Incorporation of hygromycin resistance in Brassica nigra and its transfer to B. napus through asymmetric protoplast fusion

1989 ◽  
Vol 78 (2) ◽  
pp. 194-200 ◽  
Author(s):  
M. D. Sacristán ◽  
M. Gerdemann-Knörck ◽  
O. Schieder
Keyword(s):  
Protoplast Fusion ◽  
Brassica Nigra ◽  
Hygromycin Resistance ◽  
Asymmetric Protoplast Fusion

scholarly journals An asymmetric protoplast fusion and screening method for generating celeriac cybrids

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Silvia Bruznican ◽  
Tom Eeckhaut ◽  
Johan Van Huylenbroeck ◽  
Ellen De Keyser ◽  
Hervé De Clercq ◽  
...  
Keyword(s):  
Protoplast Fusion ◽  
Screening Method ◽  
Low Frequency ◽  
Nuclear Genome ◽  
Hybrid Seed Production ◽  
Male Sterile ◽  
Suspension Cells ◽  
Asymmetric Protoplast Fusion ◽  
Pooled Samples ◽  
Hybrid Screening

AbstractCeleriac F1 hybrid seed production is currently complicated due to the instability of cytoplasmic male sterile lines. To develop alternative alloplasmic CMS lines, an asymmetric protoplast fusion and hybrid screening methodology was established. Celeriac suspension cells protoplasts were used as the acceptor and carrot, coriander and white celery mesophyll protoplasts as the donor for protoplast fusion experiments. Acceptor cytoplasmic inheritance was inhibited by iodoacetamide treatment and donor nuclear genome inheritance was prevented by UV exposure. Protoplasts were selectively stained and fused using electroporation and polyethylene glycol, and candidate hybrid shoots were obtained. One chloroplast and three mitochondrial markers that could distinguish acceptor and donors organelles were used to characterize over 600 plants obtained after fusion events, without identifying any cybrid. In order to increase the testing efficiency a high number of micro plantlets were pooled and hence the presence of the carrot specific Atp1 marker in one of the pooled samples was detected. We demonstrated that fusion took place between celeriac and a carrot indicating that the creation of viable hybrids is possible although at a very low frequency. These findings open the path for new cytoplasmic hybridization and the isolation of novel CMS lines of celeriac.

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