictor variables, with the exception of fall and spring prescribed burns, were included in top models across seasons at both spatial scales. Elk selected low elevation areas, gentle slopes, edge habitat, and areas close to streams at both spatial scales. At the landscape scale, elk generally avoided roads and preferred areas on or near Fort Riley. At both spatial scales, elk used riparian woodlands more frequently than grasslands and selected for agricultural crops when seasonally available. These findings do not support the idea that risk-related factors are the primary determinant of elk habitat use at the landscape scale as has been found for ungulates in areas with natural predators.

Thesis (PhD (Genetics))--University of Stellenbosch, 2011. / Includes bibliography / ENGLISH ABSTRACT: Grapevine diseases, in particular virus and virus-like diseases, are threatening grapevine industries worldwide; also in South Africa. Grapevine leafroll (GLR) is one of the most important diseases of grapevines, occurring in all grape-producing countries worldwide. Grapevine leafroll-associated virus 3 (GLRaV-3) is known to be closely associated with GLR disease and occurs commonly in South African vineyards. In this study three genetic variants of GLRaV-3 were identified in vineyards of the Western Cape, South Africaby single strand conformation polymorphism (SSCP) profiles generated from a region amplified in ORF5. A specific SSCP profile could be assigned to each variant group and these wereconfirmed by sequencing of the ORF5 regions.These results demonstrated that SSCP analysis on this region in ORF5 provides a fast and reliable indication of the GLRaV-3 variant status of a plant, which in many instances showed mixed infections. The full genome sequence of one representative of each variant group i.e. isolates 621 (group I), 623 (group II) and PL-20 (group III), was determined by sequencing overlapping cloned fragments of these isolates. The sequences of genomic 5’ ends of these isolates were determined by RLM-RACE. Sequence alignment of the 5’UTRs indicated significant sequence and length variation in this region, between the three South African variant groups. Nucleotide sequence alignment of the Hsp70h and CP gene regions of these isolates with those of isolates from elsewhere in the world, followed by phylogenetic analysis, further supported the presence of three GLRaV-3 variants in South Africa, and that two or three additional variant groups occurs elsewhere in the world. We further investigated the prevalence of these three GLRaV-3 variants in mother blocksof different cultivars and from different vine growing regions, using SSCP analysis. The majority of the plants studied, were infected with the group II variant, similar to isolates 623 and GP18. The distribution of the three GLRaV-3 variants within a spatio-temporally recorded cluster of diseased plants was studied by means of SSCP profile analysis. We showed that different GLRaV-3 variants are transmitted to adjacent plants in an infection cluster. Results showed that, in some leafroll disease clusters, the variant that was present in the original GLRaV-3 infected plant of a cluster was transmitted to adjacent plants in a row and across rows. Some plants in the cluster were also infected with variants not present in the original plant. These infections could have been caused by mealybug vectors feeding on plants from surrounding areas and then infecting these plants. The scientific information generated on GLRaV-3 variants in this project contributed to the advancement of our knowledge of genetic variability and provides a basis of further epidemiology and vector-virus studies. The study showed for the first time that different GLRaV-3 variants were transmitted to adjacent plants in a row and across rows in a GLR disease cluster. The diversity detected in the 5’UTR between variants from the three genetic groups provides a platform for the further study of the biological characteristics of GLRaV-3 variants. / AFRIKAANSE OPSOMMING: Wingerdsiektes, veral virus siektes, bedreig wingerd industrieë wêreldwyd, asook die Suid Afrikaanse wingerdbedryf. Rolbladsiekte is een van die belangrikste siektes op wingerd en kom wêreldwyd voor. Die virus, grapevine leafroll-associated virus 3 (GLRaV-3), word sterk geassosieer met Rolbladsiekte en kom wydverspreid voor in Suid Afrikaanse wingerde. Tydens hierdie studie is drie genetiese variante van GLRaV-3 geïdentifiseer in wingerd moederblokke in die Wes-Kaap. Die GLRaV-3 variante is geïdentifiseer met ‘n tegniek wat ‘single-strand conformation polymorphism (SSCP)’ genoem word. Die SSCP profiele was gegenereer vanaf PKR produkte van die ORF5 area op die genoom van GLRaV-3. Die geamplifiseerde produk van die ORF5 gebied is gebruik om die SSCP profiele te verkry en DNA-volgorde data in die gebied het die drie SSCP profiele gestaaf. Hierdie metode om virus variasie te bestudeer in plante is vinnig en betroubare resultate is verkry. Gemengde infeksies, wat gereeld in wingerd voorkom, kon ook met die tegniek opgespoor word. Die volledige nukleotied-volgorde van elkeen van die drie GLRaV-3 genome is volledig bepaal. Die isolate wat die drie variant groepe verteenwoordig is isolaat 621 (groep I), 623 (groep II) en PL-20 (groep III). Die nukleotiedvolgorde in die 5’UTR is bepaal met die RLM-RACE tegniek. Wanneer die 5’UTRs van die drie variante vergelyk is, het dit getoon dat daar verskille is in die volgordes en lengtes voorgekom het. Ander dele van die genoom, o.a. die dopproteïen (CP) en Hsp70 areas, is filogeneties vergelyk met isolate van regoor die wêreld. In die filogenetiese analise is bevind dat die drie GLRaV-3 variante saamgegroepeer het met ander isolate in die wêreld en dat daar elders ook twee to drie addisionele variant groepe van GLRaV-3 voorkom. Die verspreiding van die drie GLRaV-3 variante in wingerde is bestudeer in verskillende kultivars en in verskillende verbouingsgebiede. Die meerderheid van die plante in die studie was geïnfekteer met die groep II variant wat dieselfde is as isolate 623 en GP18. Die voorkoms van die drie variante in ‘n siekte cluster is bestudeer d.m.v SSCP. Die studie het gewys dat verskillende GLRaV-3 variante versprei word na aangrensende plante in ‘n ry en tussen rye. In sommige gevalle is die variant wat in die oorspronklik geïnfekteerde plant voorkom, oorgedra na naasliggende plante. Sommige van die plante in the infeksie area was ook met ander GLRaV-3 variante geïnfekteer wat moontlik deur wolluise oorgedra is vanaf naburige geïnfekteerde plante. Die wetenskaplike inligting wat tydens hierdie studie beskryf word aangaande die identifikasie van GLRaV-3 variante, dra by tot die molekulêre kennis van GLRaV-3 en verskaf ‘n basis vir verdure epidemiologiese -en insek oordragingstudies. Die studie het vir die eerste keer bewys dat verskillende GLRaV-3 variante na aanliggende plante in ‘n ry asook oor rye oorgedra word. Die diversiteit tussen die GLRaV-3 variant groepe in die 5’UTR moet verder ondersoek word en die deel van die genoom kan ‘n belangrike rol speel in die biologiese eienskappe van die variante.

Un inventaire des Tetrany chidae du Brésil a été réalisé dans 15 États ainsi que dans le District Fédéral, avec 550 échantillons de 120 espèces végétales collectés. Des infestations par des acariens tétranychidés ont été confirmées dans 207 de ces échantillons. Vingt-deux espèces appartenant à sept genres de la sous-familles des Bryobiinae et Tetranychinae ont été identifiées chez 58 hôtes différents. Trente-six nouvelles plantes hôtes pour 11 espèces ont été répertoriées au Brésil, en Amérique du sud ou dans le monde. De nouvelles localités ont été enregistrées au Brésil pour quatre tétranychides et une espèce a également été signalée pour la première fois en Amérique du sud. Quatre nouvelles espèces ont été découverte: deux appartenant au genre Oligonychus Berlese et deux appartenant aux genres Monoceronychus McGregor et Schizotetranychus Tragardh. La contribution à la systématique du genre Tetranychus a consisté à analyser conjointement des substuences d´ADN ribosomique (ITS) et mitochondrial (COI) de femelles de T.urticae, T. cinnabarinus (synonyme potentiel de T. urticae) et d'espèces très proches appartenant également au groupe Tetranychus sensu stricto. Cette étude a mis en évidence des incohérences ce qui nous a amené à remettre en question la fiabilité des données moléculaires concernant le groupe Tetranychus s. str. disponibles dans Genbank. Des données sur la variabilité génétique, la phylogénie et la structure de populations de T. urticae au Brésil et dans le monde sont ensuite présentées. Les résultats indiquent la présence significative d'une structuration génétique des populations de T. urticae par rapport à la localisation géographique. L´effet de la plante hôte n´est pas observé. La diversité haplotypique, inférée à partir des ces deux régions du génome (ITS et COI) est plus élevée dans les pays du pourtour méditerranéen. On a constaté la présence de deux haplotypes mitochondriaux (COI) au Brésil, l´un partagé avec la France, l´Espagne et les îles Canaries, et l´autre avec le Japon. / In this study we performed a survey of Tetranychidae mites from Brazil, including 15 States and the Federal District. A total of 550 samples of 120 different plant species were collected. Tetranychid mite infestations were confirmed in 207 samples, and 22 species belonging to seven genera of the Bryobiinae and Tetranychinae subfamilies were identified on 58 different host plants. Thirty-six new hosts were recorded in Brazil, South America and worldwide for eleven species. New localities were registered for four tetranychid genera and a new record to South America was confirmed. Four species were identified as new for science: two belonging to the Oligonychus Berlese genus and two belonging to the Monoceronychus McGregor and Schizotetranychus Tragardh genera. We analyzed and evaluated the identity of 105 Genbank accessions of ITS2 rDNA and 138 COI mtDNA sequences which were deposited as T. urticae and as fourteen other taxa morphologically close ly related to Tetranychus sensu stricto. Among the deposited sequences in the Genbank, numerous cases of apparently mistaken identities were identified in the group Tetranychus s. str., especially between T. urticae, T. cinnabarinus, T. kanzawai and T. truncatus. New sequences of ITS and COI were obtained from individuals collected in Brazil and some localities of the Palearctic Region (France, continental Spain, Canary Islands, Greece, Syria, Tunisia, Poland and Norway plus one from Canada). While significant differences were detected on population genetic structure of the analyzed samples according to the geographic region, any effect of the host plant was observed. Haplotype diversity inferred from both ITS and COI sequences was higher in samples from the Mediterranean basin. ITS sequences obtained from Brazil samples were homogenous, and two COI haplotypes were found, one of them also present in France, Spain and the Canary Islands and the other in Japan.

