Molecular evidence of true-to-type propagation of a 3-year-old Norway spruce through somatic embryogenesis.

Some progress has recently been made in establishing a system enabling somatic embryos to be initiated from old elite trees. We report here the first results demonstrating the molecular conformity of somatic embryos initiated from increasingly old Norway spruce (Picea abies (L.) Karst.), as indicated by an analysis of six nuclear microsatellites that showed an extremely high tendency to mutate during in vitro culture. No allelic difference was detected at these loci among plants regenerated from somatic embryos or between the former and mother plants. Moreover, phenotypical data acquired on the same 3- to 9-year-old plants growing in the field sampled for molecular analyses were totally in accord with the results on molecular conformity.

Two highly divergent 5S rDNA unit size classes occur in composite tandem array in European larch (Larix decidua Mill.) and Japanese larch (Larix kaempferi (Lamb.) Carr.).

The 5S ribosomal DNA unit structure and organization have been investigated in Larix decidua and Larix kaempferi using selective amplification of gene and spacer, sequence analysis and homologous probe hybridization. Two highly divergent unit size classes of approximately 650 and 870 bp were detected in both species. Sequence analysis in Larix decidua revealed that length variations occur in the middle spacer region and are the result of duplications (in the long spacers) and considerable sequence heterogeneity. Conversely, the transcribed region is of uniform length (120 bp), and the nucleotide sequence of one Larix decidua clone is similar to that reported for other gymnosperms. Sequence comparison of the larch spacers with two other Pinaceae species (Pinus radiata and Picea glauca) showed that the 5' and 3' regions flanking the gene (40 and 60 bp, respectively) are quite conserved, suggesting a regulatory role. Moreover, a small element of about 70 bp located in the middle spacer region was found to be common to the larch long units and the six Pinus radiata spacer clones previously sequenced (64% sequence identity). The short and long unit size classes are mainly organized in composite tandem array(s) with evidence of extensive zones of strict alternation in both species. Mechanisms underlying this unusual association of divergent units in larch 5S rDNA arrays are discussed.