The aim of this study was to obtain the molecular marker of Taiwania cryptomerioides Hayata based on DNA sequence data of PCR- sequencing and inter-simple sequence repeat (ISSR), and to evaluate the genetic diversity of populations of Taiwania cryptomerioides Hayata and molecular phylogeny of T. cryptomerioides and Taiwania flousiana Gaussen. The sequence data based on the internal transcribed spacer (ITS) of a total of 108 samples of T. cryptomerioides were determined. Eight different populations of T. cryptomerioides and 12 samples of T. flousiana from Yunnan, China were analyzed. The finding of the study showed that heterogeneity of ITS region within individuals of T. cryptomerioides was high by showing high nucleotide diversity among ITS sequences both in T. cryptomerioides ( £k = 0.18153) and T. flousiana ( £k = 0.19751). The findings fit in Tajima¡¦s D test of neutrality based on DNA sequence variation in the ITS region of T. cryptomerioides and T. flousiana. It is not obvious to incorporate into different population through clustering analysis based on data of the ITS region of T. cryptomerioides and T. flousiana. However, slightly genetic differentiation between T. cryptomerioides and T. flousiana was found, which figured of Fst (Fst = 0.0441~ 0.0856, an average value = 0.0611). On the other hand, the samples were studied by using ISSR markers. Of the 100 primers screened, 4 produced highly reproducible ISSR bands, and 24 discernible DNA fragments were generated with 17 being polymorphic. Based on cluster analysis of molecular data, the cluster is not clear among populations of T. cryptomerioides and T. flousiana. The analysis of AMOVA revealed that the variance component between species of T. cryptomerioides and T. flousiana was 38.54¢H (P < 0.001); however, the variance component within species is 61.46 (P < 0.001). The variation within population of T. cryptomerioides was 84.74¢H (P < 0.001) and the variance between populations is 15.26¢H (P < 0.001), indicating that the genetic diversity of individuals within population was high. The aforementioned data suggest that gene flow among different populations of T. cryptomerioides was high, indicating that the genetic diversity was high among individuals of T. cryptomerioides but was low between populations. Furthermore, it is concluded both species are genetically closer and could be grouped into the same species.

The objective of this study is to elucidate the efficiency of enation-structure (at node) recognition method at pre-flowing stage and to understand the population genetic variation of the Mikania weeds in Taiwan. The plant materials collected by recognizing enation-structure symptom method from North, Central, South and East Taiwan and off-shore islands. Using PCR¡V sequencing marker techniques, the sequencing revealed that nrDNA ITS region could identify three Mikania weeds and the 97% similarity of phylogenetic relationship between M.cordata and M.micrantha are more closer than that between M.cordata and M.scandens, whose ITS sequence is obtained from GeneBank. However ,the sequences of chloroplast DNA of M.cordata and M.micrantha at trnL intron¡]436bp¡^or trnL-trnF IGS¡]345 bp¡^are almost the same and could not be used as molecular markers. The recognizing techniques of enation-structure was supported by the nrDNA ITS region and ISSR results at the end, thus the finding can be recommended to the Council of Agriculture in order to eliminate the weed and to reduce the impact on M.cordata, which is native in Taiwan. Moreover, the findings of ISSR analysis in the aspect of population genetic variation indicated that high genetic differentiation ¡]Gst>0.5¡^was found among the M. cordata and M.micrantha populations. Based on the Mantle test, there was no relationship between genetic distance and geographic distance in M.micrantha¡]r=0.0053,p=0.47¡^.This phenomenon revealed that populations of M.micrantha had complex population variability within the short-term invaded into Taiwan that might be resulted from the random dispersion of human activities. The population of M.micrantha was established by few individuals (founders) and grown rapidly in Taiwan, resulting in population differentiation via genetic drift. In contrast to M.micrantha ,there was relationship between genetic distance and geographic distance in M.cordata ¡]r=0.44,p=0.025¡¯¡^.It revealed that populations of M.cordata agreed to the concept of isolation by distance model, which might be evolved from the result of natural dispersion. In conclusion, the population structure of M.micrantha in Taiwan is stable , suggesting that control of Mikania population should be based on different populations where have large differentiation among them .

