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Journal of Bacteriology, August 2004, p. 5442-5449, Vol. 186, No. 16
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.16.5442-5449.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

DNA Microarray-Based Genome Comparison of a Pathogenic and a Nonpathogenic Strain of Xylella fastidiosa Delineates Genes Important for Bacterial Virulence{dagger}

Tie Koide,1,{dagger} Paulo A. Zaini,1,{ddagger} Leandro M. Moreira,1 Ricardo Z. N. Vêncio,2 Adriana Y. Matsukuma,1 Alan M. Durham,3 Diva C. Teixeira,4 Hamza El-Dorry,1 Patrícia B. Monteiro,4,§ Ana Claudia R. da Silva,1 Sergio Verjovski-Almeida,1 Aline M. da Silva,1* and Suely L. Gomes1*

Departamento de Bioquímica, Instituto de Química,1 Departamento de Estatística,2 Ciência da Computação, Instituto de Matemática e Estatística, Universidade de São Paulo, São Paulo,3 Fundo de Defesa da Citricultura, Araraquara, Brasil4

Received 20 February 2004/ Accepted 22 March 2004

Xylella fastidiosa is a phytopathogenic bacterium that causes serious diseases in a wide range of economically important crops. Despite extensive comparative analyses of genome sequences of Xylella pathogenic strains from different plant hosts, nonpathogenic strains have not been studied. In this report, we show that X. fastidiosa strain J1a12, associated with citrus variegated chlorosis (CVC), is nonpathogenic when injected into citrus and tobacco plants. Furthermore, a DNA microarray-based comparison of J1a12 with 9a5c, a CVC strain that is highly pathogenic and had its genome completely sequenced, revealed that 14 coding sequences of strain 9a5c are absent or highly divergent in strain J1a12. Among them, we found an arginase and a fimbrial adhesin precursor of type III pilus, which were confirmed to be absent in the nonpathogenic strain by PCR and DNA sequencing. The absence of arginase can be correlated to the inability of J1a12 to multiply in host plants. This enzyme has been recently shown to act as a bacterial survival mechanism by down-regulating host nitric oxide production. The lack of the adhesin precursor gene is in accordance with the less aggregated phenotype observed for J1a12 cells growing in vitro. Thus, the absence of both genes can be associated with the failure of the J1a12 strain to establish and spread in citrus and tobacco plants. These results provide the first detailed comparison between a nonpathogenic strain and a pathogenic strain of X. fastidiosa, constituting an important step towards understanding the molecular basis of the disease.

* Corresponding author. Mailing address: Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, Av. Prof. Lineu Prestes 748, 05508-900 São Paulo, SP, Brasil. Phone, fax, and e-mail for Suely L. Gomes: 5511 3091 3826, 5511 3815 5579, sulgomes{at}iq.usp.br. Phone fax, and e-mail for Aline M. da Silva: 5511 3091 2182, 5511 3815 5579, almsilva{at}iq.usp.br.

{dagger} Supplemental material for this article may be found at http://jb.asm.org/.

{ddagger} These authors contributed equally to this work.

§ Present address: Alellyx Applied Genomics, Campinas, SP, Brasil.

Journal of Bacteriology, August 2004, p. 5442-5449, Vol. 186, No. 16
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.16.5442-5449.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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