Estudo da diversidade genética e química de Uncaria tomentosa Willd. ex Roem. & Schult. e Uncaria guianensis Gmell. de populações naturais localizadas na Amazônia
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2016-03-11
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Universidade Estadual Paulista (Unesp)
Resumo
Uncaria tomentosa Willd. ex Roem. & Schult. e Uncaria guianensis Gmell. são plantas medicinais nativas da Amazônia, utilizadas na medicina tradicional com ação anti-inflamatória, sofrem pressão antrópica como o desmatamento e extrativismo predatório para uso e necessitam de cuidados em relação à conservação. U. tomentosa é uma das plantas medicinais que compõe a lista da relação nacional de medicamentos essenciais (RENAME) disponibilizada pelo Ministério da Saúde para os municípios brasileiros, através do Sistema Único de Saúde (SUS) e a U. guianensis apesar de não estar nesta lista apresenta também alcaloides oxindólicos pentacíclicos, como marcadores químicos, sendo utilizada popularmente como anti-inflamatória e sua ocorrência na natureza é maior quando comparada à U. tomentosa. Os objetivos desse trabalho foram avaliar a diversidade genética e química entre os indivíduos de U. tomentosa e U. guianensis por marcador molecular SRAP (Sequence-Related Amplifield Polymosphism) e quantificar os alcaloides oxindólicos pentacíclicos mitrafilina e isomitrafilina em folhas por HPLC. A coleta do material foi feita nos estados do Acre, Amazonas, Amapá e Pará. Para ambas as espécies foram coletadas oito populações, em municípios distintos com 20 indivíduos cada. A genotipagem de U. tomentosa foi realizada utilizando três combinações de primers e os fragmentos submetidos à eletroforese em gel de poliacrilamida. A genotipagem de U. guianensis foi realizada usando quatro combinações de primers e os fragmentos analisados no equipamento 4300 DNA Analyser LI-COR®. A quantificação dos alcaloides oxindólicos pentacíclicos mitrafilina e isomitrafilina foi realizada de acordo com método já descrito na literatura com algumas modificações. Nas populações avaliadas de U. tomentosa a variabilidade genética foi maior dentro (75%) das populações do que entre elas (25%). O valor de Fst foi de 0,246, evidenciando que as populações estão estruturadas em Modelo de Ilhas. A maior porcentagem de locus polimórficos (95,68%) e a maior diversidade genética foram encontradas na população coletada no município de Mâncio Lima - AC. O fluxo gênico foi considerado restrito (Nm=1,57) e não houve correlação entre as distâncias geográfica e genética. Para a espécie U. guianensis, a variabilidade genética também foi maior dentro das populações (81%) do que entre elas (19%). O valor de Fst foi de 0,188, o que mostra que as populações estão se aproximando do Modelo de Ilhas. A maior porcentagem de locus polimórficos (90,21%) e a maior variabilidade genética foram verificadas na população coletada no município de Mazagão-AC. O fluxo gênico foi considerado baixo (Nm = 2,57) e houve fraca correlação entre as distâncias geográfica e genética, portanto considerada não significativa. A maior concentração dos alcaloides mitrafilina (11,17 mg.g-1.PS) e isomitrafilina (2,99 mg.g-1.PS) em U. tomentosa foram encontradas nos indivíduos da população de Tarauacá-AC e em U. guianensis foi de 1,09 mg.g-1.PS de mitrafilina e 0,29 mg.g-1.PS de isomitrafilina encontradas na população de Boca do Acre-AM. Além disso, para essa espécie, foi possível quantificar mitrafilina em apenas 24,8% dos indivíduos e a isomitrafilina em 20,4% dos indivíduos estudados. Conclui-se que a conservação de ambas as espécies deverá ser realizada com a coleta de um maior número de indivíduos nas populações com maiores variabilidades genética e química.
