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Factors promoting sustained divisions of mesophyll protoplasts isolated from dihaploid clones of potato (Solanum tuberosum L.) and a cytological analysis of regenerated plants
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.
Department of Physiological Botany, Uppsala University, Uppsala, Sweden.
1991 (English)In: Plant Cell Tissue and Organ Culture, ISSN 0167-6857, E-ISSN 1573-5044, Vol. 27, no 3, 257-265 p.Article in journal (Refereed) Published
Abstract [en]

A culture protocol has been developed for mesophyll protoplasts isolated from various dihaploid clones of potato. A special effort was made to promote the growth of initially dividing cells to form cell colonies and calli. An increase in plating efficiency in 3 different dihaploid clones and one doubled dihaploid clone was obtained after serial dilution of cultures with a suitable amount and type of medium at different stages of cell colony development. Plating on a refined semi-solid medium after 14 days of culture further improved both the yield and the quality of calli obtained. The refined plating medium also enhanced shoot regeneration ability from 67 to 90% in one of the dihaploid clones (67:9). The refined culture protocol could also be used without causing a decrease in plating efficiency at a low population density adjusted after 3 days of culture. The ploidy level of plants regenerated from dihaploid protoplasts were determined by chromosome counting and DNA analysis by flow cytometry. Most of the plants were aneuploid or tetraploid although, some dihaploid plants were obtained after protoplast culture of 2 dihaploid clones derived from the same cultivar (cv. Stina). 

Place, publisher, year, edition, pages
Dordrecht: Springer Netherlands, 1991. Vol. 27, no 3, 257-265 p.
Keyword [en]
cytological analysis, dihaploid potato, protoplast culture, shoot regeneration, sustained division
National Category
Plant Biotechnology
Identifiers
URN: urn:nbn:se:hh:diva-30777ISI: A1991GV57100004OAI: oai:DiVA.org:hh-30777DiVA: diva2:921585
Available from: 2016-04-20 Created: 2016-04-20 Last updated: 2017-11-30Bibliographically 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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Waara, Sylvia

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