hrvatski

Procjena homogenosti hibrida kukuruza mikrosatelitskim markerima

Microsatellites (SSRs, Short Sequence Repeats) consist of tandemly repeated units up to 10 base-pairs in length. They are widely dispersed in eukaryotic genomes, they show Mendelian inheritance and are often highly polymorphic. The objective of this study was to establish a quick and reliable procedure in order to distinguish between heterozygous hybrid maize and its homozygous parents. The seeds of parental lines OS 438-95 (mother), OS 1-44 (father) and the hybrid were grown in the wet chamber for six days. The seedlings were harvested and DNA isolated according to Doyle & Doyle (1996). Initial screening of parental lines for polymorphic SSR loci was performed using 12 primer pairs. Two of the polymorphic loci were chosen, DNA from single hybrid plants amplified and the PCR products separated using agarose gel electrophoresis. DNA fragments were visualised using ethidium-bromide staining. Four out of 12 primers used for initial screening showed polymorphisms in parental lines. Primers phi 128 and phi 037 which produced clearly visible bands on the agarose gels were chosen for the subsequent PCR amplification of the corresponding SSR loci from the hybrid plants. Out of 88 hybrid plants, 86 showed the same di-allelic pattern with phi 037, while two amplifications were unsuccessful. For phi 128, a complete homogeneity was observed. For the two SSR loci examined, the hybrid showed almost 100% homogeneity. The procedure applied here represents a quick and simple approach for the hybrid maize genetic purity evaluation, detecting homozygous seeds which are the most common impurity in commercial hybrids.

kukruz; inbred linije; mikrosatelitski markeri; procjena čistoće

nije evidentirano

engleski

Maize hybrid homogeneity evaluation using microsatellite markers

Microsatellites (SSRs, Short Sequence Repeats) consist of tandemly repeated units up to 10 base-pairs in length. They are widely dispersed in eukaryotic genomes, they show Mendelian inheritance and are often highly polymorphic. The objective of this study was to establish a quick and reliable procedure in order to distinguish between heterozygous hybrid maize and its homozygous parents. The seeds of parental lines OS 438-95 (mother), OS 1-44 (father) and the hybrid were grown in the wet chamber for six days. The seedlings were harvested and DNA isolated according to Doyle & Doyle (1996). Initial screening of parental lines for polymorphic SSR loci was performed using 12 primer pairs. Two of the polymorphic loci were chosen, DNA from single hybrid plants amplified and the PCR products separated using agarose gel electrophoresis. DNA fragments were visualised using ethidium-bromide staining. Four out of 12 primers used for initial screening showed polymorphisms in parental lines. Primers phi 128 and phi 037 which produced clearly visible bands on the agarose gels were chosen for the subsequent PCR amplification of the corresponding SSR loci from the hybrid plants. Out of 88 hybrid plants, 86 showed the same di-allelic pattern with phi 037, while two amplifications were unsuccessful. For phi 128, a complete homogeneity was observed. For the two SSR loci examined, the hybrid showed almost 100% homogeneity. The procedure applied here represents a quick and simple approach for the hybrid maize genetic purity evaluation, detecting homozygous seeds which are the most common impurity in commercial hybrids.

maize; inbred lines; microsatellite markers; homogeneity evaluation

nije evidentirano