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Development of transgenic pearl millet (Pennisetum glaucum (L.) R. Br.) plants resistant to downy mildew

  • Genetic Transformation and Hybridization
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Abstract

Transgenic pearl millet lines expressing pin gene—exhibiting high resistance to downy mildew pathogen, Sclerospora graminicola—were produced using particle-inflow-gun (PIG) method. Shoot-tip-derived embryogenic calli were co-bombarded with plasmids containing pin and bar genes driven by CaMV 35S promoter. Bombarded calli were cultured on MS medium with phosphinothricin as a selection agent. Primary transformants 1T0, 2T0, and 3T0 showed the presence of both bar and pin coding sequences as evidenced by PCR and Southern blot analysis, respectively. T1 progenies of three primary transformants, when evaluated for downy mildew resistance, segregated into resistant and susceptible phenotypes. T1 plants resistant to downy mildew invariably exhibited tolerance to Basta suggesting co-segregation of pin and bar genes. Further, the downy mildew resistant 1T1 plants were found positive for pin gene in Southern and Northern analyses thereby confirming stable integration, expression, and transmission of pin gene. 1T2 progenies of 1T0 conformed to dihybrid segregation of 15 resistant:1 susceptible plants.

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Abbreviations

2,4-D:

2,4-dichlorophenoxyacetic acid

BAP:

N6-benzylaminopurine

pin :

synthetic prawn antifungal protein encoding gene

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Acknowledgements

We gratefully acknowledge the generous financial support from the Department of Biotechnology, Government of India (New Delhi). We extend our thanks to Dr. V. Panduranga Rao and Dr. R. P. Thakur of ICRISAT for their technical guidance and support for fungal bioassay of pearl millet plants. Research fellowship awarded to AML by the Council of Scientific and Industrial Research, Government of India (New Delhi), is gratefully acknowledged. We thank Mr. M. Suresh Reddy, technical officer, Centre for Plant Molecular Biology, Osmania University, for his unstinted help and co-operation.

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Authors and Affiliations

  1. Centre for Plant Molecular Biology, Osmania University, Hyderabad, 500007, Andhra Pradesh, India

    A. Madhavi Latha, K. V. Rao, T. P. Reddy & V. D. Reddy

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  1. A. Madhavi Latha
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  2. K. V. Rao
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  3. T. P. Reddy
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  4. V. D. Reddy
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Correspondence to V. D. Reddy.

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Communicated by J. C. Register

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Latha, A.M., Rao, K.V., Reddy, T.P. et al. Development of transgenic pearl millet (Pennisetum glaucum (L.) R. Br.) plants resistant to downy mildew. Plant Cell Rep 25, 927–935 (2006). https://doi.org/10.1007/s00299-006-0141-6

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  • DOI: https://doi.org/10.1007/s00299-006-0141-6

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