Microsatellite analysis of embryogenic lines of the Norway spruce Picea abies (L.) Karst.

The method of somatic embryogenesis could be used for industrial production of clonal planting material. For this reason special attention is paid to the studies of various genetic disorders that arise in vitro because of their possibility to be inherited by the vegetative offsprings. In our investigation the levels of somaclonal variability and mixoploidy in callus colonies and plantlets of six embryogenic lines of Norway spruce (Picea abies (L.) Karst.) were studied. ½ LM nutrient medium supplemented with sucrose and glutamine was used at different stages of plant tissues and plantlets in vitro cultivation. Growth regulators 2,4-D, 6-BAP, ABA, IMC and activated carbon were added to nutrient media if necessary. Microsatellite analysis of samples was carried out using five EST-SSR markers Pa28, Pa33, Pa56, Pa47, Pa52. It was shown that in samples of three of the six embryogenic lines, more than two (three or four) allelic variants were identified at the Pa28 locus. In a number of cases, the tissues studied have pronounced signs of mixoploidy. Appropriate microsatellite analysis of samples collected from 33 middle-aged trees of Norway spruce from naturally formed forest stand was additionally carried out in order to compare the results with the data obtained earlier. Only two plants showed the presence of triploid cells, indicating a significantly lower level of genomic or chromosomal aberrations than observed in in vitro cultures.

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