Submitted by Amanda Silva (amanda.osilva2@ufpe.br) on 2015-04-08T12:53:20Z No. of bitstreams: 2 Dissertação Nathália Leite.pdf: 1163803 bytes, checksum: ed3fcb6e6b34f0c3e3452518a29c3263 (MD5) license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) / Made available in DSpace on 2015-04-08T12:53:20Z (GMT). No. of bitstreams: 2 Dissertação Nathália Leite.pdf: 1163803 bytes, checksum: ed3fcb6e6b34f0c3e3452518a29c3263 (MD5) license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Previous issue date: 2012-02-29 / CNPq / O presente estudo foi realizado com os objetivos de identificar acessos de feijão-caupi tolerantes ao caruncho (C. maculatus) e caracterizar esses acessos a nível molecular, em conjunto a acessos de V. unguiculata ssp. cylindrica e V. radiata, por meio de marcadores DAF (DNA Amplification Fingerprinting) e ISSR (Inter Simple Sequence Repeat), visando identificar parentais promissores para futuros trabalhos de melhoramento, bem como para mapeamento, pela associação com o nível de resposta ao caruncho. Para identificação da tolerância ao caruncho em feijão-caupi, 27 acessos foram avaliados em ensaios de laboratório, utilizando-se o Delineamento Experimental Inteiramente Casualizado. Para cada acesso, uma amostra com 30 grãos de feijão-caupi foi infestada com cinco casais de caruncho com até 24 h de idade, sendo os resultados obtidos submetidos à análise estatística e suas médias comparadas pelo teste de Tukey a 5% de probabilidade. Para as duas variáveis estudadas, o acesso INHUMA comportou-se como suscetível, enquanto os acessos PATATIVA e MNC99-537F-4 apresentaram-se tolerantes para ambas as variáveis. Os demais acessos testados apresentaram comportamento intermediário, o que não os diferencia em relação à INHUMA, PATATIVA e MNC99-537F-4. Na caracterização molecular dos acessos, 25 primers geraram um total de 239 amplicons, dos quais 163 foram polimórficos. Os fragmentos amplificados foram incluídos em uma matriz de dados para análise pelo método de Neighbor-Joining. No fenograma obtido, V. radiata e V. unguiculata ssp. cylindrica assumiram uma posição basal isolada dos subgrupos formados, enquanto que os acessos de V. unguiculata subdividiram-se em dois grupos, nos quais os acessos contrastantes quanto à tolerância ao caruncho se distribuíram em subgrupos distintos, o que evidencia a existência de diferenças genéticas significativas entre alguns candidatos. Nesse sentido, os resultados obtidos demonstraram que os ensaios de tolerância ao caruncho associados à aplicação de marcadores moleculares foram capazes de identificar genótipos contrastantes de feijão-caupi promissores para aplicação em trabalhos de melhoramento vegetal, bem como para a geração de populações segregantes adequadas ao mapeamento genético.

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Aceria guerreronis keifer is one of the main coconut pests in Brazil, its attack causes a premature fall, reduction of fruit size, water weight and volume, as well as stetic esthetic damages (necrosis) depreciating the product. Currently, there are five pesticides registered for the control of the coconut-necrotic mite. However, there is no specificity to A. guerreronis. The Lamiaceae family stands out with some specimens with notable insecticidal activity, among them the Vitex genus, known to have ecdysteroids and iridoids as markers of the genus. However, there is a shortage of specific chemotaxonomic studies for volatile compounds. The objective of this work was to identify the volatile compounds present in the essential oils of the species of Vitex gardneriana Schauer, Vitex capitata Vahl., Vitex rufescens A. Juss., Vitex megapotamica (Spreng.) Moldenk and to evaluate possible effects of toxicity and repellence against A. guerreronis. In addition to analyzing the genetic diversity of species of the genus Vitex using ISSR markers, verifying the occurrence of statistical correlations between the genetic diversity and the chemical profiles of the essential oils. Chromatographic and spectroscopic techniques (GC-MS / FID) were used for the detection of volatile compounds, and the molecular markers of ISSR for the genetic characterization of the species, as well as toxicity and repellency tests against necrotic necrosis. The results showed 46, 45, 41 and 37 constituents for V. capitata, V. megapotamica, V. gardneriana and V. rufences respectively, among which, -copaene, (E)-caryophyllene, -elemene, -humulene, trans-cadine-1(6)4-diene, viridiflorene and -cadinene were the main components of V. capitata and V. megapotamica. 