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From callus to embryo: a proteomic view on the development and maturation of somatic embryos in Cyclamen persicum

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Abstract

In this study, the proteome structures following the pathway in somatic embryogenesis of Cyclamen persicum were analysed via high-resolution 2D-SDS-PAGE with two objectives: (1) to identify the significant physiological processes during somatic embryogenesis in Cyclamen and (2) to improve the maturation of somatic embryos. Therefore, the effects of maturation-promoting plant growth regulator abscisic acid (ABA) and high sucrose levels on torpedo-shaped embryos were investigated. In total, 108 proteins of differential abundance were identified using a combination of tandem mass spectrometry and a digital proteome reference map. In callus, enzymes related to energy supply were especially distinct, most likely due to energy demand caused by fast growth and cell division. The switch from callus to globular embryo as well as from globular to torpedo-shaped embryo was associated with controlled proteolysis via the ubiquitin-26S proteasome pathway. Storage compound accumulation was first detected 21 days after transfer to plant growth regulator (PGR)-free medium in early torpedo-shaped embryos. Increase in abundance of auxin-amidohydrolase during embryogenesis suggests a possible increase in auxin release in the late embryo stages of Cyclamen. A development-specific isoelectric point switch of catalases has been reported for the first time for somatic embryogenesis. Several proteins were identified to represent markers for the different developmental stages analysed. High sucrose levels and ABA treatment promoted the accumulation of storage compounds in torpedo-shaped embryos. Additionally, proteins of the primary metabolic pathways were decreased in the proteomes of ABA-treated embryos. Thus, ABA and high sucrose concentration in the culture medium improved maturation and consequently the quality of somatic embryos in C. persicum.

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Abbreviations

2D:

Two-dimensional

2iP:

6-(γ-γ-Dimethylallylamino) purine

2,4-D:

2,4-Dichlorophenoxyacetic acid

ABA:

Abscisic acid

EDTA:

Ethylenediaminetetraacetic acid

GAPDH:

Glyceraldehyde-3-phosphate dehydrogenase

HSP:

Heat shock protein

IEF:

Isoelectric focusing

IPG:

Immobilized pH gradients

LEA:

Late embryogenesis abundant proteins

MS:

Mass spectrometry

PGR:

Plant growth regulator

RNase:

Endoribonuclease

SDS-PAGE:

Sodium dodecyl sulphate polyacrylamide gel electrophoresis

SERK:

Somatic embryogenesis receptor like kinase

SOD:

Superoxide dismutase

TCA:

Tricarboxylic acid

VDA channel:

Voltage-dependent anion channel

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Acknowledgments

The authors would like to thank Dr. Frank Colditz, Institute of Plant Genetics, Leibniz Universitaet Hannover, for proof reading and critical discussion, Jenniffer Mwangi for proof reading, Michael Senkler for computer-related assistance and the DFG (German research foundation) for financial support.

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Authors and Affiliations

  1. Institute of Plant Genetics, Leibniz Universität Hannover, Herrenhäuser Str. 2, 30419, Hannover, Germany

    Christina Rode & Hans-Peter Braun

  2. Institute of Floriculture and Woody Plant Science, Leibniz Universität Hannover, Herrenhäuser Str. 2, 30419, Hannover, Germany

    Kathrin Lindhorst & Traud Winkelmann

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  1. Christina Rode
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  2. Kathrin Lindhorst
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  4. Traud Winkelmann
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Correspondence to Traud Winkelmann.

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Rode, C., Lindhorst, K., Braun, HP. et al. From callus to embryo: a proteomic view on the development and maturation of somatic embryos in Cyclamen persicum . Planta 235, 995–1011 (2012). https://doi.org/10.1007/s00425-011-1554-1

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