1987) or gamma-irradiation (Ralushnyak et al. Iodoacetamide (IOA) treatment of one partner (OziasAkins et al. Methods have been reported for the selection of parasexual hybrids: (1) prefusion treatments of protoplasts, like 1991) have also been utilized as fusion partners. InĪddition, two types of leaf protoplasts (Wijbrandi et al.ġ990), or leaf and irradiated leaf protoplasts (RalushnyakĮt al. Method employed (O’Connell and Hanson 1985, 1986). Fusion between leaf and suspension-culture-derived protoplasts has been the major To date, somatic hybridization has been reported with different materials, fusion methods, and selection methods for To transfer desirable agronomic traits from wild to cultivated species (Lefrancois et al. Recombinations of nuclear and cytoplasmic genomes, and Somatic hybridization by protoplast fusion provides the tool to increase genetic variation through (Chen and Imanishi 1991) have been used to obtain interspecific hybrids between L. Embryo culture (Smith 1944 Ĭhen and Adachi 1992, 1996) as well as ovule culture peruvianum is only distantly related to the cultivated tomato L. Gakuen Kibanadai Nishi 1-1, Miyazaki 889-2192, JapanĮ-mail: to vario us diseases and pests (Rick and Yoder 1988). SyonoĪpplied Genetics and Biotechnology Division,įaculty of Agriculture, Miyazaki University, peruvianum is considered to be a source of agronomic traits, such as resisCommunicated by K. It is well known as a model plant forĬytological and molecular research. Tomato, Lycopersicon esculentum, is an economically important crop species. Observations indicated that the hybrids were tetraploid, Were intermediate to their donor partners, and chromosome TheĮxcised torpedo-stage embryos could be propagated on a Torpedo stages, and finally formed complete plantlets. The somatic embryos were derivedįrom greenish calli that formed from the fusion mixtures,ĭeveloped progressively through the globular, heart, and Of a culture medium which only allowed cotyledon protoplasts to regenerate. The hybrids were selected by a multiple-step selection procedure relying on the different colors of the fusion partners, IOA treatment of cotyledon protoplasts, and the use Kyoryokutoko treated with iodoacetamide (IOA) and suspension-culture-derived protoplasts Glycol fusion of cotyledon protoplasts of Lycopersicon esculentum Mill. Received: 24 July 1997 / Revision received: 4 November 1997 / Accepted: 2 December 1997Ībstract Somatic hybrids were obtained by polyethylene peruvianum-complex: somatic embryogenesis,
In Vitro Cellular & Developmental Biology - Plant Springer Journals Protoplast fusion between Lycopersicon esculentumĪnd L. × leptoeuropaea gave rise to cell colonies and betweenL. × laricina protoplasts and protoplasts ofL. Heterofusion frequencies of approximately 18% and 6% in the presence of PEG and an electric field, respectively, were attained. The fusion products could be identified by dual fluorescence several h after the onset of fusion. The progress of fusion was readily observed, taking only minutes under the experimental conditions. The fusion of mixed parental protoplasts that were internally labeled was carried out either in the presence of a 20% polyethylene glycol (PEG) solution or in an electric field. Of a number of dyes tested, only rhodamine B hexyl ester chloride (R6) and 3,3′-dihexylox-carbocyanine iodide (DiOC6) stained the protoplasts in a consistent and uniform fashion. A method of unambiguous labeling of parental protoplasts prior to fusion was developed using vital fluorescent dyes. laricina line L2 was used in fusion experiments with eitherL. Larix was chosen for the study on interspecies protoplast fusion due to its ability to regenerate plants from protoplasts derived from embryogenic cultures.L. Pattanavibool, Rungnapar Klimaszewska, Krystyna Aderkas, Patrick Interspecies protoplast fusion inLarix: Comparison of electric and chemical methods Interspecies protoplast fusion inLarix: Comparison of electric and chemical methods
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