Effective micropropagation of mature aspen: Use in breeding.

Malá J., Máchová P., Cvrčková H., Čížková L., Soudek P.
ACTA HORTICULTURAE 764: 239-246, 2007

Klíčová slova: aspen, micropropagation, Agrobacterium tumefaciens
Abstrakt: Aspen (Populus tremula) and its hybrid, (Populus tremula × P. tremuloides) represent very modest tree species in climatic conditions and soil quality. Aspen clones and their synthetic populations are very suitable for planting on agricultural soils (lignicultures) and as the pioneering woody species for immission clearings. The first aim of the research was to verify a method of rapid in vitro reproduction of aspen. Mature aspen trees growing in experimental outside trial plots proved to be convenient for growth in extreme sites and for production of quality timber. Moreover, the standardized rapid micropropagation could be effectively used for transformation procedures by Agrobacterium tumefaciens in aspen. The primary cultures were established from 24 years old trees from the provenance plots. Microcuttings from multi-apex cultures were used for rooting. The losses during rooting and acclimatization were minimal, around 2%. The micropropagated aspen plantlets are just growing on the trial plots. Twenty-five selected aspen clones are established in gene bank. The second aim of the study was to verify the transformation ability of micropropagated selected clone of hybrid aspen. Four gene constructs of A. tumefaciens (contained NPTII and GUS gene) comprising different promotors were used for transformations of aspen hybrid stem segments. A promotor for successful transformations was selected on the basis of GUS activity assay. The highest activity was estimated in transformants with construct pBI 121, comprising the reporter gene GUS controlled by promotor CAMV 35S.
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