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Pearl millet transformation system using the positive selectable marker gene phosphomannose isomerase

  • Genetic Transformation and Hybridization
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Abstract

Fertile transgenic pearl millet plants expressing a phosphomannose isomerase (PMI) transgene under control of the maize ubiquitin constitutive promoter were obtained using the transformation system described here. Proliferating immature zygotic embryos were used as target tissue for bombardment using a particle inflow gun. Different culture and selection strategies were assessed in order to obtain an optimised mannose selection protocol. Stable integration of the manA gene into the genome of pearl millet was confirmed by PCR and Southern blot analysis. Stable integration of the manA transgene into the genome of pearl millet was demonstrated in T1 and T2 progeny of two independent transformation events with no more than four to ten copies of the transgene. Similar to results obtained from previous studies with maize and wheat, the manA gene was shown to be a superior selectable marker gene for improving transformation efficiencies when compared to antibiotic or herbicide selectable marker genes.

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Fig. 1
Fig. 2A, B

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Abbreviations

2,4-D::

2,4-Diclorophenoxyacetic acid

IAA::

Indole acetic acid

ICRISAT::

International Crops Research Institute for the Semi-Arid Tropics

IZEs :

Immature zygotic embryos

References

  • Dellaporta SL, Wood J, Hicks JB (1983) A plant DNA minipreparation: version II. Plant Mol Biol Rep 1:19–21

    CAS  Google Scholar 

  • Girgi M, O’Kennedy MM, Morgenstern A, Smith G, Lörz H, Oldach KH (2002) Transgenic and herbicide resistant pearl millet (Pennisetum glaucum L.) R.Br. via microprojectile bombardment of scutellar tissue. Mol Breed 10:243–252

    Article  CAS  Google Scholar 

  • Gless C, Lorz H, Jahne-Gartner A (1998) Establishment of a highly efficient regeneration system from leaf base segments of oat (Avena sativa L.). Plant Cell Rep 17:441–445

    CAS  Google Scholar 

  • Goldman JJ, Hanna WW, Fleming G, Ozias-Akins P (2003) Fertile transgenic pearl millet [Pennisetum glaucum (L.) R. Br.] plants recovered through microprojectile bombardment and phosphinothricin selection of apical meristem-, inflorescence-, and immature embryo-derived embryogenic tissues. Plant Cell Rep 21:999–1009

    Article  CAS  PubMed  Google Scholar 

  • Goldsworthy A, Steet HE (1965) The carbohydrate nutrition of tomato roots. VIII The mechanism of the inhibition by d-mannose of the respiration of excised roots. Ann Bot 29:45–58

    CAS  Google Scholar 

  • Jähne A, Lazzeri PA, Jager-Gussen M, Lorz H (1991) Plant regeneration from embryogenic suspensions derived from anther cultures of barley (Hordeum vulgare L.). Theor Appl Genet 82:74–80

    Google Scholar 

  • Joersbo M, Okkels FT (1996) A novel principle for selection of transgenic plant cells: positive selection. Plant Cell Rep 16:219–221

    Google Scholar 

  • Joersbo M, Donaldson I, Kreiberg J, Petersen SG, Brunstedt J, Okkels FT (1998) Analysis of mannose selection used for transformation of sugar beet. Mol Breed 4:111–117

    Article  CAS  Google Scholar 

  • Joersbo M, Petersen SG, Okkels FT (1999) Parameters interacting with mannose selection employed for the production of transgenic sugar beet. Physiol Plant 105:109–115

    CAS  Google Scholar 

  • Lambé P, Dinant M, Matagne RF (1995) Differential long-term expression and methylation of the hygromycin phosphotransferase (hph) and β-glucuronidase (GUS) genes in transgenic pearl millet (Pennisetum americanum) callus. Plant Sci 108:51–62

    Article  CAS  Google Scholar 

  • Lambé P, Mutambel HSN, Deltour R, Dinant M (1999) Somatic embryogenesis in pearl millet (Pennisetum americanum): strategies to reduce genotype limitation and to maintain long-term totipotency. Plant Cell Tissue Organ Cult 55:23–29