6,9-guaiadiene, caryophyllene oxide, L-calamenene for V. gardneriana and (E)-cariophylene, ledol, germacrene D, -humulene, allo-aromadendrene, viridiflorene and -elemene for V. rufences. The results of molecular markers of ISSR generated a total of 90 fragments, of which 100% were polymorphic. The genotypes of the four Vitex species selected for this study were pooled using the Jaccard coefficient (JC). The mean similarity was 0.21 JC (0.08-0.45 JC). A clear separation of two groups was observed, with V. gardneriana being the most isolated of the four species, and V. capitata and V. megapotamica more closely related to each other. The acaricide activity of the four oils was also evaluated by means of toxicity and repellency tests against A. guerreronis. The essential oil of V. gardneriana showed acaricidal activity against A. guerreronis with a LC50 of 0.85 mg.mL-1, whereas the oils of V. capitata, V. megapotamica and V. rufecens were not toxic to this pest up to 2.3 mg. Ml-1 when tested through the toxicity bioassay. In general, essential oils distilled from the Vitex species repelled A. guerreronis after 24 hours at a concentration of 1.8 mg.mL-1 using abamectin as a control in the two-choice disc repellency assay, however, the essential oil of V. gardneriana presented a statistically significant acaricidal activity against the coconut mite necrosis. Introducing himself as a potential candidate for the development of an acaricide product. / O ácaro-da-necrose-de-coqueiro Aceria guerreronis keifer é uma das principais pragas de coqueiro do Brasil, seu ataque causa queda prematura, redução no tamanho e peso dos frutos e volume de água, além dos danos estéticos (necroses) na casca que diminui o valor comercial do produto. Atualmente, existem cinco agrotóxicos registrados para o controle dessa praga. No entanto, não há especificidade ao A. guerreronis. A família Lamiaceae se destaca com alguns exemplares com notória atividade inseticida, dentre eles, o gênero Vitex, conhecido por possuir dentre seus constituintes os ecdisteroides e iridoides como marcadores do gênero, todavia, há uma escassez de estudo quimiotaxonômicos específicos para compostos voláteis. O objetivo deste trabalho foi identificar os compostos voláteis presentes nos óleos essenciais das espécies de Vitex gardneriana Schauer, Vitex capitata Vahl., Vitex rufescens A. Juss., Vitex megapotamica (Spreng.) Moldenk e avaliar possíveis efeitos de toxicidade e repelência frente ao A. guerreronis, além de analisar a diversidade genética de espécies do gênero Vitex. Técnicas cromatográficas e espectroscópicas (CG-MS/FID) foram utilizadas para detecção dos compostos voláteis, e os marcadores moleculares de ISSR foram empregados para a caracterização genética das espécies, além de testes de toxicidade e repelência contra ao àcaro-da-necrose. Os resultados mostraram 46, 45, 41 e 37 constituintes para V. capitata, V. megapotamica, V. gardneriana e V. rufences respectivamente, dentre os quais, -copaeno, (E)-cariofileno, -elemeno, -humuleno, trans-cadina-1(6),4-dieno, viridiflorene e -cadineno foram os principais componentes de V. capitata e V. megapotamica. 6,9- guaiadieno, óxido de cariofileno, L-calameneno para V. gardneriana e (E)-cariofileno, ledol, germacreno D, -humuleno, allo-aromadendreno, viridifloreno e -elemene para V. rufences. Os resultados dos marcadores moleculares de ISSR geraram um total de 90 fragmentos, dos quais 100% foram polimórficos. Os genótipos das quatro espécies de Vitex selecionadas para este estudo foram agrupados utilizando o coeficiente Jaccard (JC). A média de similaridade foi de 0,21 JC (0,08-0,45 JC). Observou-se uma clara separação de dois grupos, sendo V. gardneriana a mais isolada das quatro espécies, e V. capitata e V. megapotamica mais próxima geneticamente entre si. A atividade acaricida dos quatro óleos também foi avaliada por meio de ensaios de toxicidade e repelência contra o àcaro-da-necrose. O óleo essencial de V. gardneriana mostrou atividade acaricida ao A. guerreronis com uma CL50 de 0,85 mg.mL-1, enquanto que os óleos de V. capitata, V. megapotamica e V. rufecens não foram tóxicos para esta praga até 2,3 mg.ml-1, quando testados através do bioensaio de toxicidade. Em geral, os óleos essenciais destilados a partir das espécies de Vitex repeliram o A. guerreronis após 24 horas a uma concentração de 1.8 mg.mL-1 utilizando abamectina como controle no ensaio de repelência disco de duas escolha, no entanto, o óleo essencial de V. gardneriana apresentou atividade acaricida estatisticamente significativa frente ao ácaro-da-necrose-do-coqueiro. Apresentando-se como potencial candidato para o desenvolvimento de um produto acaricida. / São Cristóvão, SE