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Gynogenesis in gentians (Gentiana triflora, G. scabra): production of haploids and doubled haploids

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Abstract

Gynogenesis was investigated on gentian (Gentiana triflora, G. scabra and their hybrids), which is an important ornamental flower. When unfertilized ovules were cultured in 1/2 NLN medium containing a high concentration of sucrose (100 g/l), embryo-like structures (ELS) were induced. Although genotypic variation was observed in ELS induction, all four genotypes produced ELSs ranging from 0.93 to 0.04 ELSs per flower bud. The ovules collected from flower buds of later stages (just before anthesis or flower anthesis) tended to exhibit higher response. The dark culture condition produced more than four times as many ELSs than in 16-h light condition. A significant number of plantlets were directly regenerated from ELSs on MS regeneration medium. The ploidy levels of 179 regenerated plants were determined by flow cytometry, revealing that the majority of them were diploid (55.9%) and haploid (31.3%). When a total of 54 diploid plants were examined by molecular genetic markers, 52 (96.3%) were considered as doubled haploids (DHs). This is the first report showing successful gynogenesis in gentian. The production of haploids and DHs by unfertilized ovule culture opens a novel prospect in gentian F1 hybrid breeding.

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Abbreviations

ELS:

Embryo-like structure

DH:

Doubled haploid

MS:

Murashige and Skoog basal salt and vitamins

GA3 :

Gibberellic acid

FLS:

Flavonol synthase

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Acknowledgments

This work was supported in part by “Research and Development Projects for Application in Promoting New Policy of Agriculture, Forestry and Fisheries” from the Ministry of Agriculture, Forestry and Fisheries of Japan.

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Authors and Affiliations

  1. Faculty of Agriculture, Iwate University, Morioka, 020-8550, Japan

    Hisako Doi, Shuji Yokoi, Ken-ich Tsutsumi & Yoshihito Takahata

  2. Hachimantai City Floricultural Research and Development Center, Hachimantai, 028-7592, Japan

    Takashi Hikage

  3. Iwate Biotechnology Center, Kitakami, 024-0003, Japan

    Masahiro Nishihara

Authors
  1. Hisako Doi
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  2. Shuji Yokoi
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  3. Takashi Hikage
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  4. Masahiro Nishihara
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  5. Ken-ich Tsutsumi
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  6. Yoshihito Takahata
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Correspondence to Yoshihito Takahata.

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Communicated by R. Rose.

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Doi, H., Yokoi, S., Hikage, T. et al. Gynogenesis in gentians (Gentiana triflora, G. scabra): production of haploids and doubled haploids. Plant Cell Rep 30, 1099–1106 (2011). https://doi.org/10.1007/s00299-011-1017-y

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  • DOI: https://doi.org/10.1007/s00299-011-1017-y

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