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Shoot organogenesis in leaf explants of Hydrangea macrophylla ‘Hyd1’ and assessing genetic stability of regenerants using ISSR markers

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Abstract

For the first time, an in vitro regeneration protocol of Hydrangea macrophylla ‘Hyd1’ was developed. Effects of different plant growth regulators (PGRs) on shoot regeneration were investigated jointly with selecting optimal basal media and cefotaxime concentrations. The highest frequency of shoot organogenesis (100%) and mean number of shoots per explant (2.7) were found on Gamborg B5 basal medium supplemented with 2.25 mg/l 6-benzyladenine (BA), 0.1 mg/l Indole-3-butyric acid (IBA), 100 mg/l cefotaxime and 30 g/l sucrose solidified by 7 g/l agar. Regenerated shoots were rooted by culturing on perlite plus half strength liquid B5 basal medium with 0.5 mg/l NAA. Rooted plantlets were transplanted to the greenhouse with 100% survival rate. Genetic stability of 32 plantlets (one mother plant and 31 regenerants) was assessed by 44 ISSR markers. Out of 44 ISSR markers, ten markers produced clear, reproducible bands with a mean of 5.9 bands per marker. The in vitro regeneration protocol is potentially useful for the genetic transformation of Hydrangea macrophylla ‘Hyd1’.

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Fig. 1
Fig. 2

Abbreviations

B5:

Gamborg B5 medium

BA:

6-benzyladenine

IAA:

Indoleacetic acid

IBA:

Indole-3-butyric acid

ISSR:

Inter simple sequence repeat

MS:

Murashige and Skoog medium

NAA:

α-Naphthalene acetic acid

PCR:

Polymerase chain reaction

RAPD:

Random amplified polymorphic DNA

WPM:

Woody plant medium

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Acknowledgments

The work was supported by Agriom B.V. (the Netherlands), and the Dutch EVD Asia Facility for China program (project no. AF05CH01).

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Authors and Affiliations

  1. Key Laboratory for Biology of Horticultural Plants, Ministry of Education, College of Horticulture and Forestry Sciences, Huazhong Agricultural University, 430070, Wuhan, People’s Republic of China

    Feng Liu, Li–Li Huang, Yang-Li Li & Cai-Yun Wang

  2. Agriom B. V., Agriom, Achterweg 58A, 1424, PR De Kwakel, The Netherlands

    Poula Reinhoud

  3. Plant Research International, Wageningen UR, Postbus 619, 6700 AP, Wageningen, The Netherlands

    Maarten A. Jongsma

Authors
  1. Feng Liu
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  2. Li–Li Huang
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  3. Yang-Li Li
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  4. Poula Reinhoud
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  5. Maarten A. Jongsma
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  6. Cai-Yun Wang
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Correspondence to Cai-Yun Wang.

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Liu, F., Huang, L., Li, YL. et al. Shoot organogenesis in leaf explants of Hydrangea macrophylla ‘Hyd1’ and assessing genetic stability of regenerants using ISSR markers. Plant Cell Tiss Organ Cult 104, 111–117 (2011). https://doi.org/10.1007/s11240-010-9797-2

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  • DOI: https://doi.org/10.1007/s11240-010-9797-2

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