ABSTRACT
Morphological and agronomical describers are traditionally used in plant characterization. However, the usage of these describers have some limitations such as susceptibility to abiotic and biotic stress and environmental factors. Furthermore, the describers are not stable over time and many can only be evaluated during the adult phase of the plants, which requires time and physical space. Molecular markers offer numerous advantages compared to the conventional alternatives based on phenotype: they are stable and detectable in all vegetable tissues, and are independent of the environment and development phase. One of the main advantages of the use of molecular markers is the time reduction in the identification of genetic diversity among the studied subjects, as the genotypes may even be described for the seed or seedling phase. Many countries have already adopted molecular markers to identify olive cultivars more accurately. The aim of this study was to evaluate the genetic identity of eight olive accessions supposedly belonging to cultivar Arbequina by using microsatellite (SSR) and Sequence Characterized Amplified Region (SCAR) markers. One accession corresponding to the cultivar was also incorporated into the analysis as a reference genotype. The molecular marker data were analyzed on the software GENALEX6.The markers generated an accumulated PI and PE of 1.26 x 10-6 and 0.949, respectively.The results supported the hypothesis that all accessions belong to the cultivar Arbequina, and the markers can therefore be applied to other varieties of olive species.
Descritores morfológicos e agronômicos são tradicionalmente utilizados na caracterização de plantas. Apesar de recomendado, o emprego destes descritores apresenta algumas limitações como a influência a estresses abióticos e bióticos e aos efeitos do ambiente. Além disso, não são estáveis ao longo do tempo e muitos só podem ser avaliados durante a fase adulta das plantas, o que requer tempo e espaço físico para as avaliações. Os marcadores moleculares oferecem numerosas vantagens relativamente às alternativas convencionais baseadas no fenótipo, pois são estáveis e detectáveis em todos os tecidos vegetais, independente do ambiente e fase de desenvolvimento e uma das principais vantagens da utilização destes é proporcionar a redução do tempo na identificação da diversidade genética entre os indivíduos trabalhados, podendo ser avaliadas genótipos ainda na fase de semente ou de plântula. O objetivo deste estudo foi avaliar a identidade genética de oito acessos de oliveira supostamente pertencentes a cultivar Arbequina usando microssatélites (SSR) e Sequence Characterized Amplified Region (SCAR) marcadores. Um acesso correspondente a cultivar também foi incorporado na análise como o genótipo de referência. Os dados de marcadores moleculares foram analisados com o software GENALEX 6. Como resultado, os marcadores SSR geraram um PI acumulada e PE de 1,26 x 10- 6 e 0,949, respectivamente. Os resultados suportam a hipótese de que todos os acessos pertencem a cultivar Arbequina, e, por conseguinte, esses marcadores podem ser aplicados em situações semelhantes em outras variedades de espécies de oliveira.
Subject(s)
DNA Fingerprinting , OleaABSTRACT
To accelerate the identification of HLA-DPB1 matched marrow donors from unrelated population, a very simple HLA-DPB1 genotyping method called PCR-fingerprinting (PCRF) was developed according to the theory about homoduplex and heteroduplex formation from different PCR coding strands and non-coding ones. Unlike PCR-SSCP, strict laboratory condition is not needed in the PCRF. After denaturing at 94℃ for 2 min and cooling at 37℃ for 8 min, the PCR product was separated by 8% PAGE for 5 h and polymorphism band patterns would appear when the gel staining was completed with either EB or silver staining procedure. To confirm its reliability, 21 individuals from 9 family whose DPB1 genotypes assigned by PCR-RFLP were verified. It was found that there were 8 PCRF patterns corresponding to the 9 HLA-DPB1 genotypes from the 21 cases and the same DPB1 genotypes produced identical PCRF pattern except one pair.The factors on efficient separation of heteroduplexes and homoduplexes were also discussed.