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Somatic embryogenesis and fertile green plant regeneration from suspension cell-derived protoplasts of rye (Secale cereale L.)

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Abstract

A method for somatic embryogenesis and fertile green plant regeneration from suspension cell-derived protoplasts of rye (Secale cereale L. cv. Auvinen) was developed. Fast-growing and friable embryogenic calli with a high regeneration capacity were induced from immature rye inflorescences using modified MS medium. These friable embryogenic calli were used for suspension culture initiation in liquid AA medium. A high yield of protoplasts was obtained from suspension cell clumps after 3–5 days of subculture. Isolated protoplasts were cultured in KM8p medium. The frequency of protoplast cell divisions and colony formations in liquid culture medium were similar to those on agarose-solidified medium. Compact embryogenic calli were developed from protoplast-derived microcalli in growth medium mMS. Approximately 7% of the transferred embryogenic calli produced green shoots on N6 regeneration medium. Of 33 green plants, 28 were fertile with normal flowering and seed set. The ratio of green and albino plantlets was 1:4. Rye protoplast-derived green plants showed normal diploid characters as determined by flow cytometer analysis and chromosome counting.

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Fig. 1a–i.
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Fig. 4a–d.

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Abbreviations

2,4 - D :

2,4-Dichorophenoxyacetic acid

FDA :

Fluorescein diacetate

FW :

Fresh weight

GA 3 :

Gibberellic acid

Kinetin :

6-Furfurylaminopurine

IAA :

Indole-3-acetic acid

NAA :

α-Naphthaleneacetic acid

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Acknowledgements

Ms. Marja Piirainen, Mr. Jukka Karhu, Ms. Mervi Ylämäki and the staff of the Laboratory of Plant Physiology and Molecular Biology are thanked for their technical assistance. This work was supported by the Finnish Ministry of Agriculture and Forestry.

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Authors and Affiliations

  1. Laboratory of Plant Physiology and Molecular Biology, Department of Biology, University of Turku, 20014, Turku, Finland

    R. Ma, Y.-D. Guo & S. Pulli

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  1. R. Ma
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  2. Y.-D. Guo
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  3. S. Pulli
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Correspondence to R. Ma.

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Communicated by H. Lörz

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Ma, R., Guo, YD. & Pulli, S. Somatic embryogenesis and fertile green plant regeneration from suspension cell-derived protoplasts of rye (Secale cereale L.). Plant Cell Rep 22, 320–327 (2003). https://doi.org/10.1007/s00299-003-0694-6

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  • DOI: https://doi.org/10.1007/s00299-003-0694-6

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