L'artichaut et le cardon, appartenant à la famille des Asteraceae (Compositae), sont des plantes pérennes herbacées natives du bassin méditerranéen, et qui sont traditionnellement cultivées comme plantes maraîchères, respectivement pour leurs têtes et leurs cardes. L'Italie est le pays possédant la plus importante collection de germoplasmes autochtones d'artichaut. Dans le centre de l'Italie, le type Romanesco est étendu. Ces dernières années, le développement des techniques in vitro a permis la multiplication de l'artichaut Romanesco et sa rapide expansion. Le clone Romanesco C3 a ainsi remplacé de nombreuses races locales de Romanesco, contribuant de la sorte à une érosion significative des ressources génétiques locales. Concernant le germoplasme de cardon cultivé, seules quelques études sont disponibles sur son identification et sa caractérisation génétique. Le cardon sauvage, n'est pas du tout cultivé; il est davantage considéré comme une adventice dans le paysage italien. La grande variabilité existant entre les espèces de Cynara n'est pas correctement décrite et il existe plusieurs cas d'homonymes. Le secteur de l'artichaut italien a du faire face ces dernières années à une crise causée principalement par l'apparition sur le marché de produits étrangers et par les coûts élevés de mise en culture et de récolte. Afin de surmonter cette crise, de nombreuses valorisations du Cynara ont été envisagées. Ces potentielles applications pour la culture ont pu voir le jour grâce au support de l'Union européenne pour la recherche sur les co-poduits issus de l'agriculture, et ont mené à un intérêt croissant pour la biomasse entière d'artichaut. Considérant ces remarques préliminaires, une stratégie, qui permettrait de valoriser le germoplasme italien par la production concomitante de biomasse et de biocomposés, a été mise en place durant ces trois ans de doctorat. Le premier objectif de ce travail de doctorat consistait en i) la caractérisation agro-morphologique du germoplasme italien par le biais de descripteurs UPOV, ii) l'évaluation de la variabilité génétique à l'intérieur et entre les races/clones et iii) l'identification et la préservation des ressources génétiques pour le développement de futurs programmes d'amélioration des plantes. Suite à cette caractérisation, trois génotypes ont été sélectionnés et enregistrés sous les noms de Michelangelo, Donatello and Raffaello. Afin d'analyser le germoplasme italien de Cynara d'un point de vue de la biomasse, différents traits expliquant la vigueur de la plante et la production de matière sèche ont été considérés. Le rendement en biomasse aérienne s'est révélé très élevé, soulignant la possibilité d'utiliser cette culture comme matière première industrielle. Un point particulier du programme de thèse était de mettre au point les méthodes d'extraction de biocomposés et les techniques d'analyse afin d'optimiser le rendement en polyphénols à partir de la biomasse de Cynara à l'échelle laboratoire. L'ASE a été reconnue comme étant la meilleure technique. De plus, les cinétiques de production de biomasse et de biocomposés ont été évaluées et le stade physiologique optimal pour collecter le matériel végétal en champ a été identifié. La caractérisation biochimique a été réalisée grâce aux méthodes mises au point et en collectant le matériel végétal au stade physiologique optimal identifié afin de distinguer les génotypes les plus appropriés pour la production de biocomposés. Le dernier point du programme de thèse était centré sur le développement d'une technique alternative de production de biomasse et de biocomposés en conditions de croissance sous serre. Les résultats obtenus mettent en exergue la possibilité d'utiliser avec succès certains génotypes de Cynara pour la production de biomasse et de biocomposés. La perspective réelle d'utiliser certains génotypes d'artichaut pour une double valorisation alimentaire et non-alimentaire a ainsi été soulignée. / Globe artichoke and cardoon, belonging to the Asteraceae (Compositae) family, are herbaceous perennial plants native to the Mediterranean area, which are traditionally grown as vegetables for the heads and the fleshy petiole leaves, respectively. Italy is the richest reserve of globe artichoke autochthonous germplasm, which is vegetatively propagated and well adapted to the different pedoclimatic conditions of the Country. In Central Italian environments, the Romanesco type is widespread. In the last years, the development of in vitro technologies allowed the propagation of Romanesco globe artichoke type and its rapid expansion. As a result, the micropropagated Romanesco clone C3 has replaced many landraces traditionally grown in the Latium Region and has led either to a significant erosion of local genetic resources. As regards Italian cultivated cardoon germplasm, there are few studies on its genetic characterization and identification and there is a lack of information about the genetic variability existing within and among accessions. For the wild cardoon, no specialized crop is present and it represents mainly a weed in Italian environments. The great variability existing in Cynara spp. has not been described, the nomenclature of Italian germplasm is not