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Production and analysis of intraspecific somatic hybrids of potato (Solanum tuberosum L.)
Department of Physiological Botany, Uppsala University, Uppsala, Sweden.ORCID iD: 0000-0003-0553-7918
Department of Physiological Botany, Uppsala University, Uppsala, Sweden.
Department of Physiological Botany, Uppsala University, Uppsala, Sweden.
1991 (English)In: Plant Science, ISSN 0168-9452, Vol. 75, no 1, 107-115 p.Article in journal (Refereed) Published
Abstract [en]

Photoplasts of two dihaploid lines of potato were fused to produce a large number of intraspecific somatic hybrid plants among which plants of the expected tetraploid level might be found. Fusion frequencies up to 12% (mean 7%) were observed using a revised polyethylene glycol fusion protocol. Fusion products were identified by the dual fluorescence emission from the chloroplasts in mesophyll protoplasts (red) and from the fluorescein diacetate stain in light and norflurazon bleached protoplasts (yellow-green). Hybrid cells were isolated 2–3 days after fusion and cultured at a cell density of 2000 cells/ml. From a total of 1363 isolated putative hybrid cells, 258 divided to form calli. Plants were regenerated from 166 of these. Isozyme analysis confirmed the hybrid nature in 57 of 58 analysed plants. Ploidy was determined in 51 plants; 12% were tetraploid, 41% hexaploid, 12% octoploid and 35% were mixoploid. Expression of isocitrate dehydrogenase isozymes indicated that the majority of the hexaploid hybrids contained 2 genomes of the bleached parent and one genome of the mesophyll parent. This study shows that tetraploid somatic hybrid potato plants can be obtained by the fusion and selection method presented.

Place, publisher, year, edition, pages
Shannon: Elsevier, 1991. Vol. 75, no 1, 107-115 p.
Keyword [en]
Solanum tuberosum; protoplast fusion, micro manipulation, somatic hybrids, isozyme analysis, DNA analysis by flow cytometry
National Category
Botany
Identifiers
URN: urn:nbn:se:hh:diva-30827DOI: 10.1016/0168-9452(91)90014-YISI: A1991FM35300013Scopus ID: 2-s2.0-0347727759OAI: oai:DiVA.org:hh-30827DiVA: diva2:925851
Available from: 2016-05-03 Created: 2016-05-03 Last updated: 2016-05-03Bibliographically approved
In thesis
1. Production and characterization of intraspecific somatic hybrids of potato (Solanum tuberosum L.)
Open this publication in new window or tab >>Production and characterization of intraspecific somatic hybrids of potato (Solanum tuberosum L.)
1991 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

A culture protocol has been developed for mesophyll protoplasts isolated from various d¡haploid clones of potato. A large number of call¡ was obtained after serial dilution of the cultures w¡th a suitable amount and type of medium at different stages of cell colony development.

A polyelhylene glycol fusion procedure was developed that yielded up to 12 % heterokaryons. Using this fusion protocol hybrid cells have been manually lsolated, cultured and regenerated into plants. Fus¡on products were identified 2-3 days after fuslon by the dual fluorescence emission from the chloroplasts in mesophyll protoplasts (red) and the fluorescein diacetate stain in l¡ght and norflurazon bleached protoplasts (yellow-green). This fusion and select¡on strategy leads to the recovery of almost 100 % hybr¡d plants as established by isozyme analysls.

Cytolog¡cal analysis of protoplast-derived plants and somatic hybrid plants revealed genetic changes as a consequence of protoplast culture and protoplast fus¡on. Twenty-three tetraplo¡d somatic hybrid plants were obta¡ned from 6 different calli and 9 of these were euploid.

Morphological assessments of somatic hybr¡d plants of different ploidy levels demonstrated that tetraploid as well as several hexaploid somatic hybrids showed an increased vigour as compared to the parental plants. Most tetraploid somat¡c hybrids had a similar appearance although not all euploid plants were identical. Loss of vigour was evident in all mixoploid and octoplo¡d plants. These were stunted, weak and had an abnormal leaf morphology.

The plastid type in hybrid plants was determined by RFLP analysis. All analysed hybrids had a cpDNA restrict¡on fragment pattern ¡dent¡cal to one of the parents wh¡ch contained either S. tuberosum or S. stoloniferum cpDNA. A slight preference for S. tuberosum plastids was observed in hybild plants. No influence on tho assortment of chloroplast by the norflurazon bleaching could be detected.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 1991. 44 p.
Series
Comprehensive summaries of Uppsala dissertations from the Faculty of Science, ISSN 0282-7468 ; 323
National Category
Plant Biotechnology
Identifiers
urn:nbn:se:hh:diva-23705 (URN)91-554-2748-0 (ISBN)
Public defence
1991-05-16, Uppsala, 10:00
Note

Financial support was given by the Swedish Council for Forestry and Agricultural Research.

Available from: 2010-02-23 Created: 2013-10-02 Last updated: 2016-05-03Bibliographically approved

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