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Efficient procedure for grapevine embryogenic suspension establishment and plant regeneration: role of conditioned medium for cell proliferation

  • Cell Biology and Morphogenesis
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Abstract

An efficient system for the establishment and multiplication of highly prolific embryogenic cell cultures of grapevine (Vitis sp.) was developed. Using anther-derived pro-embryogenic masses as starting material, cell suspensions of different grapevine cultivars (Tempranillo, Cabernet-Sauvignon) and rootstocks (Kober 125 AA, Kober 5 BB, 110 Richter) were initiated in liquid medium containing NOA (1.0 mg l-1) and BAP (0.25 mg l-1) as growth regulators. Conditioned medium was recovered and utilised for establishing new, highly totipotent cell cultures. The suspensions obtained, showed embryogenic competence resulting in somatic embryo induction and subsequent plant regeneration. In this study, a simplified establishment procedure for grapevine embryogenic cell suspension allowing the fast multiplication of embryogenic material is described. Evidence for the promoting effect of the protein fraction derived from conditioned medium, on cell proliferation was found. In bioassays, addition of ß-d-GlcY affect cell proliferation suggesting that arabinogalactan proteins are required for growth processes in grapevine cell cultures.

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Abbreviations

AGPs:

Arabinogalactan-proteins

BAP:

6-benzylaminopurine

ß-D-Glc-Y:

ß-d-Glucosyl Yariv phenylglycoside

CEPF:

Concentrated extra-cellular protein fraction

2,4-D:

2,4-Dichlorophenoxyacetic acid

EP:

Extra-cellular protein

NOA:

Naphthoxyacetic acid

MES:

2-(N-Morpholino) ethanesulfonic acid

FM:

Fresh medium

CM:

Conditioned medium

PEM:

Pro-embryogenic masses

PCV:

Packed cell volume

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Authors and Affiliations

  1. Laboratoire de Physiologie Moléculaire de la Vigne, Technopôle de Borj Cédria, BP 95, Hammam-Lif, 2050, Tunisia

    A. Ben Amar & A. Ghorbel

  2. Faculté des Sciences de Tunis, Laboratoire de Morphogenèse & Biotechnologie végétale, Campus El Manar, Tunis, 2092, Tunisia

    A. Ben Amar & S. Bouzid

  3. RLP AgroScience GmbH, AlPlanta, Institute for Plant Research, Breitenweg 71, 67435, Neustadt/Weinstraße, Germany

    A. Ben Amar, P. Cobanov, K. Boonrod, G. Krczal & G. M. Reustle

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  1. A. Ben Amar
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  2. P. Cobanov
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  3. K. Boonrod
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  4. G. Krczal
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  5. S. Bouzid
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  6. A. Ghorbel
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  7. G. M. Reustle
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Correspondence to G. M. Reustle.

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Communicated by R. Reski.

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Ben Amar, A., Cobanov, P., Boonrod, K. et al. Efficient procedure for grapevine embryogenic suspension establishment and plant regeneration: role of conditioned medium for cell proliferation. Plant Cell Rep 26, 1439–1447 (2007). https://doi.org/10.1007/s00299-007-0341-8

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  • DOI: https://doi.org/10.1007/s00299-007-0341-8

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