always very clear since there are many cases of homonyms. In addition, Italian globe artichoke sector is facing a crisis due principally to the appearance on the market of foreign products and to the high labor cost required for crop cultivation and harvesting. In order to overcome this crisis several possible industrial uses were considered for the species. Considering these preliminary remarks, a strategy for valorizing Italian germplasm using biomass and biocompound production has been carried out during the three years of PhD program. The first objective of PhD work consisted in (i) characterizing agro-morphologically Italian germplasm using UPOV descriptors, (ii) assessing the genetic variability existing within and among landraces/clones and (iii) identifying and preserving genetic resources for the development of future plant breeding programs. As a result of this characterization, three genotypes have been selected and registered under the names of Michelangelo, Donatello and Raffaello. In order to analyze Italian Cynara spp. germplasm also from a biomass point of view, different traits explaining plant vigor and dry matter production have been considered. The aerial biomass yield resulted very high underlining the possibility of using this crop as raw industrial material. A focal point of PhD program was to set up biocompound extraction methods and analysis techniques to optimize polyphenol recovery from biomass of Cynara spp at a laboratory scale. In particular, ASE was found as the best extraction technique which allows to reduce extraction time and solvent consumption, increase nutraceutical yield and improvement of extract quality. Moreover, the kinetics of biomass and bio-compound production has been evaluated and the optimal physiological stage to collect plant material grown in open field has been identified. Biochemical characterization has been performed using the methods set up and collecting plant material in the optimal physiological stage identified in order to distinguish which genotypes were more suitable for bio-compound production purpose. The last focal point of PhD program was the development of an alternative technique for biomass and biocompound production in greenhouse grown conditions. Results obtained in the three PhD years, highlighted the possibility of using successfully some Cynara spp. genotypes for biomass and bio-compound production, in particular in open field condition. Also the real prospect of using some globe artichoke genotypes for food and non-food dual-production (biomass for biocompound extraction and heads for human food) has been underlined.

A tolerância ao Cádmio (Cd) é um assunto de relevada importância, devido aos vários problemas que este metal pode causar para a agricultura, levando à queda de produção e perda da qualidade dos alimentos, representando, também, riscos à saúde humana pelo consumo de produtos contaminados com esse metal. Neste trabalho, fizemos um estudo geral sobre a tolerância ao Cd em plantas de tomateiro, partindo de 3 abordagens complementares, onde geramos variabilidade genética por meio de mutagênese usando um tomateiro modelo (cultivar Micro-Tom); exploramos uma pequena fração da variabilidade genética da espécie para identificar genótipos com diferentes graus de tolerância ao Cd e, finalmente, realizamos estudos ligados ao metabolismo oxidativo de duas cultivares selecionadas, sendo uma sensível e outra mais tolerante a este metal. Este trabalho nos conduziu também ao desenvolvimento e adaptação de metodologias para a avaliação e seleção de plantas tolerantes a metais pesados, dentre as quais propomos a utilização de um Índice de Tolerância adequado para avaliar cultivares morfologicamente diferentes. Por fim, os resultados obtidos neste trabalho possibilitaram uma visão geral sobre os parâmetros de tolerância ao Cd em plantas de tomateiro, bem como o estudo dos principais padrões de resposta no metabolismo oxidativo de genótipos mais sensíveis e mais tolerantes a este metal. / Tolerance to heavy metal Cadmium (Cd) is an important subject because this metal can cause several problems in agriculture, such as decrease in production and loss of food quality, representing risks to human health by consumption of vegetables contaminated with this metal. In this research, we studied Cd-tolerance in tomato plants, using three complementary approaches, generating genetic variability by mutagenesis using a tomato model (cultivar Micro-Tom), exploring a small fraction of the genetic variability of species to identify genotypes with different degrees of Cd-tolerance and, finally, we conducted studies related to the oxidative metabolism of two cultivars, with low and high tolerance to this metal. Additionally, this work results in the development and adaptation of methodologies for the evaluation and selection of tolerant plants to heavy metals. Then, we propose an appropriate Tolerance Index to evaluate morphologically different cultivars. In conclusion, the results of this study are an overview of the parameters of Cd-tolerance in tomato plants, as well as the study of the some patterns of response in the oxidative metabolism of genotypes more sensitive and more tolerant to this metal.

