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Evaluation of haemoglobin (erythrogen): for improved somatic embryogenesis and plant regeneration in cotton (Gossypium hirsutum L. cv. SVPR 2)

  • Cell Biology and Morphogenesis
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Abstract

Somatic embryogenesis in cotton (Gossypium hirsutum L.) is accelerated when the plant regeneration medium is supplemented with haemoglobin (erythrogen). In cotton SVPR 2 lines, a higher frequency of embryoid formation was observed when the medium contained 400 mg/l haemoglobin. Fresh weight of the callus, rate of embryoid induction, number of embryoids formed and the percentage of plant regeneration from somatic embryos were increased. Among the two different cultivars tested, MCU 11 showed no response to the presence of haemoglobin when compared to SVPR 2, and embryogenic callus formation was completely absent in the former. Medium containing MS salts, 100 mg/l myo-inositol , 0.3 mg/l thiamine-HCL, 0.3 mg/l Picloram (PIC), 0.1 mg/l kinetin and 400 mg/l haemoglobin effected a better response with respect to embryogenic callus induction. After 8 weeks of culture, a high frequency of embryoid induction was observed on medium containing MS basal salts, 100 mg/l myo-inositol, 0.3 mg/l PIC , 0.1 mg/l isopentenyl adenine, 1.0 g/l NH4NO3 and 400 mg/l haemoglobin. Plant regeneration was observed in 75.8% of the mature somatic embryos, and whole plant regeneration was achieved within 6–7 months of culture. The regenerated plantlets were fertile and similar to in vivo-grown, seed-derived plants except that they were phenotypically smaller. A positive influence of haemoglobin was observed at concentrations up to 400 mg/l at all stages of somatic embryogenesis. The increase in the levels of antioxidant enzyme activities, for example superoxide dismutase and peroxidase, indicated the presence of excess oxygen uptake and the stressed condition of the plant tissues that arose from haemoglobin supplementation. This increased oxygen uptake and haemoglobin-mediated stress appeared to accelerate somatic embryogenesis in cotton.

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Fig. 1a–I

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Abbreviations

BAP:

Benzylaminopurine

2,4-D:

2,4-Dichlorophenoxyacetic acid

GA3:

Gibberellic acid

GR:

Glutathione reductase

2iP:

Isopentenyl adenine

KT:

Kinetin

NAA:

Naphthaleneacetic acid

PFC:

Perfluorocarbon

PIC:

Picloram

PO:

Peroxidase

ROS:

Reactive oxygen species

SOD:

Superoxide dismutase

T.HCl:

Thiamine hydrochloride

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Acknowledgements

The authors are grateful to Drs. P. Anthony, M.R. Davey and J.B. Power, Plant Science Division, School of Bio-sciences, Sutton Bonnington Campus, University of Nottingham, Loughborough, UK for their useful suggestions and comments during this research work. We also thank Dr. Wendy Harwood, Department of Crop Genetics, John Innes Centre, Norwich, UK for language correction and improvement of this manuscript.

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  1. Department of Plant Science, Bharathidasan University, Tiruchirappalli, 620 024, Tamil Nadu, India

    M. Ganesan & N. Jayabalan

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  1. M. Ganesan
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  2. N. Jayabalan
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Correspondence to M. Ganesan.

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Communicated by D. Bartels

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Ganesan, M., Jayabalan, N. Evaluation of haemoglobin (erythrogen): for improved somatic embryogenesis and plant regeneration in cotton (Gossypium hirsutum L. cv. SVPR 2). Plant Cell Rep 23, 181–187 (2004). https://doi.org/10.1007/s00299-004-0822-y

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  • DOI: https://doi.org/10.1007/s00299-004-0822-y

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