Protoplast

About: Protoplast is a research topic. Over the lifetime, 5474 publications have been published within this topic receiving 122468 citations.

Production and analysis of plants that are somatic hybrids of barley (Hordeum vulgare L.) and carrot (Daucus carota L.)

Abstract: In order to obtain plants that were somatic hybrids of barley (Hordeum vulgare L.) and carrot (Daucus carota L.), we fused protoplasts that had been isolated from 6-month-old suspension cultures of carrot cells with protoplasts isolated from barley mesophyll by electrofusion. After culture for 1 month at 25°C , the cells were cultured for 5 weeks at 4°C , and were then returned to 25°C for culture on a shoot-inducing medium. Three plants (nos. 1, 2 and 3) were regenerated from the cells. The morphology of the regenerated plants closely resembled that of the parental carrot plants. A cytological analysis of callus cultures induced from these plants indicated that most of the cells had about 24 chromosomes, fewer than the sum of the numbers of parent chromosomes which was 32. Southern hybridization analysis with fragments of the rgp1 gene used as probe showed that the regenerated plants contained both barley and carrot genomic DNA. Chloroplast (ct) and mitochondrial (mt) DNAs were also analyzed with several probes. The ctDNA of the regenerated plants yielded hybridization bands specific for both barley and carrot when one fragment of rice ctDNA was used as probe. Furthermore, the regenerated plants yielded a barley specific band and a novel band with another fragment of rice ct DNA as a probe. One of the regenerated plants (no. 1) yielded a novel pattern of hybridized bands of mt DNA (with an atp6 probe) that was not detected with either of the parents. These results indicated that the regenerated plants were somatic hybrids of barley and carrot and that recombination of both the chloroplast genomes and the mitochondrial genomes might have occurred.

Electric-Field-Stimulated Fusion: Increased Field Stability of Cells Induced by Pronase

Abstract: Zinnia cells and protoplasts showed that the protoplast system can be much better standardized because for protoplast isolation sterile cultured plants can be used. The experiments are continued with the aim (among others) to increase the yield of TE. If this should be the case, the system offers the chance for examining a fundamental phenomenon of cytodifferentiation under aspects of somatic genetics. It will present the advantage to influence the process of differentiation by introduction of foreign genetic material via incorporation or fusion experiments.

Induction of somatic embryogenesis and caulogenesis from cotyledon and leaf protoplast-derived colonies of melon (Cucumis melo L.).

Abstract: A procedure leading to the regeneration of whole plants from protoplasts of melon is described. Protoplasts were isolated from cotyledons and leaves of plants grown in vitro. After 14 days of culture, average viability and division rates were respectively 60% and 30% for the two organs, considering total initial protoplasts plated. The manipulation of the exogenous auxin / cytokinin balance in regeneration media enabled to direct morphogenesis towards somatic embryogenesis (1 mg·l(-1) 2,4-dichlorophenoxyacetic acid and 0.1 mg·l(-1) 6-benzylaminopurine) or caulogenesis (0.5 mg·l(-1) 6-benzylaminopurine and 0.5 mg·l(-1) kinetin). Contrary to division ability, regeneration capacity was genotype-dependent under our conditions, but the two organs expressed similar division and regeneration capacities. Maltose was superior to sucrose for the development of caulogenic nodules into buds. Some plants were transplanted to soil, where they appeared to be fertile and produced seeds.