O gênero Dioscorea é o mais amplo da família Dioscoreaceae, apresentando cerca de 600 espécies distribuídas, sobretudo, nos trópicos, com grande importância na alimentação, principalmente na África Ocidental, na Sudeste Asiático, no Caríbe e em alguns países da América do Sul. No Brasil, algumas espécies de inhame (Dioscorea spp.), juntamente com a mandioca (Manihot esculenta Crantz), têm uma profunda importância na agricultura de subsistência, sendo utilizadas basicamente como fonte de carboidrato para alimentação humana. Pouco se conhece sobre a diversidade e estrutura genética dessas espécies, como evoluíram nos últimos anos, sobretudo pela escassez de avaliações moleculares. O presente estudo tem como objetivos: (i) apresentar dados sócio-econômicos e etnobotânicos relativos aos diferentes agricultores que cultivam a espécie; (ii) isolar primers de microssatélites usando uma biblioteca genômica enriquecida e testar sua amplificação entre outras espécies de Dioscorea; (iii) analisar a relação genética entre 73 variedades locais e 17 acessos comerciais de inhame coletados em cinco diferentes regiões do Brasil (Sul, Sudeste, Nordeste, Norte e Centro-oeste) usando um conjunto de 12 microssatélites. Túberas de inhame foram coletadas em 28 municípios proveniente de cinco regiões onde a espécie é comumente cultivada, bem como em mercados locais e feiras de vários estados do Brasil. Outros acessos foram obtidos dos bancos de germoplasma ex situ pertencentes a ESALQ/USP, IAC e FCA/UNESP. Uma análise descritiva de diferentes agricultores foi realizada, indicando distintos perfis entre as regiões analisadas. Um amplo espectro de nomes populares foi registrado com diferenças entre regiões. As entrevistas com os agricultores revelaram que a espécie tem perdido sua importância em algumas áreas tradicionais/locais. O isolamento de marcadores microssatélites polimórficos resultou na detecção de 14 locos de microssatélites (SSR). Destes, dez foram selecionados para caracterizar 80 acessos de D. alata. O conteúdo de informação polimórfica foi de 0,39 a 0,78 e o poder de descriminação foi de 0,15 a 0,91. Seis destes marcadores mostraram transferibilidade entre as espécies D. bulbifera, D. cayenensis-D. rotundata e D. trifida. Em um estudo de diversidade morfológica e molecular, 12 pares de microssatélites polimórficos (nove desenvolvidos neste estudo e três obtidos da literatura) foram usados para gerar perfis de DNA de cada acesso da espécie e quatro perfis morfológicos foram analisados. A caracterização morfológica mostrou considerável diversidade e nenhum agrupamento específico foi observado entre as regiões. As análises moleculares de D. alata mostraram alta diversidade intra-específica nos acessos locais de diferentes regiões do Brasil. Contudo, a estrutura populacional entre as regiões coletadas foi relativamente baixa. Somente acessos da região Centro-Oeste apresentaram um aparente agrupamento regional, resultado quase similar foi observado nos acessos do nordeste. Estes resultados mostram uma mistura de acessos em todas as regiões coletadas, na qual é consistente com a falta de correlação entre as distâncias geográficas e genéticas, sugerindo que as túberas de inhame têm se movido extensivamente pelo fluxo humano. A diversidade genética encontrada pode ser explicada pelo resultado de um contínuo intercâmbio de variedades através do território brasileiro. O desenvolvimento de marcadores moleculares SSR em D. alata e análises de genética populacional são essenciais para o avanço de pesquisas nesta espécie. A geração de informação é de grande importância para a identificação, exploração racional e conservação da variabilidade genética da espécie, tanto da forma in situ e/ou ex situ. / The genus Dioscorea is the largest in the Dioscoreaceae family, featuring approximately 600 species distributed mainly in the tropics, with high importance as food supply in West Africa, Asia southeast, the Caribbean and a few countries in South America. In Brazil, some species of yam (Dioscorea spp.) together with cassava (Manihot esculenta Crantz), have a profound importance in subsistence agriculture, being used primarily as a source of carbohydrate for human feeding. Little is known about the genetic diversity and structure of these species, and also how it evolved in recent centuries, particularly because of the scarcity of molecular evaluations. The present study is intended to: (i) present socio-economic and ethnobotanical data on the different agriculturists that cultivated the species; (ii) isolate microsatellite primers using an enriched genomic library technique and test for cross amplifications in other species of Dioscorea; (iii) analyse the genetic relationships among 73 local acessions and 17 commercial accessions of water yam collected in five different regions in Brazil (South, Southeast, Northeast, Central-West and North) using a set of 12 microsatellite. Tubers of D. alata were collected in 28 municipalities from this five regioesn where the species is commonly cultivated, as well as on local markets and fairs from several states across Brazil. Other accessions were obtained from the ex situ germplasm collections belonging to ESALQ/USP, IAC and FCA/UNESP. A descriptive analysis of different farmers was performed, indicating unequal profiles between the screened regions. A wide range of vernacular names were registered with differences between regions. The interviews eith the agriculturist revealed that the species are losing their importance in some traditional/local areas. The isolation of codominant polymorphic microsatellite markers resulted in the detection of 14 short tandem repeat (SSR) loci, and ten were selected to characterize 80 D. alata accessions. The polymorphism information content varied from 0.39 to 0.78 and the power discrimination ranged from 0.15 to 0.91. Six of the markers showed transferability between the species D. bulbifera, D. cayenensis-D. rotundata and D. trifida. In a morphological and molecular diversity study, 12 polymorphic microsatellite primers were used to generate DNA profiles for each accession of the species and four morphological traits were analyzed. The morphological characterization showed considerable diversity and no specific clustering was observed between regions. The molecular analyses of D. alata showed a high intraspecific diversity in local varieties from different regions in Brazil. However, population structuring between sampling regions was rather low. Only the accessions from the Central-Western region showed an apparent regional clustering, almoust similar were observed with northeast acessions. These results show an admixture of accessions in all sampling regions, which is further consistent with the lack of a correlation between geographic and genetic distances, suggesting that water yam tubers have moved extensively by human fluxes. The genetic diversity found can be explained by the result of a continuous exchange of varieties through the Brazilian distribution range. The development of molecular SSR markers for D. alata and genetic population analysis is essential for the ongoing research on this species. The generated information is of great importance for the identification, rational exploitation and conservation of the genetic variability of this species, in a in situ and/or ex situ way.

