Infection of the Circulifer haematoceps cell line Ciha-1 by Spiroplasma citri: the non-insect-transmissible strain 44 is impaired in invasion.
Successful transmission of Spiroplasma citri by its leafhopper vector requires a specific interaction between the spiroplasma surface and the insect cells. With the aim of studying these interactions at the cellular and molecular levels, a cell line, named Ciha-1, was established using embryonic tissues from the eggs of the S. citri natural vector Circulifer haematoceps. This is the first report, to our knowledge, of a cell line for this leafhopper species and of its successful infection by the insect-transmissible strain S. citri GII3. Adherence of the spiroplasmas to the cultured Ciha-1 cells was studied by c.f.u. counts and by electron microscopy. Entry of the spiroplasmas into the insect cells was analysed quantitatively by gentamicin protection assays and qualitatively by double immunofluorescence microscopy. Spiroplasmas were detected within the cell cytoplasm as early as 1 h after inoculation and survived at least 2 days inside the cells. Comparing the insect-transmissible GII3 and non-insect-transmissible 44 strains revealed that adherence to and entry into Ciha-1 cells of S. citri 44 were significantly less efficient than those of S. citri GII3.
Duret
Sybille
S
Université de Bordeaux 2, UMR 1090 Génomique Diversité et Pouvoir Pathogène, F-33883 Villenave d'Ornon, France.|INRA, Centre de Bordeaux-Aquitaine, UMR 1090 Génomique Diversité et Pouvoir Pathogène, F-33883 Villenave d'Ornon, France.
Batailler
Brigitte
B
Plateau Technique Imagerie/Cytologie, INRA, Centre de Bordeaux-Aquitaine, F-33883 Villenave d'Ornon, France.|Université de Bordeaux 2, UMR 1090 Génomique Diversité et Pouvoir Pathogène, F-33883 Villenave d'Ornon, France.|INRA, Centre de Bordeaux-Aquitaine, UMR 1090 Génomique Diversité et Pouvoir Pathogène, F-33883 Villenave d'Ornon, France.
Danet
Jean-Luc
JL
Université de Bordeaux 2, UMR 1090 Génomique Diversité et Pouvoir Pathogène, F-33883 Villenave d'Ornon, France.|INRA, Centre de Bordeaux-Aquitaine, UMR 1090 Génomique Diversité et Pouvoir Pathogène, F-33883 Villenave d'Ornon, France.
Béven
Laure
L
Université de Bordeaux 2, UMR 1090 Génomique Diversité et Pouvoir Pathogène, F-33883 Villenave d'Ornon, France.|INRA, Centre de Bordeaux-Aquitaine, UMR 1090 Génomique Diversité et Pouvoir Pathogène, F-33883 Villenave d'Ornon, France.
Renaudin
Joël
J
Université de Bordeaux 2, UMR 1090 Génomique Diversité et Pouvoir Pathogène, F-33883 Villenave d'Ornon, France.|INRA, Centre de Bordeaux-Aquitaine, UMR 1090 Génomique Diversité et Pouvoir Pathogène, F-33883 Villenave d'Ornon, France.
Arricau-Bouvery
Nathalie
N
Université de Bordeaux 2, UMR 1090 Génomique Diversité et Pouvoir Pathogène, F-33883 Villenave d'Ornon, France.|INRA, Centre de Bordeaux-Aquitaine, UMR 1090 Génomique Diversité et Pouvoir Pathogène, F-33883 Villenave d'Ornon, France.
D000818:Animals; D002460:Cell Line / Q000382:microbiology*; D006430:Hemiptera / Q000382:microbiology*; D007303:Insect Vectors / Q000382:microbiology*; D010935:Plant Diseases / Q000382:microbiology; D045642:Spiroplasma citri / Q000472:pathogenicity* / Q000502:physiology; D014774:Virulence