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Role of vitronectin-like protein in Agrobacterium attachment and transformation of Arabidopsis cells

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Abstract

The role of plant vitronectin-like protein (Vn) in Agrobacterium–host plant interactions and receptor-specific bacterial attachment is unclear and still open to debate. Using a well-established Agrobacterium-mediated Arabidopsis transformation system, the marker gene β-glucuronidase (GUS) of Escherichia coli, and biochemical and cytological methods, such as ELISA tests, immunoblots, immunolocalization, and functional in vitro binding assays, we have reassessed the role of Vn in receptor-specific bacterial attachment and transformation. We provide evidence that Vn is present in the host plant cells and anti-human vitronectin antibody cross-reacts with a 65-kDa protein from Arabidopsis cells. The specificity of the immunological cross-reactivity of anti-vitronectin antibodies was further demonstrated by ELISA competition experiments. Immunogold labeling showed that Vn is localized in the plant cell wall, and its level increased considerably after phytohormone treatment of the petiole explants. However, Agrobacterium attachment was unaffected, and no inhibition of petiole cell transformation was detected in the presence of human vitronectin and anti-vitronectin antibodies in the media. Additionally, no correlation between the occurrence of Vn, attachment of bacteria to the cells, and susceptibility to Agrobacterium-mediated transformation was observed. Taken together, our data do not support a functional role of plant Vn as the receptor for site-specific Agrobacterium attachment leading to the transformation of Arabidopsis cells.

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Abbreviations

ELISA:

Enzyme-linked immunosorbent assays

GUS:

β-glucuronidase

Vn:

Plant vitronectin-like protein

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Acknowledgments

We thank Zoulikha Zaraoui and Gérard Vasseur for technical assistance and plant maintenance. We also thank Professor. F. Guerineau and Dr. S. Millam for critically reading the manuscript. This work was supported by the French Ministry for Research and Higher Education (MENSER) and also by the Regional Government of Picardie, France.

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Authors and Affiliations

  1. Unité de Recherche EA3900 “Biologie des Plantes et Contrôle des Insectes ravageurs”, Laboratoire Androgenèse et Biotechnologie, Faculté des Sciences, Université de Picardie Jules Verne, 33 rue Saint-Leu, 80039, Amiens, France

    Hélène Clauce-Coupel, Sophie Chateau, Corinne Ducrocq, Vincent Niot, Frédéric Dubois, Brigitte Sangwan-Norreel & Rajbir S. Sangwan

  2. INSERM–U 872, Centre de Recherche des Cordeliers 15, Rue de l’Ecole de Médecine, 75006, Paris, France

    Srini Kaveri

Authors
  1. Hélène Clauce-Coupel
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  2. Sophie Chateau
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  3. Corinne Ducrocq
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  4. Vincent Niot
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  5. Srini Kaveri
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  6. Frédéric Dubois
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  7. Brigitte Sangwan-Norreel
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  8. Rajbir S. Sangwan
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Correspondence to Rajbir S. Sangwan.

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Clauce-Coupel, H., Chateau, S., Ducrocq, C. et al. Role of vitronectin-like protein in Agrobacterium attachment and transformation of Arabidopsis cells. Protoplasma 234, 65–75 (2008). https://doi.org/10.1007/s00709-008-0022-7

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  • DOI: https://doi.org/10.1007/s00709-008-0022-7

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