Determinação da pureza varietal em lotes de sementes de milho através de marcadores morfológicos e microssatélites. / Determination of varietal purity in maize seed lots using morphological and microsatellites markers.

AUTOR(ES)

Nilza Patricia Ramos

DATA DE PUBLICAÇÃO

2004

RESUMO

Maize inbred lines seed production has been conducted to avoid the presence of varietal contamination since it compromises the subsequent multiplication phases within the commercial seed program. Contaminant seeds are detected through varietal and/or genetic purity determination tests which are usually based on morphological and biochemical markers, depending on the desired effectiveness. Thus, more efficient alternative approaches have been tested, with special emphasis on those based on DNA polymorphism. In that context, the main objective of this research was to compare the efficiency of morphological and microsatellite markers to evaluate varietal purity of maize inbred lines and to determine the sensitiveness of the microsatellite technique to detect the occurrence of contaminant genotypes in seed lots. There were used four inbred lines (L1, L2, L3 and L4) supplied by Dow AgroSciences Ltd., mixed to attain contamination levels of 0, 1, 2, 5, 10 and 100%. L1 and L3 were considered pure strains while L2 and L4 were treated as contaminant genotypes. For the morphological marker evaluation seed, seedling and plant descriptors at different development stages were used. Specific maize primers were used in the microsatellite technique and the DNA was isolated from samples of 100 seeds or 100 pairs of seedling leaves. The DNA amplification reactions were conducted through polymerase chain reaction, with amplification program especially designed for microsatellites, and for fragments resolution, 3.5% agarose and 6% polyacrilamyde gels were used. Successive mixtures among DNA of contaminant line and pure line (0%, 0.01%, 0.013%, 0.02%, 0.04%, 0.1%, 0.2%, 1%, 2%, 5%, 10%, and 100%) were performed to simulate contamination levels with the objective of verify the microssatellite sensitiveness in detecting the occurrence of contaminants. Morphological characteristics of the seeds, seedlings or plants were less reliable to detect contamination in maize inbred lines than the microsatellite technique; this provides more efficient and accurate evaluation of varietal purity of seed lots. Levels with 1% of contamination were detected by the use of seed and seedling leaf samples. The experiment with mixtures of DNA (L1 - L2 and L3 - L4) using microsatellite technique allowed the consistent detection of 0.1% contaminant DNA concentration. Thus, the determination of varietal purity in maize seed lots is more efficient by using microsatellite markers than morphological markers.

ASSUNTO(S)

marcador molecular linhagem vegetal molecular mark maize semente- qualidade seed- quality vegetal lineage milho

Documentos Relacionados

• Avaliação da pureza genética em sementes de milho utilizando marcadores microssatélites.
• Determinação da pureza varietal de sementes de soja com o auxílio de marcadores moleculares microssatélites
• Sensibilidade dos microssatélites para determinar a pureza varietal em sementes de milho
• Avaliação da variabilidade genética em Musa spp. utilizando marcadores microssatélites.
• Determinação da pureza varietal de sementes da cultivar de soja BRS Raimunda utilizando marcadores moleculares.