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Caracteriza??o agron?mica, molecular e fitoqu?mica de Eplingiella Harley & J.F.B. Pastore

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Previous issue date: 2015-06-29 / Conselho Nacional de Pesquisa e Desenvolvimento Cient?fico e Tecnol?gico - CNPq / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / Funda??o de Amparo ? Pesquisa do Estado da Bahia - FAPEB / Eplingiella fruticosa (Salzm. Ex Benth.) Harley & JFB Pastore is an aromatic species, native, occurring in six states in northeastern Brazil (Bahia, Sergipe, Pernambuco, Paraiba, Rio Grande do Norte and Cear?). Popularly known as "alecrim de vaqueiro", is commonly found in street markets of the region and used to combat pain and seizures. Reviews in mice and in vitro studies show analgesic activity, vasodilating, cardioprotetiva, anti-inflammatory and larvicidal of its essential oil and of different types of the leaves extract. Recent studies show great variability in essential oil chemical composition of E. fruticosa, related to soil and climatic conditions and different plant organs. Thus, the species has great potential for exploration both agronomic, and by pharmaceutical companies. The aim of this study was to evaluate the vegetative propagation capacity and characterize previously Eplingiella genotypes, through morphological, agronomic, phytochemicals and molecular data. In Chapter I, two experiments were conducted: the first tested the effect of three substrates and the second evaluated five concentrations of IBA and three periods of cultivation. The design was a randomized block design with four replications. We evaluated survival percentage (% S), percentage of rooted cuttings (% EE), root length (CRE), number of shoots (NBE), dry mass of leaves (MSF), root dry weight (MSR) and total dry matter (MST). In Chapter II, twelve genotypes were collected, propagated vegetatively and transplanted. Twelve months after transplantation were assessed 12 quantitative traits, eight morphological and agronomic four. In Chapter III, the total DNA was extracted, then 20 primers were tested, of which nine were selected because they have better electrophoretic profiles agarose gel (2%). The binary matrix was computed in GEOCOMPAR II. It is estimated the diversity of the genetic structure parameters and the data were subjected to Bayesian analysis, and Neighbor-joining dendrogram and principal component analysis (PCA) based on matrix of Nei distances. And in Chapter IV, samples of 100g of leaves each repetition per genotype were used in the essential oil hydrodistillation in Clevenger type apparatus for three hours, quantifying the content. The identification of the compounds and their contents was performed by GC (FID) and GC / MS data 15 and the major compounds were used in diversity analysis. They have been made to cluster analysis and canonical variables, using as dissimilarity measure the Mahalanobis distance (D2). In the first experiment of Chapter I, significant differences were found for CRE, NBE, MSF, MSR and MST, with the best performance for the commercial substrate. In the second, positive effects have been identified both the addition of AIB as the cultivation time on the CRE variables, NBE, MSF and MSR, reaching maximum increment to the estimated concentration of 1.5 g L-1, at 60 days of cultivation. In Chapter II, there was significant variation by F test (p <0.01) for the CF features, LF, CBD, CBE, LP, and MFF MSF. The genotypes formed two groups for almost all variables, by Scott-Knott test (p <005), except for LP, which formed three. The EF002 and EF003 genotypes presented the highest levels for almost all variables. There was the formation of three groups for both UPGMA and for the canonical variables (CV). The characteristics that most contributed to the formation of groups were CBE, MFF and CF. The genotypes EF002, EF003, EF005 and EF012 stood out because