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Somatic embryogenesis and plant regeneration from immature zygotic embryo cultures of mountain ash (Sorbus pohuashanensis)

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Abstract

Plant regeneration through somatic embryogenesis was achieved using immature zygotic embryos (ZE) of Sorbus pohuashanensis as explants. Over 50% of immature ZEs from immature seed collected at 30 days after pollination produced direct somatic embryos (SEs) on Murashige and Skoog (MS) medium supplemented with 0–0.44 μM 6-benzyladenine (BA) in combination with 5.73 μM naphthaleneacetic acid (NAA) or with 0.91–2.26 μM 2,4-dichlorophenoxyacetic acid (2,4-D) alone. Fourteen to 23 SEs per explant were regenerated on MS medium supplemented with BA 0.44 μM in combination with NAA 5.73 μM. SE formation decreased when sucrose concentrations were higher than 40 g L−1. Repetitive embryogenesis occurred following culture on solid MS medium containing 12 μM abscisic acid, 75 g L−1 polyethylene glycol, and 20 g L−1 sucrose at 25 ± 1°C under a 16-h photoperiod with a light intensity of 40 μmol m−2 s−1. Over 40% of the mature SEs germinated on solid MS medium under light condition described previously. Up to 40% of the regenerated plantlets were successfully acclimatized under greenhouse conditions. Plantlets derived from SEs grew vigorously with similar morphology as those germinated from ZEs. Histological studies of explants at various developmental stages of somatic embryogenesis revealed that SEs passed through globular, heart, torpedo, and mature stages. Similar to ZE suspensors, similar structures of SE degenerated in later stages of embryo development. ZE and SE are a effective means of regenerating tissue culture plantlets for S. pohuashanesis.

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Abbreviations

½ MS:

Half-strength of MS macroelements

2,4-D:

2,4-dichlorophenoxyacetic acid

ABA:

Abscisic acid

BA:

6-benzyladenine

CH:

Casein hydrolysate

IAA:

Indole-3-acetic acid

IBA:

Indole-3-butyric acid

MS:

Murashige and Skoog (1962) medium

NAA:

Naphthaleneacetic acid

PEG:

Polyethylene glycol

PGR(s):

Plant growth regulator(s)

SE(s):

Somatic embryo(s)

SH:

Schenck and Hildebrandt (1972)

SSE(s):

Secondary somatic embryo(s)

TDZ:

Thidiazuron

WPM:

Woody plant medium (1980)

ZE(s):

Zygotic embryo(s)

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Acknowledgments

This research was supported by the China Postdoctoral Science Foundation (20110491015), Fundamental Research Funds for the Central Universities of China (DL10BA04), and the Postdoctoral Science Foundation of Heilongjiang Province of China (LBH-10284). We thank for the kind help of Dr. Daniel Struve, professor from Ohio State University, USA, for reviewing the revised copy of this manuscript, and thank all the colleagues in our lab for constructive discussion and technical support.

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  1. State Key Laboratory of Forest Genetics and Breeding, Northeast Forestry University, Harbin, 150040, China

    Ling Yang, Yuhua Li & Hailong Shen

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  1. Ling Yang
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  2. Yuhua Li
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  3. Hailong Shen
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Correspondence to Hailong Shen.

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Yang, L., Li, Y. & Shen, H. Somatic embryogenesis and plant regeneration from immature zygotic embryo cultures of mountain ash (Sorbus pohuashanensis). Plant Cell Tiss Organ Cult 109, 547–556 (2012). https://doi.org/10.1007/s11240-012-0121-1

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