1887

Abstract

Successful transmission of 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 natural vector . 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 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 44 were significantly less efficient than those of GII3.

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.035063-0
2010-04-01
2024-03-29
Loading full text...

Full text loading...

/deliver/fulltext/micro/156/4/1097.html?itemId=/content/journal/micro/10.1099/mic.0.035063-0&mimeType=html&fmt=ahah

References

  1. Ammar E-D., Hogenhout S. A. 2005; Use of immunofluorescence confocal laser scanning microscopy to study distribution of the bacterium corn stunt spiroplasma in vector leafhoppers (Hemiptera: Cicadellidae) and in host plants. Ann Entomol Soc Am 98:820–826
    [Google Scholar]
  2. Ammar E-D., Fulton D., Bai X., Meulia T., Hogenhout S. A. 2004; An attachment tip and pili-like structures in insect- and plant-pathogenic spiroplasmas of the class Mollicutes. Arch Microbiol 181:97–105
    [Google Scholar]
  3. Amorena B., Baselga R., Aguilar B. 1990; Factors influencing the degree of in vitro bacterial adhesion to ovine mammary gland epithelial cells. Vet Microbiol 24:43–53
    [Google Scholar]
  4. Andreev J., Borovsky Z., Rosenshine I., Rottem S. 1995; Invasion of HeLa cells by Mycoplasma penetrans and the induction of tyrosine phosphorylation of a 145-kDa host cell protein. FEMS Microbiol Lett 132:189–194
    [Google Scholar]
  5. Baseman J. B., Lange M., Criscimagna N. L., Giron J. A., Thomas C. A. 1995; Interplay between mycoplasmas and host target cells. Microb Pathog 19:105–116
    [Google Scholar]
  6. Berg M., Melcher U., Fletcher J. 2001; Characterization of Spiroplasma citri adhesion related protein SARP1, which contains a domain of a novel family designated sarpin. Gene 275:57–64
    [Google Scholar]
  7. Berho N., Duret S., Danet J. L., Renaudin J. 2006a; Plasmid pSci6 from Spiroplasma citri GII-3 confers insect transmissibility to the non-transmissible strain S. citri 44. Microbiology 152:2703–2716
    [Google Scholar]
  8. Berho N., Duret S., Renaudin J. 2006b; Absence of plasmids encoding adhesion-related proteins in non-insect-transmissible strains of Spiroplasma citri. Microbiology 152:873–886
    [Google Scholar]
  9. Boutareaud A., Danet J. L., Garnier M., Saillard C. 2004; Disruption of a gene predicted to encode a solute binding protein of an ABC transporter reduces transmission of Spiroplasma citri by the leafhopper Circulifer haematoceps. Appl Environ Microbiol 70:3960–3967
    [Google Scholar]
  10. Bové J. M. 1997; Spiroplasmas: infections agents of plants, arthropods and vertebrates. Wien Klin Wochenschr 109:604–612
    [Google Scholar]
  11. Bové J. M., Renaudin J., Saillard C., Foissac X., Garnier M. 2003; Spiroplasma citri, a plant pathogenic mollicute: relationships with its two hosts, the plant and the leafhopper vector. Annu Rev Phytopathol 41:483–500
    [Google Scholar]
  12. Boyle E. C., Brown N. F., Brumell J. H., Finlay B. B. 2007; Src homology domain 2 adaptors affect adherence of Salmonella enterica serovar Typhimurium to non-phagocytic cells. Microbiology 153:3517–3526
    [Google Scholar]
  13. Burnett T. A., Dinkla K., Rohde M., Chhatwal G. S., Uphoff C., Srivastava M., Cordwell S. J., Geary S., Liao X. other authors 2006; P159 is a proteolytically processed, surface adhesin of Mycoplasma hyopneumoniae: defined domains of P159 bind heparin and promote adherence to eukaryote cells. Mol Microbiol 60:669–686
    [Google Scholar]
  14. Cherry S. 2008; Genomic RNAi screening in Drosophila S2 cells: what have we learned about host–pathogen interactions?. Curr Opin Microbiol 11:262–270
    [Google Scholar]
  15. Dessi D., Delogu G., Emonte E., Catania M. R., Fiori P. L., Rappelli P. 2005; Long-term survival and intracellular replication of Mycoplasma hominis in Trichomonas vaginalis cells: potential role of the protozoon in transmitting bacterial infection. Infect Immun 73:1180–1186
    [Google Scholar]
  16. Drasbek M., Christiansen G., Drasbek K. R., Holm A., Birkelund S. 2007; Interaction between the P1 protein of Mycoplasma pneumoniae and receptors on HEp-2 cells. Microbiology 153:3791–3799
