Current Microbiology

, Volume 49, Issue 5, pp 372–375

Indirect Immunofluorescence Microscopy for Direct Detection of Xylella fastidiosa in Xylem Sap

  • Dulce Carbajal
  • Kevin A. Morano
  • Lisa D. Morano
Article

Abstract

The plant pathogen Xylella fastidiosa is the causative agent of a number of diseases of economically important crops, including Pierce’s disease that affects grapevines. Using a commercially available antibody specific for X. fastidiosa, we have established a protocol for microscopic identification of the bacterium by indirect immunofluorescence. This antibody clearly labels an uncharacterized antigen concentrated at a single pole of X. fastidiosa cells, but does not react with a non-Xylella control. This technique was also performed successfully on xylem exudates from several different plant genera and correlated well with standard enzyme-linked immunosorbent assay tests. These results establish a novel method for in situ assessment of X. fastidiosa infection from host plants.

Literature Cited

  1. 1.
    Banks, D, Albibi, R, Chen, J, Lamikanra, O, Jarret, RL, Smith, BJ 1999Specific detection of Xylella fastidiosa Pierce’s disease strainsCurr Microbiol398588CrossRefPubMedGoogle Scholar
  2. 2.
    Chen, J, Lamikanra, O, Chang, CJ, Hopkins, DL 1995Randomly amplified polymorphic DNA analysis of Xylella fastidiosa Pierce’s disease and oak leaf scorch pathotypesAppl Environ Microbiol6116881690PubMedGoogle Scholar
  3. 3.
    Coletta-Filho, HD, Machado, MA 2002Evaluation of the genetic structure of Xylella fastidiosa populations from different Citrus sinensis varietiesAppl Environ Microbiol6837313736CrossRefPubMedGoogle Scholar
  4. 4.
    da Costa, PI, Franco, CF, Miranda, VS, Teixeira, DC, Hartung, JS 2000Strains of Xylella fastidiosa rapidly distinguished by arbitrarily primed-PCRCurr Microbiol40279282CrossRefPubMedGoogle Scholar
  5. 5.
    Davis, MJ, French, WJ, Shaad, NW 1981Axenic culture of the bacteria associated with phony disease of peach and plum leaf scaldCurr Microbiol6309314Google Scholar
  6. 6.
    French, WJ, Stassi, DL, Schaad, NW 1978The use of immunofluorescence for the identification of phony peach bacteriumPhytopathol6811061108Google Scholar
  7. 7.
    Hendson, M, Purcell, AH, Chen, D, Smart, C, Guilhabert, M, Kirkpatrick, B 2001Genetic diversity of Pierce’s disease strains and other pathotypes of Xylella fastidiosaApp Environ Microbiol67895903CrossRefGoogle Scholar
  8. 8.
    Hewitt, WB 1958The probable home of Pierce’s disease virusPlant Dis Rep42211215Google Scholar
  9. 9.
    Hill, BL, Purcell, AH 1995Multiplication and movement of Xylella fastidiosa within grapevine and four other plantsPhytopathol8513681372Google Scholar
  10. 10.
    Hill, BL, Purcell, AH 1997Populations of Xylella fastidiosa in plants required for transmission by an efficient vectorPhytopathol8711971201Google Scholar
  11. 11.
    Hopkins, DL, Bistline, FW, Russo, LW, Thompson, CM 1991Seasonal fluctuation in the occurrence of Xylella fastidiosa in root and stem extracts from citrus with blightPlant Dis75145147Google Scholar
  12. 12.
    Hopkins, DL, Purcell, AH 2002Xylella fastidiosa: cause of Pierce’s disease of grapevine and other emergent diseasesPlant Dis8610561066Google Scholar
  13. 13.
    Minsavage, GV, Thompson, CM, Hopkins, DL, Leite, RMVBC, Stall, RE 1994Development of a polymerase chain reaction protocol for detection of Xylella fastidiosa in plant tissuePhytopathol84456461Google Scholar
  14. 14.
    Pooler, MR, Hartung, JS 1995Genetic relationships among strains of Xylella fastidiosa from RAPD-PCR dataCurr Microbiol31134137CrossRefPubMedGoogle Scholar
  15. 15.
    Pooler, MR, Myung, IS, Bentz, J, Sherald, J, Hartung, JS 1997Detection of Xylella fastidiosa in potential insect vectors by immunomagnetic separation and nested polymerase chain reactionLett Appl Microbiol25123126CrossRefPubMedGoogle Scholar
  16. 16.
    Schaad, NW, Opgenorth, D, Gaush, P 2002Real-time polymerase chain reaction for one-hour on-site diagnosis of Pierce’s disease of grape in early season asymptomatic vinesPhytopathol92721728Google Scholar
  17. 17.
    Yu, XC, Margolin, W 1999FtsZ ring clusters in min and partition mutants: role of both the Min system and the nucleoid in regulating FtsZ ring localizationMol Microbiol32315326CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, Inc. 2004

Authors and Affiliations

  • Dulce Carbajal
    • 1
  • Kevin A. Morano
    • 2
  • Lisa D. Morano
    • 1
  1. 1.Department of Natural SciencesUniversity of Houston-DowntownHoustonUSA
  2. 2.Department of Microbiology and Molecular GeneticsUniversity of Texas Medical SchoolHoustonUSA

Personalised recommendations