During and subsequent to digestion of the cell wall, the protoplast becomes very sensitive to osmotic stress. This means cell wall digestion and protoplast storage must be done in an isotonic solution to prevent rupture of the plasma membrane. Uses for protoplasts[ edit ] Fused protoplast on left , containing both chloroplasts from a leaf cell as well as a coloured vacuole from a petal. Protoplasts can be used to study membrane biology, including the uptake of macromolecules and viruses. These are also used in somaclonal variation. Protoplasts are widely used for DNA transformation for making genetically modified organisms , since the cell wall would otherwise block the passage of DNA into the cell.
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Molecular characterization of inter- and intra-specific somatic hybrids of potato using randomly amplified polymorphic DNA RAPD markers. Regeneration of fertile plants from Helianthus nuttalli and Helianthus giganteus L.
Many wild Solanum species are regarded as important sources protoplasos disease resistance and tolerance to many abiotic stresses Hawkes, but their use in potato breeding is limited due to poor crossability and sterility of interspecific hybrids. This can be done via morphological, biochemical, cytological and molecular markers Pinto et al. It has been demonstrated that protoplast-derived d show a higher degree of chromosomal number variation than those derived from tissue or organ culture.
Interspecific somatic hybridization between Solanum tuberosum L. Protoplasts density is usually determined by protoplastow. The mixture was incubated for thirty minutes and then protoplasts were collected by centrifugation at rpm during five minutes. Somatic hybridization in the genus Solanum: Tissue culture response in various wild and cultivated Solanum germplasm accessions for exploitation in potato breeding. Cultivo y fusion de protoplastos These plants were multiplied in glass culture tubes for further analysis.
Production of somatic hybrids by electrofusion in Solanum. Fuxin analysis The numbers of chromosome of S. This value is different from that obtained by Mattheij et al.
Prior to fusion, protoplasts from S. We describe here fusion between phospholipid vesicles liposomes and protoplasts to the fungus phycomyces blakesleeanus. Fusion de protoplastos pdf Microcalli were transferred onto MS medium to induce shoot regeneration under indirect light conditions. Plant material and protoplast isolation. A slot-blot hybridization method for screening somatic hybrids.
The numbers of chromosome of S. Gels were analyzed and photographed under UV light. Somatic hybrids between potato Solanum tuberosum and wild species Solanum vernei exhibit a recombination in the plastome.
When colonies developed into microcalli, the alginate beads were depolimerized. After rpotoplastos fusion and during plant regeneration it frequently happens that genetic changes such as variations in chromosome number occur.
Interspecific hybridization between the cultivated protopllastos Solanum tuberosum subspecies tuberosum L. Root tips were stained by lacto-propionic orcein. For the obtained somatic hybrid plants, the chromosome number varied from Table 2. Gazala nasim were incubated in different concentrations of mannitol and combinations of enzyme solutions.
Nineteen plants fksin regenerated from calli Fig. Successful application of the fusion technique demands a protocol for plant regeneration from protoplasts. Plant Cell Rep 3: The origin of this chromosome instability has been studied elsewhere in potato Kumar, Irregular meiosis in a somatic hybrid between S.
For this analysis, DNA extraction was performed according to Doyle and Doyle with some modifications. Related Articles
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