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Method Article
Philippe Vergne
Laboratoire Reproduction et Développement des Plantes, Univ Lyon, ENS de Lyon, UCB Lyon 1, CNRS, INRA, F-69364 Lyon, France.
Corresponding Author
Mohammed Bendahmane
Laboratoire Reproduction et Développement des Plantes, Univ Lyon, ENS de Lyon, UCB Lyon 1, CNRS, INRA, F-69364 Lyon, France.
Corresponding Author
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Like many economically important plants, roses exhibit high heterozygosity levels. Here, we report the development of a protocol that allows rose microspores to switch from gametophyte to sporophyte development. Using a combination of starvation medium fine-tuning, cold stress and hormonal treatments we were able to induce microspores to initiate divisions and form cell clusters from which embryogenic and proliferating calli could be obtained. DNA genotyping showed that the genome of these calli was homozygous, demonstrating the loss of heterozygosity in the rose. Homozygous calli maintained their embryogenic capacity through several subcultures. Plantlets with normal morphological phenotype could be regenerated from the homozygous embryos. To the best of our knowledge, this is the first demonstration of the production of a homozygous rose plantlet. The use of such approach helps reduce genome complexity to obtain high quality genome assembly and also opens possibilities to implement haplomethods in rose genetics and breeding.
Keywords
Rose, microspore, androgenesis, callus, embryogenic, homozygous, plant regeneration
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The authors declare no competing financial interests
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An open repository of community-contributed protocols sponsored by Nature Research.
Learn more about Protocol Exchange
Method Article
Philippe Vergne
Laboratoire Reproduction et Développement des Plantes, Univ Lyon, ENS de Lyon, UCB Lyon 1, CNRS, INRA, F-69364 Lyon, France.
Corresponding Author
Mohammed Bendahmane
Laboratoire Reproduction et Développement des Plantes, Univ Lyon, ENS de Lyon, UCB Lyon 1, CNRS, INRA, F-69364 Lyon, France.
Corresponding Author
Version History
CURRENT STATUS: Posted
Version 1
Posted 22 May, 2018
No community comments so far
Metrics
Comments: 0
HTML Views: ...
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Like many economically important plants, roses exhibit high heterozygosity levels. Here, we report the development of a protocol that allows rose microspores to switch from gametophyte to sporophyte development. Using a combination of starvation medium fine-tuning, cold stress and hormonal treatments we were able to induce microspores to initiate divisions and form cell clusters from which embryogenic and proliferating calli could be obtained. DNA genotyping showed that the genome of these calli was homozygous, demonstrating the loss of heterozygosity in the rose. Homozygous calli maintained their embryogenic capacity through several subcultures. Plantlets with normal morphological phenotype could be regenerated from the homozygous embryos. To the best of our knowledge, this is the first demonstration of the production of a homozygous rose plantlet. The use of such approach helps reduce genome complexity to obtain high quality genome assembly and also opens possibilities to implement haplomethods in rose genetics and breeding.
Figure 1
Figure 2
The authors declare no competing financial interests
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