Articles
USE OF ISOLATED PROTOPLASTS IN ROSE BREEDING
Article number
547_4
Pages
35 – 44
Language
Abstract
A combination of 1.5% Cellulysin, 0.5% Driselase and 0.5% Macerase allows the isolation of protoplasts out of in vitro shoot cultures, callus and, preferentially, out of non embryogenic as well as embryogenic cell suspension cultures from different genotypes.
Three factors were found to distinctly influence protoplast yields from cell cultures, i.e. (a) the type of auxin in the preculture media, (b) light conditions during incubation of the source material, and (c) the appropriate phase within the growth cycle of suspensions.
Protoplasts isolated from non embryogenic cell suspension cultures of different rose species regenerate non morphogenic callus with the exception of R. persica x R. xanthina, which generated plants.
In contrast, protoplasts isolated from embryogenic cell suspension cultures regularly give rise to an embryogenic type of callus, from which plantlets were regenerated in cultivars ‘Heckenzauber’ and ‘Pariser Charme’. Experiments with the GFP-gene as reporter were performed in order to establish a protocol for direct transformation of protoplasts by PEG mediation.
It became evident, that the genotype as well as the incubation conditions during preculture of the cell suspensions used for protoplast isolation both influence the transformation rates.
Stable transformation was obtained, but so far incorporation of the GFP-gene inhibited cell divisions.
Non embryogenic cell suspension cultures were established from different wild rose species with resistance against blackspot (Diplocarpon rosae) to be used for somatic hybridization with susceptible rose cultivars.
Asymmetric and symmetric fusion of protoplasts is performed by PEG mediation after treatment of protoplasts with iodacetate, rhodamine-6-G or X-rays.
Hybrid callus has been regenerated and transfered to somatic embryo induction media.
Three factors were found to distinctly influence protoplast yields from cell cultures, i.e. (a) the type of auxin in the preculture media, (b) light conditions during incubation of the source material, and (c) the appropriate phase within the growth cycle of suspensions.
Protoplasts isolated from non embryogenic cell suspension cultures of different rose species regenerate non morphogenic callus with the exception of R. persica x R. xanthina, which generated plants.
In contrast, protoplasts isolated from embryogenic cell suspension cultures regularly give rise to an embryogenic type of callus, from which plantlets were regenerated in cultivars ‘Heckenzauber’ and ‘Pariser Charme’. Experiments with the GFP-gene as reporter were performed in order to establish a protocol for direct transformation of protoplasts by PEG mediation.
It became evident, that the genotype as well as the incubation conditions during preculture of the cell suspensions used for protoplast isolation both influence the transformation rates.
Stable transformation was obtained, but so far incorporation of the GFP-gene inhibited cell divisions.
Non embryogenic cell suspension cultures were established from different wild rose species with resistance against blackspot (Diplocarpon rosae) to be used for somatic hybridization with susceptible rose cultivars.
Asymmetric and symmetric fusion of protoplasts is performed by PEG mediation after treatment of protoplasts with iodacetate, rhodamine-6-G or X-rays.
Hybrid callus has been regenerated and transfered to somatic embryo induction media.
Authors
A. Schum, K. Hofmann
Keywords
somatic hybridization, direct transformation, blackspot resistance, Rosa wichuraiana, Rosa multiflora, Rosa roxburghii
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