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Somatic embryogenesis in saffron (Crocus sativus L.). Histological differentiation and implication of some components of the antioxidant enzymatic system

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Abstract

The ontogenetic developmental stages of saffron somatic embryogenesis have been studied and characterized using light microscopy and the biochemical determination of the antioxidant enzymatic system. The embryogenic callus underwent internal segmented divisions with the formation of globular embryos that were attached to the callus surface by a broad multicellular structure. Further development of the embryoids was characterized by the emergence of a shoot apical meristem and cotyledon (monopolar stage) with the subsequent differentiation of a minicorm at the basal part of the somatic embryo (dipolar stage). During the morphological differentiation of the somatic embryos changes in the antioxidant enzymatic system with increased superoxide dismutase (SOD) and catalase (CAT) activities were detected at the initial stages of somatic embryogenesis. The isoforms of SOD, including two Mn-SODs and four Cu, Zn-SODs, were also detected. Although all the isoforms were expressed during the successive stages of somatic embryogenesis, an increase in Mn-SODs and a decrease in Cu, Zn-SODs during the last two stages was observed. Significant changes were also detected in the antioxidant activities ascorbate peroxidase, dehydroascorbic acid reductase and glutathione reductase.

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Abbreviations

APX:

Ascorbate peroxidase

ASC:

Ascorbate

BA:

Benzyladenine

CAT:

Catalase

2,4 D:

2,4 Dichlorophenoxiacetic acid

EDTA:

Ethylenediaminetetraacetic acid

DHA:

Dehydroascorbic acid

DHAR:

Dehydroascorbic acid reductase

GR:

Glutathione reductase

GSSG:

Glutathione oxidized

GSH:

Glutathione reduced

KCN:

Potassium cyanide

MDA:

Malondialdehyde

MDHAR:

Monodehydroascorbate reductase

NAA:

α-Naphtaleneacetic acid

NADH:

Nicotine adenine dinucletide reduced

MS:

Murashige and Skoog 1962

PAGE:

Polyacryamide gel electrophoresis

PMSF:

Phenylmethylsulphonilfluoride

p-HMB:

p-Hydroxy mercury benzoic acid

PVP:

Polyvinylpyrrolidone

ROS:

Reactive oxygen species

SOD:

Superoxide dismutase

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Authors and Affiliations

  1. Biotechnology Laboratory, Instituto de Desarrollo Regional, UCLM, Campus Universitario s/n, Albacete, 02071, Spain

    Silvia Blazquez & José Antonio Fernández

  2. Department Plant Nutrition, CEBAS (CSIC), P.O. Box 164, 30100, Espinardo, Murcia, Spain

    Enrique Olmos & Nieves Fernández-García

  3. Department Plant Breeding, CEBAS (CSIC), P.O. Box 164, 30100, Espinardo, Murcia, Spain

    José Antonio Hernández & Abel Piqueras

Authors
  1. Silvia Blazquez
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  2. Enrique Olmos
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  3. José Antonio Hernández
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  4. Nieves Fernández-García
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  5. José Antonio Fernández
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  6. Abel Piqueras
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Correspondence to Abel Piqueras.

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Blazquez, S., Olmos, E., Hernández, J.A. et al. Somatic embryogenesis in saffron (Crocus sativus L.). Histological differentiation and implication of some components of the antioxidant enzymatic system. Plant Cell Tiss Organ Cult 97, 49–57 (2009). https://doi.org/10.1007/s11240-009-9497-y

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