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Enhanced resistance to a lepidopteran pest in transgenic sugar beet plants expressing synthetic cry1Ab gene

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Abstract

Two diploid sugar beet genotypes of agronomical importance were transformed using Agrobactrium tumefaciens harboring pBI35Scry containing a synthetic cry1Ab gene. Leaf blade with attached shoot bases, a highly regenerative tissue, were used as explant substratum for transformation. PCR screening with cry1Ab-specific primers showed the presence of transgene in more than 50% of the regenerated kanamycin-resistant plants after treatment with the antibiotic. A transformation rate of 8.8–12.2% (depending on genotype) was achieved as revealed by genomic DNA dot blotting. The intact integration of transgene cassette into the genome was furthermore confirmed by Southern blot analysis. The expression of the cry1Ab gene encoding a truncated endotoxin (67 kDa) at about 0.1% of total soluble protein was achieved in the leaves of transgenic plants as shown by Western blot analysis. Bioassays under in vitro conditions with Spodoptera littoralis, one of the most important pests in sugar beet fields, demonstrated enhanced resistance against this pest. The inheritance of the inserted transgene was confirmed in F1 plants obtained through crossing of T0 plants with a cytoplasmic male sterile line. Transgenic plants are currently grown in a greenhouse and will be subjected to further bioassay analyses against other lepidopteran pests of sugar beet.

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Acknowledgment

This research was jointly funded by Agricultural Biotechnology Research Institute of Iran (ABRII) and Sugar Beet Seed Institute of Iran (SBSI). We wish to appreciate Dr. AA Habashi (ABRII) for his great supports. We would like to thank Dr. SE Mortazavi (ABRII) for his technical advice as well. Authors wish to acknowledge Dr. N Amman, Technical University of Delft, Netherlands and Dr. A Rajabi, Sugar Beet Seed Institute of Iran, for the critical review and their suggestions for the improvement of the manuscript.

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Authors and Affiliations

  1. Department of Agronomy and Plant Breeding, University of Urmia, Urmia, Iran

    Morad Jafari

  2. Department of Agronomy and Plant Breeding, University of Tabriz, Tabriz, Iran

    Morad Jafari, Mostafa Valizadeh & Seyed Aboulghasem Mohammadi

  3. Sugar Beet Seed Institute (SBSI), P.O. Box 31585-4114, Karaj, Iran

    Peyman Norouzi

  4. National Institute of Genetic Engineering and Biotechnology (NIGEB), P.O. Box 14155-6343, Tehran, Iran

    Mohammad Ali Malboobi

  5. Agricultural Biotechnology Research Institute of Iran (ABRII), Karaj, Iran

    Behzad Ghareyazie

  6. Plant Protection Research Institute of Iran, Tehran, Iran

    Mozhgan Mousavi

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  1. Morad Jafari
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Correspondence to Peyman Norouzi.

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Jafari, M., Norouzi, P., Malboobi, M.A. et al. Enhanced resistance to a lepidopteran pest in transgenic sugar beet plants expressing synthetic cry1Ab gene. Euphytica 165, 333–344 (2009). https://doi.org/10.1007/s10681-008-9792-4

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