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Plant regeneration with maintenance of the endosperm ploidy level by endosperm culture in Lonicera caerulea var. emphyllocalyx

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Abstract

An endosperm culture of Haskap (Lonicera caerulea var. emphyllocalyx) was established to develop polyploid plants and investigate the regeneration ability of the endosperm. Based on histological analysis of embryo and endosperm development, endosperms at the globular to early torpedo-stages of developing embryos were used to initiate an endosperm culture. Formation of shoot primordia was observed on Murashige and Skoog (MS) medium (Physiol Plant 15:473–497, 1962) containing benzyladenine and indole-3-butyric acid. Shoot primordium formation was confirmed in some genotypes with regeneration frequencies ranging between 1.9 and 10.0%. These proliferated on ½ MS medium containing 2.89 μM gibberellic acid (GA3), and then elongated and rooted on MS medium containing 0.44 μM BA and 2.89 μM GA3. These shoots developed into plantlets on ½ MS medium. Plantlets maintained ploidy of the endosperm following flow cytometric analysis, thus confirming that these were derived from the endosperm. These results indicated that endosperms were capable of regeneration.

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Abbreviations

BA:

6-Benzyladenine

CH:

Casein hydrolysate

DAP:

Days after pollination

DAPI:

4′,6-Diamidino-2-phenylindole

GA3 :

Gibberellic acid

IBA:

Indole-3-butyric acid

MS:

Murashige and Skoog (1962) medium

NAA:

1-Naphthaleneacetic acid

PGR:

Plant growth regulator

TDZ:

Thidiazuron

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Acknowledgments

Authors thank Dr. K. Sahara for valuable suggestions on chromosome observation. We are grateful to Professor H. Nakashima for the supports of plant managements. We also gratefully acknowledge H. Hori, H. Tamura, M. Ikuta, H. Nakano and S. Takamushi for the technical assistances on the University farms. This work was supported in part by grants from Inamori Foundation, Takeda Scientific foundation, and a Grant-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports Science and Technology, Japan.

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Authors and Affiliations

  1. Course in Agro-Ecosystem Science, Division of Biosphere Science, Graduate School of Environmental Science, Hokkaido University, Kita 11, Nishi 10, Kita-Ku, Sapporo, 060-0811, Japan

    Tomomi Miyashita & Yoichiro Hoshino

  2. Field Science Center for Northern Biosphere, Hokkaido University, Kita 11, Nishi 10, Kita-Ku, Sapporo, 060-0811, Japan

    Takafumi Ohashi, Hajime Araki & Yoichiro Hoshino

  3. Department of Applied Bioscience, Graduate School of Agriculture, Hokkaido University, Kita 9, Nishi 9, Kita-Ku, Sapporo, 060-8589, Japan

    Fukashi Shibata

  4. Division of Innovative Research, Creative Research Initiative ‘Sousei’ (CRIS), Hokkaido University, Kita 21, Nishi 10, Kita-Ku, Sapporo, 001-0021, Japan

    Yoichiro Hoshino

Authors
  1. Tomomi Miyashita
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  2. Takafumi Ohashi
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  3. Fukashi Shibata
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  4. Hajime Araki
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  5. Yoichiro Hoshino
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Correspondence to Yoichiro Hoshino.

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Miyashita, T., Ohashi, T., Shibata, F. et al. Plant regeneration with maintenance of the endosperm ploidy level by endosperm culture in Lonicera caerulea var. emphyllocalyx . Plant Cell Tiss Organ Cult 98, 291–301 (2009). https://doi.org/10.1007/s11240-009-9562-6

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  • DOI: https://doi.org/10.1007/s11240-009-9562-6

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