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Embryogenesis and doubled haploid production from anther culture in gentian (Gentiana triflora)

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Abstract

The overall goal of this study is to develop an anther culture system to produce doubled haploid (DH) lines of gentian (Gentiana triflora), an ornamental flowering plant, for use in an F1 hybrid breeding program. Embryogenesis was induced from anther cultures incubated on half-strength modified Lichter (NLN) medium containing a high concentration of sucrose (130 g/l) and subjected to heat shock treatment. Among the various parameters investigated, anthers collected from buds 9–12 mm in length induced the highest frequency of androgenesis. Moreover, among three genotypes tested, cvs. Ashiro-no-Aki and Ashiro-no-Natsu produced 21.3 and 3.7 embryos per 100 anthers, respectively, whereas, cv. Lovely-Ashiro failed to produce embryos. Among a total of 427 embryos transferred to a regeneration medium consisting of Murashige and Skoog (MS) medium, 138 plants were regenerated. The ploidy levels of regenerants were determined by flow cytometry and chromosome counts, revealing the presence of 5% haploids, 25% diploids, and 70% triploids. Inter simple sequence repeat (ISSR) analysis using the 6PS line obtained following self-pollination of the diploid plant obtained from anther culture confirmed that the diploid plant was indeed a DH.

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Acknowledgments

We greatly appreciate Dr. J. Soejima of the Apple Research Station, National Institute of Fruit Tree Science, Japan, for his help in providing the PA-I flow cytometer (Partec).

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Authors and Affiliations

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

    Hisako Doi & Yoshihito Takahata

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

    Ryo Takahashi & Takashi Hikage

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  1. Hisako Doi
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  2. Ryo Takahashi
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  3. Takashi Hikage
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  4. Yoshihito Takahata
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Correspondence to Yoshihito Takahata.

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Doi, H., Takahashi, R., Hikage, T. et al. Embryogenesis and doubled haploid production from anther culture in gentian (Gentiana triflora). Plant Cell Tiss Organ Cult 102, 27–33 (2010). https://doi.org/10.1007/s11240-010-9700-1

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  • DOI: https://doi.org/10.1007/s11240-010-9700-1

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