    [Google Scholar]
  17. Duret S., Berho N., Danet J. L., Garnier M., Renaudin J. 2003; Spiralin is not essential for helicity, motility, or pathogenicity but is required for efficient transmission of Spiroplasma citri by its leafhopper vector Circulifer haematoceps. Appl Environ Microbiol 69:6225–6234
    [Google Scholar]
  18. Felek S., Lawrenz M. B., Krukonis E. S. 2008; The Yersinia pestis autotransporter YapC mediates host cell binding, autoaggregation and biofilm formation. Microbiology 154:1802–1812
    [Google Scholar]
  19. Fletcher J., Schultz G. A., Davis R. E., Eastman C. E., Goodman R. M. 1981; Brittle root disease of horseradish: evidence for an etiological role of Spiroplasma citri. Phytopathology 71:1073–1080
    [Google Scholar]
  20. Fletcher J., Wayadande A., Melcher U., Ye F. 1998; The phytopathogenic mollicute–insect vector interface: a closer look. Phytopathology 88:1351–1358
    [Google Scholar]
  21. Fleury B., Bergonier D., Berthelot X., Peterhans E., Frey J., Vilei E. M. 2002; Characterization of P40, a cytadhesin of Mycoplasma agalactiae. Infect Immun 70:5612–5621
    [Google Scholar]
  22. Foissac X., Danet J. L., Saillard C., Whitcomb R. F., Bové J. M. 1996; Experimental infections of plant by spiroplasmas. In Molecular and Diagnostic Procedures in Mycoplasmology vol. 2 pp 385–389 Edited by Razin S., Tully J. G. New York: Academic Press;
    [Google Scholar]
  23. Fos A., Bové J. M., Lallemand J., Saillard C., Vignault J. C., Ali Y., Brun P., Vogel R. 1986; The leafhopper Neoaliturus haematoceps (Mulsant & Rey) is a vector of Spiroplasma citri in the Mediterranean. Ann Inst Pasteur Microbiol 137A:97–107
    [Google Scholar]
  24. Gaurivaud P., Danet J. L., Laigret F., Garnier M., Bové J. M. 2000; Fructose utilization and phytopathogenicity of Spiroplasma citri. Mol Plant Microbe Interact 13:1145–1155
    [Google Scholar]
  25. Giron J. A., Lange M., Baseman J. B. 1996; Adherence, fibronectin binding, and induction of cytoskeleton reorganization in cultured human cells by Mycoplasma penetrans. Infect Immun 64:197–208
    [Google Scholar]
  26. Heesemann J., Laufs R. 1985; Double immunofluorescence microscopic technique for accurate differentiation of extracellularly and intracellularly located bacteria in cell culture. J Clin Microbiol 22:168–175
    [Google Scholar]
  27. Hosseini Pour A. 2000 Determination of some molecular and cellular characteristics of Spiroplasma citri, the causal agent of citrus stubborn disease in Kerman, Fars and Mazadaran provinces PhD thesis Tarbiat Modares University; Tehran:
    [Google Scholar]
  28. Hunter W. B., Hsu H. T. 1996; Formulation of an insect medium for thrips monolayer cell cultures (Thysanoptera: Thripidae: Frankliniella occidentalis. J Invertebr Pathol 67:125–128
    [Google Scholar]
  29. Joshi B. D., Berg M., Rogers J., Fletcher J., Melcher U. 2005; Sequence comparisons of plasmids pBJS-O of Spiroplasma citri and pSKU146 of S. kunkelii: implications for plasmid evolution. BMC Genomics 6:175
    [Google Scholar]
  30. Killiny N., Castroviejo M., Saillard C. 2005; Spiroplasma citri spiralin acts in vitro as a lectin binding to glycoproteins from its insect vector Circulifer haematoceps. Phytopathology 95:541–548
    [Google Scholar]
  31. Killiny N., Batailler B., Foissac X., Saillard C. 2006; Identification of a Spiroplasma citri hydrophilic protein associated with insect transmissibility. Microbiology 152:1221–1230
    [Google Scholar]
  32. Kwon M. O., Wayadande A. C., Fletcher J. 1999; Spiroplasma citri movement into the intestines and salivary glands of its leafhopper vector, Circulifer tenellus. Phytopathology 89:1144–1151
    [Google Scholar]
  33. Liu H. Y., Gumpf D. J., Oldfield G. N., Calavan E. C. 1983; Transmission of Spiroplasma citri by Circulifer tenellus. Phytopathology 73:582–585
    [Google Scholar]
  34. Maixner M., Ahrens U., Seemüller E. 1995; Detection of the German grapevine yellows (Vergilbungskrankheit) MLO in grapevine, alternative hosts and a vector by a specific PCR procedure. Eur J Plant Pathol 101:241–250
    [Google Scholar]
  35. Maramorosch K. 1979; Leafhopper tissue culture. In Leafhopper Vectors and Plant Disease Agents pp 484–511 Edited by Maramorosch K., Harris K. F. New York: Academic Press;
