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Entry of Spiroplasma citri into Circulifer haematoceps cells involves interaction between spiroplasma

Fabien Labroussaa1, Nathalie Arricau-Bouvery, Marie-Pierre Dubrana

  • 1UMR 1090 Génomique Diversité Pouvoir Pathogène, INRA, Université Victor Ségalen Bordeaux 2, 71 Avenue Edouard Bourlaux BP 81, F-33883 Villenave d'Ornon, France.

Applied and Environmental Microbiology
|February 2, 2010
PubMed

Insights

Spiroplasma citri uses phosphoglycerate kinase (PGK) to enter insect cells. This study shows that PGK inhibits spiroplasma internalization, a key step in plant disease transmission by insect vectors.

Area of Science:

  • Plant pathology
  • Insect vector biology
  • Molecular microbiology

Background:

  • Phytopathogenic mollicutes, including spiroplasmas, rely on insect vectors for transmission.
  • Successful transmission involves mollicutes penetrating insect gut, multiplying, and traversing salivary glands.
  • Molecular interactions between mollicutes and vectors are crucial for regulating transmission.

Purpose of the Study:

  • To investigate molecular interactions between Spiroplasma citri and its leafhopper vector, Circulifer haematoceps.
  • To identify specific proteins involved in the interaction and their role in transmission.
  • To elucidate the mechanism by which spiroplasmas enter insect cells.

Main Methods:

  • In vitro protein overlay assays to identify protein binding.
  • Liquid chromatography-tandem mass spectrometry (LC-MS/MS) for protein identification.
  • Confocal microscopy to visualize spiroplasma and host protein colocalization.
  • Competitive binding and internalization assays using cultured leafhopper cells.

Main Results:

  • Identified binding activities between Spiroplasma citri proteins and salivary gland proteins, including actin.
  • Isolated and identified Spiroplasma citri phosphoglycerate kinase (PGK) as an actin-binding protein.
  • Observed colocalization of spiroplasmas with actin filaments in infected salivary glands.
  • Demonstrated that exogenous PGK inhibits Spiroplasma citri internalization into leafhopper cells in a dose-dependent manner, without affecting attachment.

Conclusions:

  • Phosphoglycerate kinase (PGK) plays a critical role in the internalization of Spiroplasma citri into its insect vector.
  • The interaction between Spiroplasma citri PGK and host actin is essential for spiroplasma entry into eukaryotic cells.
  • Understanding this interaction can lead to novel strategies for controlling plant diseases transmitted by insect vectors.

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