they have higher genetic distances. In CHAPTER III, primers produced 131 polymorphic bands. The diversity index of Nei (Ne) ranged between 0.31 and 0.39, while Shannon (I) ranged between 0.33 and 0.48. The percentage coefficient of genetic differentiation (Gst) was 0.29. In AMOVA most of the variation was within populations (69%), while among populations was 27% and 4% among species, indicating a good genetic structure. The average value of Fst was 0.175, demonstrating intermediate differentiation between populations. The structure of the Bayesian analysis method revealed three possibilities for the formation of groups (K = 2; = 6; 8 =;), however, it presented many migrants and high level of mixing individuals. The dendrogram generated by the Neighbor-Joining method confirmed the formation of two groups, with good support for major clades (100%). PCA analysis in the first two axis explained 21.06% of the total variation among populations. Finally, in Chapter IV, the genotypes were classified into four clusters: 1 - EF001 genotypes, EF006, EF007, EF008, EF010, EF011 and EF012 with E-caryophyllene and bicyclogermacrene as major; 2 - EF002 and EF003 genotypes, with the majority same as the previous group, however, percentage with average about 30% higher; 3 - EF004 and EF005 genotypes that showed a greater production of E-caryophyllene; and 4 - with EF009 genotype, forming a single group to present ?-pinene as balanced majority and percentage among the rest. This result was confirmed by canonical variables, which explained 76% of the variation. The bicyclogermacrene compounds, 1,8-cineol, ?-copaene and spathulenol represented the most important variables for analysis. / Eplingiella fruticosa (Salzm. ex Benth.) Harley & J.F.B. Pastore ? uma esp?cie arom?tica, nativa, que ocorre em seis estados do nordeste brasileiro (Bahia, Sergipe, Pernambuco, Para?ba, Rio Grande do Norte e Cear?). Popularmente conhecida como ?alecrim de vaqueiro?, ? comumente encontrada em feiras livres da regi?o e utilizada no combate a dores e convuls?es. Avalia??es em camundongos e in vitro comprovam atividades analg?sicas, vasodilatadora, cardioprotetiva, antinflamat?ria e larvicida do seu ?leo essencial e de diferentes tipos de extrato de suas folhas. Estudos recentes apontam grande variabilidade na composi??o qu?mica do ?leo essencial de E. fruticosa, relacionada ?s condi??es edafoclim?ticas e aos diferentes ?rg?os vegetais. Sendo assim, a esp?cie apresenta grande pot?ncial de explora??o tanto agron?mica, quanto por ind?strias farmac?uticas. O objetivo geral deste estudo foi avaliar a capacidade de propaga??o vegetativa e caracterizar, previamente, gen?tipos de E. fruticosa, por meio de dados morfol?gicos, agron?micos, fitoqu?micos e moleculares. No CAP?TULO I, foram conduzidos dois experimentos: o primeiro testou o efeito de tr?s substratos e o segundo avaliou cinco concentra??es de AIB e tr?s per?odos de cultivo. O delineamento foi em blocos casualizado, com quatro repeti??es. Avaliou-se percentagem de sobreviv?ncia (%S), percentagem de estacas enraizadas (%EE), comprimento da raiz (CRE), n?mero de brota??es (NBE), massa seca de folhas (MSF), massa seca de raiz (MSR) e massa seca total (MST). No CAP?TULO II, doze gen?tipos foram coletados, propagados vegetativamente e transplantados. Doze meses ap?s o transplante foram avaliadas 12 caracter?sticas quantitativas, sendo oito morfol?gicase quatro agron?micas. No CAP?TULO III, o DNA total foi extra?do, em seguida 20 iniciadores foram testados, dos quais nove foram selecionados por apresentarem melhores perfis eletrofor?ticos em gel de agarose (2%). A matriz bin?ria foi computada no GEOCOMPAR II. Estimou-se os par?metros de diversidadee a estrutura gen?tica os dados foram submetidos ? an?lise Bayesiana, al?m de dendrograma Neighbor-joining e an?lise de componentes principais (PCA) com base na matriz de dist?ncias de Nei. E no CAP?TULO IV, amostras de 100g de folhas