Made available in DSpace on 2016-08-10T10:55:06Z (GMT). No. of bitstreams: 1 Nicole Cristina Lopes Dutra.pdf: 1587085 bytes, checksum: 7c8f39f69da6456b027da52bf00a0794 (MD5) Previous issue date: 2006-11-29 / Studies that concern Cerrado (Brazilian s Savannah) fauna are important as we consider the elevated rate of destruction of its landscapes in the last decades. Bothrops moojeni is one of the species of vipers that occur on the Cerrado area and its venom has shown great applicability in health. In the last years, molecular markers have helped in researches that aim to assess the genetic variability. In the present work, we perform the characterization of 5 populations of Caiçaca (Bothrops moojeni) using RAPD (Random Amplified Polimorphyc DNA) markers and also used a scienciometric approach to highlight the importance of this species in health in the site Institute for Scientific Information (ISI), using the method General Search with the key words: serpente , Bothrops and Bothrops moojeni . The obtained Data were used to estimate the magnitude and distribution of variability within and among the groups using AMOVA. Aiming to analyze the patterns of spatial variation patterns, Pearson relation coefficient (r) was estimated between the matrixes of genetic distances and geographic among the populations. With the 5 primers were obtained 59 loci, from which 81 porcent (48) were polymorphic, ranging between 35% and 69% in the groups. The value of f ST obtained with AMOVA was equal to 0,13 (P smaller 0,0001 - 10000 permutations). Thus, it was possible to observe that Caiçaca populations studied presented a low level of genetic variability (He = 0,19) and most of the genetic variation is within the populations, what might indicate that is already occurring a reduction of the rate of gene flow between the populations. Considering the geographic area studied (lager distance was equal to 851,83 km), populations that are geographically near presented a high level of genetic diversity and the genetic variability is not spatially structured. The populations of Bothrops moojeni studied presented low levels of Genetic diversity for the RAPD loci evaluated. Genetic variability is not spatially structured. Concerning scienciometry analyses, the following key-words: serpente , Bothrops and Bothrops moojeni most of the studies concerning zoology (51 porcent), Pharmacology and Pharmacy (32 porcent) and Biochemistry and Molecular Biology (23 porcent), there was no paper published till the present date, that aimed to evaluate genetic variability in this species, as well as in the genus Bothrops. We recommend a wide-ranging study to evaluate genetic variability in this species due to its importance to conservation of the ecosystem from which it belong as well due to the great interest in health. / Estudos sobre a fauna do Cerrado são importantes considerando a alta taxa de destruição das suas paisagens naturais durante as últimas décadas. Estudos com marcadores moleculares se fazem importantes, pois auxiliam nas pesquisas que pretendem acessar a variabilidade genética para fins de conservação. Entre as espécies do Cerrado, Bothrops moojeni é uma serpente que vem demonstrando grande aplicabilidade na saúde. Neste sentido, o objetivo deste trabalho foi caracterizar a variabilidade genética de cinco populações de Caiçaca (Bothrops moojeni Hoge, 1966) a partir de marcadores RAPD (Random Amplified Polimorphyc DNA), bem como realizar um levantamento cienciométrico da importância da espécie para a área da saúde no sítio Institute for Scientific Information (ISI), utilizando o método GeneralSearch com a palavras chaves: serpentes, Bothrops e Bothrops moojeni. Os dados gerados com marcadores RAPD foram utilizados para avaliar a magnitude e a distribuição da variabilidade entre e dentro das populações por meio da Análise de Variância Molecular (AMOVA). A fim de se avaliar os padrões de variação espacial, foi estimado o coeficiente de correlação de Pearson (r) entre a matriz de distâncias genéticas e geográficas entre as populações. Com base nos cinco primers foram obtidos 59 locos, dos quais 81 por cento (48) foram polimórficos, variando entre 35 por cento e 69 por cento nas populações. O valor do Fí ST obtido pela AMOVA foi igual a 0,13 (P menor 0,0001 - 10000 permutações). Foi possível observar que as populações de Caiçaca estudadas apresentam baixos níveis de variabilidade genética (He = 0,19) para os locos RAPD estudados, sendo que a maior parte da variação encontra-se dentro das populações, o que poderia indicar que já está ocorrendo uma redução na taxa de fluxo gênico entre as populações. Considerando a escala geográfica em estudo (maior distância igual a 851,83 km), populações que estão próximas geograficamente apresentam uma considerável divergência genética e a variabilidade genética não se encontra estruturada espacialmente. No que diz respeito a cienciometria, nas palavras-chave serpente , Bothrops e Bothrops moojeni a maioria dos estudos foi sobre zoologia (51 por cento), Farmacologia e Farmácia (32 por cento) e Bioquímica e Biologia Molecular (23 por cento), não existindo nenhum trabalho publicado, até a presente data, que avalie a variabilidade genética desta espécie. Deve-se proceder com uma análise mais sistemática e de maior magnitude para avaliar a variabilidade genética desta espécie que além de sua importância para a conservação é uma espécie de muito interesse para a área de saúde.

