Long-term subculture randomly affects morphology and subsequent maturation of early somatic embryos in maritime pine

Autor: Jean-François Trontin, David Breton, Luc Harvengt, Alain Bouvet, Jean-Michel Favre
Rok vydání: 2006
Předmět:
Zdroj: Plant Cell, Tissue and Organ Culture. 87:95-108
ISSN: 1573-5044
0167-6857
DOI: 10.1007/s11240-006-9144-9
Popis: Somatic embryogenesis is affected by highly variable maturation yields in Pinus pinaster. Origins of this variability were investigated by testing effects of spatial and temporal division of initial embryogenic tissue into independently cultivated pieces. One embryogenic cell-line was proliferated to obtain six embryonal-suspensor masses (ESM) treated as six sub-lines within a single dish. After proliferation to reach 2, 4 and 8 dishes (12, 24, 48 ESM), the 48 ESM (six sub-lines) were maintained by weekly subculture (34 weeks). ESM observations and maturation tests were regularly performed during the amplification and maintenance periods. Relations between maturation yields as well as cotylfedonary embryo length and immature embryo morphology were analyzed. ESM yielded variable results, independently from culture spatial (dish) and temporal (sub-line) subdivision. Maturation yields and length of regenerated embryos globally decreased as a function of subculture number. Concomitantly, morphological degradation of immature embryos occurred, indicating a global loss of embryogenic ability. Maturation variability probably results from immature embryos heterogeneity, which could be increased by manipulations during subculture. This is the first time that this evolution along time in culture is precisely described in conifers somatic embryos.
Databáze: OpenAIRE
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