    Article  Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497

    CAS  Google Scholar 

  • Negrotto D, Jolley M, Beer S, Wenck AR, Hansen G (2000) The use of phosphomannose-isomerase as a selectable marker to recover transgenic maize plants (Zea mays L.) via Agrobacterium transformation. Plant Cell Rep 19:798–803

    Google Scholar 

  • O’Kennedy MM, Burger JT, Watson TG (1998) Stable maize transformation using the Particle Inflow Gun. S Afr J Sci 94:188–192

    CAS  Google Scholar 

  • Pego VP, Weisbeek PJ, Smeekens SCM (1999) Mannose inhibits Arabidopsis germination via a hexokinase-mediated step. Plant Physiol 119:1017–1023

    Article  CAS  PubMed  Google Scholar 

  • Pinard F, Chandrapalaiah S (1991) Regeneration of pearl millet explants ICRISAT Cereals Program Annual Report, vol 2, pp 88–89

  • Reed J, Privalle L, Powell ML, Meghji M, Dawson J, Dunder E, Suttie J, Wenck A, Launis K, Kramer C, Chang Y-F, Hansen G, Wright M (2001) Phosphomannose isomerase: an efficient selectable marker for plant transformation. In vitro Cell Dev Biol-Plant 37:127–132

  • Vain P, McMullen MD, Finer J (1993) Osmotic treatment enhances particle bombardment-mediated transient and stable transformation of maize. Plant Cell Rep 12:84–88

    Google Scholar 

  • Wang AS, Evans RA, Altendorf PR, Hanten JA, Doyle MC, Rosichan JL (2000) A mannose selection system for production of fertile transgenic maize plants from protoplasts. Plant Cell Rep 19:654–660

    CAS  Google Scholar 

  • Wright M, Dawson J, Dunder E, Suttie J, Reed J, Kramer C, Chang Y, Novitzky R, Wang H, Artin-Moore L (2001) Efficient biolistic transformation of maize (Zea mays L.) and wheat (Triticum aestivum L.) using the phosphomannose isomerase gene, pmi, as the selectable marker. Plant Cell Rep 20:429–436

    Google Scholar 

  • Zhang P, Puonti-Kaerlas J (2000) PIG-mediated cassava transformation using positive and negative selection. Plant Cell Rep 19:1041–1048

    Google Scholar 

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Acknowledgements

We acknowledge ICRISAT (Zimbabwe) for providing the pearl millet seed material used in this study. We are indebted to Syngenta Crop Protection AG for providing the manA gene construct (Positech system) for this study. We acknowledge the excellent technical assistance of Guguletho Ngwenya, Rufus Khonothi, Jeffrey Mathabe and Moses Mokoena.

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Authors and Affiliations

  1. CSIR Food, Biological and Chemical Technologies (Bio/Chemtek), P.O. Box 395, 0001, Pretoria, South Africa

    M.M. O’Kennedy

  2. Institute for Plant Biotechnology, Stellenbosch University, Private Bag X1, 7602, Matieland, South Africa

    M.M. O’Kennedy & F. C. Botha

  3. Department of Genetics, Stellenbosch University, Private Bag X1, 7602, Matieland, South Africa

    J. T. Burger

  4. South African Sugar Association Experiment Station, Private Bag X02, Mount Edgecombe, South Africa

    F. C. Botha

Authors
  1. M.M. O’Kennedy
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  2. J. T. Burger
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  3. F. C. Botha
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Correspondence to M.M. O’Kennedy.

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Communicated by H. Lörz

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O’Kennedy, M., Burger, J.T. & Botha, F.C. Pearl millet transformation system using the positive selectable marker gene phosphomannose isomerase. Plant Cell Rep 22, 684–690 (2004). https://doi.org/10.1007/s00299-003-0746-y

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  • DOI: https://doi.org/10.1007/s00299-003-0746-y

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