O Fusarium spp. é um fungo cosmopolita, compreendendo uma grande quantidade de espécies que são conhecidas por causar doenças em culturas de importância agronômica. Embora, isolados não patogênicos de Fusarium spp. tenham sido descritos, pouco se conhece sobre a variabilidade genética deste grupo, ainda que estejam presentes em inúmeros locais. Assim sendo, este trabalho teve como objetivos ampliar estes conhecimentos, avaliando a variabilidade genética e forma de interação de isolados patogênicos e não patogênicos de Fusarium spp. obtidos de diferentes hospedeiros. Desta forma, 83 isolados de Fusarium spp. foram avaliados por meio das técnicas de ARDRA, sequenciamento do rDNA e RAPD. A análise por meio de ARDRA, permitiu a distinção dos 83 isolados de Fusarium spp. em 19 haplótipos, apresentando uma grande diversidade dentro de cada haplótipo, mas de forma geral os isolados patogênicos e não patogênicos de Fusarium spp. não puderam ser discriminados. Nas análises de sequenciamento da região ITS do rDNA, foi observado que isolados de Fusarium se agruparam independentemente da espécie. Estes resultados comprovam a necessidade de uma revisão taxonômica dentro do gênero Fusarium. A análises por marcadores RAPD revelou que os 83 isolados de Fusarium spp. avaliados neste estudo apresentaram ampla variabilidade a qual não está correlacionada com a característica taxonômica. Entretanto, foi observado que isolados patogênicos e endofíticos de F. oxysporum obtidos de soja são geneticamente diferentes. Quanto às análises de interação de isolados patogênicos e não patogênicos de Fusarium spp. com cultivares susceptíveis de tomate e soja, verificou-se que estes isolados interagiram de forma diversa, nos diferentes tecidos vegetais, sendo que alguns isolados endofíticos promoveram o crescimento vegetal. Dentre estes, destaca-se o isolado endofítico Cac19.4 que mostrou-se geneticamente diferente de isolados patogênicos de Fusarium spp., além de promover um aumento de peso da raiz e caule de plântulas de soja e tomate. Dessa forma este isolado poderia ser selecionado para futuras análises, visando um melhor aproveitamento deste microrganismo endofítico em estudos de interesse agronômico. / Fusarium spp. is a cosmopolitan fungus that covers a great number of species known by the ability of causing diseases in agricultural important crops. Although non-pathogenic isolates of Fusarium spp. have been described, little is known about the genetic variability of this group, even if it can be found in countless places. Therefore, this work had the objectives of increase this knowledge, evaluating the genetic variability and modes of interaction between Fusarium spp pathogenic and non-pathogenic isolates, obtained from different hosts. In this way, ARDRA, rDNA sequencing, and RAPD techniques evaluated 83 Fusarium spp. isolates. The ARDRA analysis allowed the separation of 83 isolates in 19 haplotypes. In spite of the great diversity found inside each haplotypes, in general it was difficult to distinguish pathogenic and nonpathogenic isolates. In the sequencing analysis of the ITS region of rDNA, it was observed that Fusarium isolates were grouped together independently to the species. These results proved the need for a taxonomic review inside Fusarium genera. The RAPD analysis revealed that the 83 Fusarium spp. isolates evaluated in this study presents high levels of variability not correlated with the taxonomic characteristic. However, we observed that pathogenic and endophytical F. oxysporum isolated from soybean are genetically different. The interaction analysis between pathogenic and non-pathogenic isolates of Fusarium spp. with susceptible cultivars of tomato and soybean, showed different modes of interaction on different plant tissues, where some endophytical isolates increased plant growth. Among these, we can emphasize the endophytic isolate Cac19.4, that is genetically different if compared with Fusarium spp. pathogenic isolates, in spite of his ability to promote an increase in root and stems weight of soybean and tomato plantlets. Thus, this isolate could be selected for further analysis, looking for a better use of this endophytical microorganism in agricultural interest studies.