Uncaria tomentosa Willd. ex Roem. & Schult. and Uncaria guianensis Gmell. are medicinal plants natives from Amazonia state, used in the ocidental medicine for its anti-inflammatory effect, which are suffering from the actions of mankind by deforestation and predatory extraction, requesting care for their conservation. U. tomentosa is one of the medicinal plants that make up the national ratio of the list of essential drugs (RENAME) made available by the Ministry of Health to the municipalities through the Unified Health System (SUS) and U. guianensis although this is not list also features pentacyclic oxindole alkaloids, as markers, being popularly used as anti-inflammatory and its occurrence in nature is higher compared to U. tomentosa. The objectives of the present study were to evaluate the chemical and genetical diversity between individuals of U. tomentosa and U. guianensis by molecular marker SRAP (Sequence-Related Amplifield Polymosphism) and quantify pentacyclic oxindolic alkaloides mitraphylline and isomitraphylline in leaves via HPLC. Samples were collected in the states of Acre, Amazonas, Amapá and Pará. For both species, 20 samples were collected from each of the eight populations coming from different locations. The genotyping of U. tomentosa was realized by using the combination of three primers and the fractions submitted to an electrophoresis in acrylamide gel. The genotyping of U. guianensis was realized by using the combination of four primers and the fractions analyzed with a 4300 DNA Analyser LICOR®. The quantification of pentacyclic oxindole alkaloides mitraphylline and isomitraphylline were realized by using the existing method with some modifications. The U. tomentosa populations showed higher genetic variability inside the same population(75%) than between different populations (25%). The value of Fst was 0,246, showing that the populations are following the Island Model. The highest percentage of polymorphic locus (95,68%) and the highest genetic diversity were found in the population from Mâncio Lima - AC. The gene traffic was considered restrictive (Nm=1,57) and there was no correlation between geographic and genetic distances. For U. guianensis, the genetic variability was higher inside same populations (81%) than between different populations (19%). The value of Fst was 0,188, showing that the populations are coming close to the Island Model (modelo de ilhas). A higher percentage of polymorphic locus (90,21%) and a higher genetic variability were confirmed in the population colected in Mazagão - AC. The gene traffic was considered low (Nm = 2,57) and as having a weak correlation between geographic and genetic distances and, therefore, considered not significant. The highest alkaloid concentrations of mitraphylline (11,17 mg.g-1 .PS) and isomitraphylline (2,99 mg.g-1 .PS) in U. tomentosa were found in samples of the population from Tarauacá - AC and in U. guianensis samples showed alkaloid concentrations of 1,09 mg.g-1 .PS of Mitraphylline and 0,29 mg.g-1 .PS of Isomitraphylline in the population from Boca do Acre - AM. Besides that, to this species, it was possible to quantify mitraphylline in only 24.8% of individuals isomitraphylline and 20.4% of subjects studied. It’s concluded that the conservation of both species should be done with the collection of a greater number of individuals in populations with higher genetic and chemical variability.
Uncaria tomentosa Willd. ex Roem. & Schult. and Uncaria guianensis Gmell. are medicinal plants natives from Amazonia state, used in the ocidental medicine for its anti-inflammatory effect, which are suffering from the actions of mankind by deforestation and predatory extraction, requesting care for their conservation. U. tomentosa is one of the medicinal plants that make up the national ratio of the list of essential drugs (RENAME) made available by the Ministry of Health to the municipalities through the Unified Health System (SUS) and U. guianensis although this is not list also features pentacyclic oxindole alkaloids, as markers, being popularly used as anti-inflammatory and its occurrence in nature is higher compared to U. tomentosa. The objectives of the present study were to evaluate the chemical and genetical diversity between individuals of U. tomentosa and U. guianensis by molecular marker SRAP (Sequence-Related Amplifield Polymosphism) and quantify pentacyclic oxindolic alkaloides mitraphylline and isomitraphylline in leaves via HPLC. Samples were collected in the states of Acre, Amazonas, Amapá and Pará. For both species, 20 samples were collected from each of the eight populations coming from different locations. The genotyping of U. tomentosa was realized by using the combination of three primers and the fractions submitted to an electrophoresis in acrylamide gel. The genotyping of U. guianensis was realized by using the combination of four primers and the fractions analyzed with a 4300 DNA Analyser LICOR®. The quantification of pentacyclic oxindole alkaloides mitraphylline and isomitraphylline were realized by using the existing method with some modifications. The U. tomentosa populations showed higher genetic variability inside the same population(75%) than between different populations (25%). The value of Fst was 0,246, showing that the populations are following the Island Model. The highest percentage of polymorphic locus (95,68%) and the highest genetic diversity were found in the population from Mâncio Lima - AC. The gene traffic was considered restrictive (Nm=1,57) and there was no correlation between geographic and genetic distances. For U. guianensis, the genetic variability was higher inside same populations (81%) than between different populations (19%). The value of Fst was 0,188, showing that the populations are coming close to the Island Model (modelo de ilhas). A higher percentage of polymorphic locus (90,21%) and a higher genetic variability were confirmed in the population colected in Mazagão - AC. The gene traffic was considered low (Nm = 2,57) and as having a weak correlation between geographic and genetic distances and, therefore, considered not significant. The highest alkaloid concentrations of mitraphylline (11,17 mg.g-1 .PS) and isomitraphylline (2,99 mg.g-1 .PS) in U. tomentosa were found in samples of the population from Tarauacá - AC and in U. guianensis samples showed alkaloid concentrations of 1,09 mg.g-1 .PS of Mitraphylline and 0,29 mg.g-1 .PS of Isomitraphylline in the population from Boca do Acre - AM. Besides that, to this species, it was possible to quantify mitraphylline in only 24.8% of individuals isomitraphylline and 20.4% of subjects studied. It’s concluded that the conservation of both species should be done with the collection of a greater number of individuals in populations with higher genetic and chemical variability.
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Palavras-chave
Unha-de-gato, Conservação, SRAP, Plantas medicinais, Alcaloides, Cat’s clow, Conservation, Medicinal plants, Alkaloids