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Rose protoplast isolation and culture and heterokaryon selection by immobilization in extra thin alginate film.

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Title Rose protoplast isolation and culture and heterokaryon selection by immobilization in extra thin alginate film.
 
Creator Pati, P K
Sharma, Madhu
Ahuja, Paramvir Singh
 
Subject Plant sciences
 
Description Somatic hybridization has been identified as one method for
the genetic improvement of roses. The success of somatic hybridization programmes relies to a great extent upon efficient protoplast isolation and culture and selection of heterokaryons. This paper reports the isolation of
rose cell suspension protoplasts by direct sucrose flotation and demonstrates their culture using extra thin alginate film. A comparative assessment of the efficiency of conventional culture techniques versus those
with extra thin alginate film or thin alginate layer is also presented. A very high plating efficiency (80%) was obtained using thin alginate layer or extra thin alginate film techniques with improved media formulations.
Protoplasts of Rosa damascena and R. bourboniana were fused by using polyethylene glycol as fusogen and later immobilized in the thin layer of alginate. The fused protoplasts were tracked on the basis of differential
fluorescent staining, and the hybridity of heterokaryons following their development to callus was confirmed by molecular characterization. This novel selection strategy has general applicability and is faster and simpler
to perform during somatic hybridization experiments.
 
Publisher Springer Wien , Sachsenplatz 4-6, PO Box 89, A-1201 Wien, Austria .
 
Date 2008
 
Type Article
PeerReviewed
 
Format application/pdf
 
Identifier http://ihbt.csircentral.net/128/1/2008_Rose.pdf
Pati, P K and Sharma, Madhu and Ahuja, Paramvir Singh (2008) Rose protoplast isolation and culture and heterokaryon selection by immobilization in extra thin alginate film. Protoplasma, 233 (1-2). pp. 165-171. ISSN 0033-183X
 
Relation http://ihbt.csircentral.net/128/