    [Google Scholar]
  36. Martínez-Salgado C., López-Hernández F. J., López-Novoa J. M. 2007; Glomerular nephrotoxicity of aminoglycosides. Toxicol Appl Pharmacol 223:86–98
    [Google Scholar]
  37. Omura T., Kimura I. 1994; Leafhopper cell culture for virus research. In Arthropod Cell Culture Systems pp 91–107 Edited by Maramorosch K., McIntosh A. H. Boca Raton: CRC Press;
    [Google Scholar]
  38. Ozbek E., Miller S. A., Meulia T., Hogenhout S. A. 2003; Infection and replication sites of Spiroplasma kunkelii (Class: Mollicutes) in midgut and Malpighian tubules of the leafhopper Dalbulus maidis. J Invertebr Pathol 82:167–175
    [Google Scholar]
  39. Renaudin J., Lartigue C. 2005; OriC plasmids as gene vectors for mollicutes. In Mycoplasmas: Pathogenesis, Molecular Biology, and Emerging Strategies for Control pp 3–30 Edited by Blanchard A., Browning G. Norwich, UK: Horizon Scientific Press;
    [Google Scholar]
  40. Rottem S. 2003; Interaction of Mycoplasmas with host cells. Physiol Rev 83:417–432
    [Google Scholar]
  41. Saglio P., Laflèche D., Bonissol C., Bové J. M. 1971; Isolement et culture in vitro des mycoplasmes associés au stubborn des agrumes et leur observation au microscope électronique. C R Acad Sci Hebd Seances Acad Sci D 272:1387–1390 in French
    [Google Scholar]
  42. Saglio P., Lhospital M., Laflèche D., Dupont G., Bové J. M., Tully J. G., Freundt E. A. 1973; Spiroplasma citri gen. and sp. n.: a mycoplasma-like organism associated with “stubborn” disease of citrus. Int J Syst Bacteriol 23:191–204
    [Google Scholar]
  43. Saillard C., Carle P., Duret-Nurbel S., Henri R., Killiny N., Carrère S., Gouzy J., Bové J. M., Renaudin J., Foissac X. 2008; The abundant extrachromosomal DNA content of the Spiroplasma citri GII3–3X genome. BMC Genomics 9:195
    [Google Scholar]
  44. Steiner T., McGarrity G. J., Bové J. M., Phillips D. M., Garnier M. 1984; Insect cell cultures in the study of attachment and pathogenicity of spiroplasmas and mycoplasmas. Ann Microbiol (Paris ) 135A:47–53
    [Google Scholar]
  45. Svenstrup H. F., Nielsen P. K., Drasbek M., Birkelund S., Christiansen G. 2002; Adhesion and inhibition assay of Mycoplasma genitalium and M. pneumoniae by immunofluorescence microscopy. J Med Microbiol 51:361–373
    [Google Scholar]
  46. Takiya D. M., Tran P. L., Dietrich C. H., Moran N. A. 2006; Co-cladogenesis spanning three phyla: leafhoppers (Insecta: Hemiptera: Cicadellidae) and their dual bacterial symbionts. Mol Ecol 15:4175–4191
    [Google Scholar]
  47. Veiga E., Cossart P. 2006; The role of clathrin-dependent endocytosis in bacterial internalization. Trends Cell Biol 16:499–504
    [Google Scholar]
  48. Vignault J. C., Bové J. M., Saillard C., Vogel R., Faro A., Venegas A., Stemmer W., Aoki S., McRoy R. E. other authors 1980; Mise en culture de spiroplasmes à partir de matériel végétal et d'insectes provenant de pays circum méditerranéens et du Proche Orient. C R Acad Sci Hebd Seances Acad Sci D 290:775–780 in French
    [Google Scholar]
  49. Wayadande A. C., Fletcher J. 1998; Development and use of an established cell line of the leafhopper Circulifer tenellus to characterize Spiroplasma citri–vector interactions. J Invertebr Pathol 72:126–131
    [Google Scholar]
  50. Whitcomb R. F. 1983; Culture media for spiroplasma. In Methods in Mycoplasmology vol. I pp 147–158 Edited by Razin S., Tully J. G. New York: Academic Press;
    [Google Scholar]
  51. Winner F., Rosengarten R., Citti C. 2000; In vitro cell invasion of Mycoplasma gallisepticum. Infect Immun 68:4238–4244
    [Google Scholar]
  52. Yavlovich A., Tarshis M., Rottem S. 2004; Internalization and intracellular survival of Mycoplasma pneumoniae by non-phagocytic cells. FEMS Microbiol Lett 233:241–246
    [Google Scholar]
  53. Yu J., Wayadande A. C., Fletcher J. 2000; Spiroplasma citri surface protein P89 implicated in adhesion to cells of the vector Circulifer tenellus. Phytopathology 90:716–722
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.035063-0
Loading
/content/journal/micro/10.1099/mic.0.035063-0
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error