Somatic embryogenesis and somaclonal variation in Norway spruce: Morphogenetic, cytogenetic and molecular approaches

Four embryogenic clones of Norway spruce have been subcultivated and observed over several years to determine the evolution of production of mature embryos and to assess the quality of the embryos produced. A wide range of intraclonal quantitative and qualitative variability has been observed within this production. Certain morphologic deviations appeared at the immature stage and after maturation, such as immature embryos with a diffuse organization, complete or part albino mature embryos or acclimated somatic seedlings comparable to dwarf mutants. All of these phenotypic variations could be the result of a modification of the genome itself or of only the expression of the genome. Two approaches, chromosome counting and RAPD (random amplified polymorphic DNA), were chosen for their capacity to detect genotypic variations: respectively, genomic and chromosomic or genic mutations. The cytogenetic approach revealed, for the first time in this species, three cases of mutated acclimated somatic plants: one totally trisomic and two chimeras with trisomic buds and diploid roots. Other cases of 5-year-old trisomic, double trisomic, tetraploid or mixoploid embryogenic masses were also detected. The molecular approach (RAPD) revealed no somaclonal variation despite the large sample of DNA and primers used and the important interclonal variation observed