Os objetivos do presente trabalho se dirigem ao estudo da variabilidade genética e da depressão por endogamia em sete populações de milho de ampla base genética, visando ao melhoramento de populações e obtenção de linhagens endogâmicas promissoras. Foram instalados onze experimentos em blocos casualizados em um local (Anhembi, SP), com diferentes conjuntos (N) de progênies S1 obtidos de sete populações (GO-D: dentado, GO-F: flint, GO-L: espiga longa, GO-G: espiga grossa; e compostos G3, G4 e GO-S). Foram estimadas a variância genética entre médias de progênies (2G), a variância fenotípica entre médias de progênies?^ (2F?^) e o coeficiente de herdabilidade (sentido amplo) para médias de progênies (2X). As estimativas de h 2Xhforam altas para peso de espigas (PE: 0,89 a 0,94), comprimento da espiga (CE: 0,77 a 0,88) e diâmtero da espiga (DE: 0,77 a 0,92); e menores para altura da planta (AP: 0,58 a 0,80) e altura da espiga (AE: 0,54 a 0,84), demonstrando alto potencial das populações para seleção recorrente com progênies S1. A variável PE nas populações base usadas como testemunha, mostrou valores variando de 11200 kg.ha-1 (GO-D) a 12800 kg.ha-1 (G3). As médias de progênies S1 entre populações variaram de 6070 kg.ha-1 (GO-F) a 7380 kg.ha-1 (G4); a depressão por endogamia nas progênies S1 variou de 37,5% (G4) a 48,0% (G3) em relação à população base. Os estudos sobre endogamia envolvendo as sete populações foram conduzidos com amostras da população original não endógama (S0) e das gerações S1 e S2 de autofecundação. Os experimentos foram conduzidos em Londrina (PR) e Piracicaba (SP) em blocos casualizados com parcelas subdivididas, com as populações representadas nas parcelas e as gerações de endogamia nas sub-parcelas. A estimação da depressão por endogamia foi obtida pelo modelo de regressão linear Y = µ0 + ?, sendo ? a depressão por endogamia para 100% de homozigose. A depressão esperada para 50% de homozigose é ?/2, cujo valor em percentagem variou de 25,4% a 41,4% em Piracicaba e de 23,1% a 39,3% em Londrina. Para os demais caracteres, os efeitos depressivos foram menores, geralmente <25% para AP e AE e <15% para DE e CE. / The objectives of the present work were directed for the study of genetic variability and inbreeding depression in seven maize populations of broad genetic base, as a guide for population improvement and development of promising inbred lines. The field evaluation was in eleven experiments (randomized complete blocks) in one location (Anhembi, SP) with different groups (N) of S1 progenies obtained of seven populations (GO-D: dent type, GO-F: flint type, GO-L: long ear, GO-G: thick ear; and composites G3, G4 e GO-S). Estimates were obtained for genetic variance (?^: progeny mean basis), phenotypic variance of progeny means (2G2F?^), and coefficient of heritability (broad sense) for progeny means (2Xh). Estimates of 2Xhwere high for ear weight (PE: 0.89 to 0.94), ear length (CE: 0.77 to 0.88) and ear diameter (DE: 0.77 to 0.92); and lower for plant height (AP: 0.58 to 0.80) and ear height (AE: 0.54 to 0.84), thus showing the high potential of the populations for recurrent selection based on S1 progenies. Ear yield (PE) in the base populations used as ckecks varied from 11200 kg.ha-1 (GO-D) to 12800 kg.ha-1 (G3). The means of S1 progenies varied from 6070 kg.ha-1 (GO-F) to 7380 kg.ha-1 (G4); the inbreeding depression in S1 progenies varied from 37.5% (G4) to 48.0% (G3) relative to the non-inbred population. For the studies on inbreeding in the seven populations samples of the original non-inbred populations (S0) and S1 and S2 generations of inbreeding were used. Filed experiments were carried out in Londrina (PR) and Piracicaba (SP) in randomized blocks with spli-plots, where populations were in the whole plots and inbreeding generations in the sub-plots. The estimates of inbreeding depression were obtained by the linear regression model Y = µ0 + ?, where ? is the iinbreeding depression for 100% homozygosity. The expected inbreeding depression for 50% homozygosity is ?/2, and the estimates in percentage varied from 25.4% to 41.4% in Piracicaba and from 23.1% to 39.3% in Londrina. For the other traits the inbreeding effects were lower, in general <25% for AP and AE and <15% for DE and CE.