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Developmental and hormonal regulation of direct shoot organogenesis and somatic embryogenesis in sugarcane (Saccharum spp. interspecific hybrids) leaf culture

  • Cell Biology and Morphogenesis
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Abstract

Rapid and efficient in vitro regeneration methods that minimise somaclonal variation are critical for the genetic transformation and mass propagation of commercial varieties. Using a transverse thin cell layer culture system, we have identified some of the developmental and physiological constraints that limit high-frequency regeneration in sugarcane leaf tissue. Tissue polarity and consequently the orientation of the explant in culture, size and developmental phase of explant, and auxin concentration play a significant role in determining the organogenic potential of leaf tissue in culture. Both adventitious shoot production and somatic embryogenesis occurred on the proximal cut surface of the explant, and a regeneration gradient, decreasing gradually from the basal to the distal end, exists in the leaf roll. Importantly, auxin, when added to the culture medium, reduced this spatial developmental constraint, as well as the effect of genotype on plant regeneration. Transverse sections (1–2 mm thick) obtained from young leaf spindle rolls and orienting explants with its distal end facing the medium (directly in contact with medium) are critical for maximum regeneration. Shoot regeneration was observed as early as 3 weeks on MS medium supplemented with α-naphthalenencetic acid (NAA) and 6-benzyladenine, while somatic embryogenesis or both adventitious shoot organogenesis and somatic embryogenesis occurred on medium with NAA and chlorophenoxyacetic acid. Twenty shoots or more could be generated from a single transverse section explant. These shoots regenerated roots and successfully established after transplanted to pots. Large numbers of plantlets can be regenerated directly and rapidly using this system. SmartSett®, the registered name for this process and the plants produced, will have significant practical applications for the mass propagation of new cultivars and in genetic modification programs. The SmartSett® system has already been used commercially to produce substantial numbers of plants of orange rust-resistant and new cultivars in Australia.

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Acknowledgements

This Project was funded by Sugar Research and Development Corporation, Australia. The authors thank all the previous and current BSES Limited staff, particularly Michael McKeon, Suzelle Waggett, Kerry Nutt and Zara Borg, for their support and technical assistance. We are grateful to Rhylee Swain, Jeff Smith and Ann Rizzo for providing plant material for this research.

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Author notes

  1. R. Jason Geijskes

    Present address: Queensland University of Technology, School of Life Sciences, 2 George Street, Brisbane, Queensland, 4001, Australia

Authors and Affiliations

  1. David North Plant Research Centre, BSES Limited, 50 Meiers Road, Indooroopilly, Brisbane, Queensland, 4068, Australia

    Prakash Lakshmanan, R. Jason Geijskes, Lifang Wang & Grant R. Smith

  2. CSIRO-Plant Industry, Queensland Bioscience Precinct, 306 Carmody Road, St Lucia, Queensland, 4067, Australia

    Adrian Elliott & Christopher P. L. Grof

  3. Meringa Sugar Experiment Station, BSES Limited, Bruce Highway, Gordonvale, Queensland, 4865, Australia

    Nils Berding

  4. New Zealand Institute for Crop & Food Research Limited, Private Bag 4704, Christchurch, New Zealand

    Grant R. Smith

Authors
  1. Prakash Lakshmanan
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  2. R. Jason Geijskes
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  3. Lifang Wang
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  4. Adrian Elliott
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  5. Christopher P. L. Grof
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  6. Nils Berding
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  7. Grant R. Smith
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Correspondence to Prakash Lakshmanan.

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Communicated by P. P. Kumar

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Lakshmanan, P., Geijskes, R.J., Wang, L. et al. Developmental and hormonal regulation of direct shoot organogenesis and somatic embryogenesis in sugarcane (Saccharum spp. interspecific hybrids) leaf culture. Plant Cell Rep 25, 1007–1015 (2006). https://doi.org/10.1007/s00299-006-0154-1

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  • DOI: https://doi.org/10.1007/s00299-006-0154-1

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