de cada repeti??o por gen?tipo foram utilizadas na hidrodestila??o do ?leo essencial, em aparelho tipo clevenger, durante tr?s horas, quantificando-se o teor. A identifica??o dos compostos e seus teores foi realizada por CG (DIC) e CG/EM e os dados de 15 compostos majorit?rios foram utilizados nas an?lises de diversidade. Foram procedidas an?lise de agrupamento e de vari?veis can?nicas, utilizando como medida de dissimilaridade a dist?ncia generalizada de Mahalanobis (D2).No primeiro experimento do CAP?TULO I, foram verificadas diferen?as significativas para CRE, NBE, MSF, MSR e MST, com melhor desempenho para o substrato comercial. No segundo, foram identificados efeitos positivos tanto da adi??o de AIB quanto dos tempos de cultivo sobre as vari?veis CRE, NBE, MSF e MSR, atingindo incremento m?ximo com a concentra??o estimada de 1,5 g L-1, aos 60 dias de cultivo. No CAP?TULO II, houve varia??o significativa, pelo teste de F (p<0,01), para as caracter?sticas CF, LF, CBD, CBE, LP, MFF e MSF. Os gen?tipos formaram dois grupos para quase todas as vari?veis, pelo teste de Scott-Knott (p<005), exceto para LP, que formou tr?s. Os gen?tipos EF002 e EF003 apresentaram as maiores m?dias para quase todas vari?veis. Houve a forma??o de tr?s grupos, tanto para UPGMA quanto para as vari?veis can?nicas (VC). As caracter?sticas que mais contribu?ram para a forma??o dos grupos foram CBE, MFF e CF. Os gen?tipos EF002, EF003, EF005 e EF012 se destacaram por apresentarem maiores dist?ncias gen?ticas. No CAP?TULO III, os iniciadores produziram 131 bandas polim?rficas. O ?ndice de diversidade de Nei (Ne) variou entre 0,31 e 0,39, enquanto Shannon (I) variou entre 0,33 e 0,48. O percentual do coeficiente de diferencia??o gen?tica (Gst) foi de 0,29. Na AMOVA a maior parte da varia??o ficou dentro das popula??es (69%), enquanto entre popula??es foi de 27% e entre esp?cies de 4%, indicando uma boa estrutura??o gen?tica. O valor m?dio de Fst foi 0,175, demonstrando diferencia??o intermedi?ria entre as popula??es. As an?lises de estrutura pelo m?todo Bayesiano revelou tr?s possibilidades de forma??o de grupos (K=2;=6;=8;), no entanto, apresentou muitos indiv?duos migrantes e elevado n?vel de miscigena??o. O dendograma gerado pelo m?todo de Neighbor-Joining confirmou a forma??o de dois grupos, com boa sustenta??o para os principais clados (100%). Na an?lise de PCA os dois primeiros axis explicaram 21,06% da varia??o total entre as popula??es. Por fim, no CAP?TULO IV, os gen?tipos foram classificados em quatro clusters: 1 - gen?tipos EF001, EF006, EF007, EF008, EF010, EF011 e EF012, com E-cariofileno e biciclogermacreno como majorit?rios; 2 - gen?tipos EF002 e EF003, com os mesmos majorit?rios que o grupo anterior, no entanto, com percentuais m?dios cerca de 30% superiores; 3 - gen?tipos EF004 e EF005, que evidenciaram uma maior produ??o de E-cariofileno; e 4 - com gen?tipo EF009, formando um grupo isolado por apresentar ?-pineno como majorit?rio e percentuais equilibrados entre os demais. Esse resultado foi confirmado pelas Vari?veis Can?nicas, que explicou 76% da varia??o. Os compostos biciclogermacreno, 1,8-cineol, ?-copaeno e espatulenol representaram as vari?veis de maior import?ncia para a an?lise.

Identiferoai:union.ndltd.org:IBICT/oai:tede2.uefs.br:8080:tede/293
Date29 June 2015
CreatorsSilva, Anderson de Carvalho
ContributorsOliveira, Lenaldo Muniz de
PublisherUniversidade Estadual de Feira de Santana, Doutorado Acad?mico em Recursos Gen?ticos Vegetais, UEFS, Brasil, DEPARTAMENTO DE CI?NCIAS BIOL?GICAS
Source SetsIBICT Brazilian ETDs
LanguagePortuguese
Detected LanguageEnglish
Typeinfo:eu-repo/semantics/publishedVersion, info:eu-repo/semantics/doctoralThesis
Formatapplication/pdf
Sourcereponame:Biblioteca Digital de Teses e DissertaƧƵes da UEFS, instname:Universidade Estadual de Feira de Santana, instacron:UEFS
Rightsinfo:eu-repo/semantics/openAccess
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