New genes of Xanthomonas citri subsp. citri involved in pathogenesis and adaptation revealed by a transposon-based mutant library

Autor: Julio Cezar Franco de Oliveira, Joice Bissoloti Brigati, Maria Inês Tiraboschi Ferro, Ana Cristina Simões e Silva, Juliana Dezajacomo, Marcelo Luiz de Laia, Cristiano Barbalho Ferreira, Jesus Aparecido Ferro, Leandro Marcio Moreira
Přispěvatelé: Universidade Estadual Paulista (Unesp), Universidade de São Paulo (USP), Alellyx Appl Genom, Universidade Federal de São Paulo (UNIFESP), Univ Estadual Paulista
Jazyk: angličtina
Předmět:
Zdroj: Repositório Institucional da UFOP
Universidade Federal de Ouro Preto (UFOP)
instacron:UFOP
BMC Microbiology
Web of Science
Repositório Institucional da UNESP
Universidade Estadual Paulista (UNESP)
instacron:UNESP
BMC Microbiology, Vol 9, Iss 1, p 12 (2009)
Repositório Institucional da UNIFESP
Universidade Federal de São Paulo (UNIFESP)
instacron:UNIFESP
ISSN: 1471-2180
DOI: 10.1186/1471-2180-9-12
Popis: Made available in DSpace on 2013-08-28T14:10:11Z (GMT). No. of bitstreams: 1 WOS000264158500001.pdf: 2073598 bytes, checksum: 1b34ea00cdecd061a7e6523efc9fda61 (MD5) Made available in DSpace on 2013-09-30T18:00:21Z (GMT). No. of bitstreams: 2 WOS000264158500001.pdf: 2073598 bytes, checksum: 1b34ea00cdecd061a7e6523efc9fda61 (MD5) WOS000264158500001.pdf.txt: 78494 bytes, checksum: 67bc62d4f386ee061ca41da6854d57ee (MD5) Previous issue date: 2009-01-16 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T13:17:43Z No. of bitstreams: 2 WOS000264158500001.pdf: 2073598 bytes, checksum: 1b34ea00cdecd061a7e6523efc9fda61 (MD5) WOS000264158500001.pdf.txt: 78494 bytes, checksum: 67bc62d4f386ee061ca41da6854d57ee (MD5) Made available in DSpace on 2014-05-20T13:17:43Z (GMT). No. of bitstreams: 2 WOS000264158500001.pdf: 2073598 bytes, checksum: 1b34ea00cdecd061a7e6523efc9fda61 (MD5) WOS000264158500001.pdf.txt: 78494 bytes, checksum: 67bc62d4f386ee061ca41da6854d57ee (MD5) Previous issue date: 2009-01-16 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Fundo de Defesa da Citricultura (FUNDECITRUS) Background: Citrus canker is a disease caused by the phytopathogens Xanthomonas citri subsp. citri, Xanthomonas fuscans subsp. aurantifolli and Xanthomonas alfalfae subsp. citrumelonis. The first of the three species, which causes citrus bacterial canker type A, is the most widely spread and severe, attacking all citrus species. In Brazil, this species is the most important, being found in practically all areas where citrus canker has been detected. Like most phytobacterioses, there is no efficient way to control citrus canker. Considering the importance of the disease worldwide, investigation is needed to accurately detect which genes are related to the pathogen-host adaptation process and which are associated with pathogenesis.Results: Through transposon insertion mutagenesis, 10,000 mutants of Xanthomonas citri subsp. citri strain 306 (Xcc) were obtained, and 3,300 were inoculated in Rangpur lime (Citrus limonia) leaves. Their ability to cause citrus canker was analyzed every 3 days until 21 days after inoculation; a set of 44 mutants showed altered virulence, with 8 presenting a complete loss of causing citrus canker symptoms. Sequencing of the insertion site in all 44 mutants revealed that 35 different ORFs were hit, since some ORFs were hit in more than one mutant, with mutants for the same ORF presenting the same phenotype. An analysis of these ORFs showed that some encoded genes were previously known as related to pathogenicity in phytobacteria and, more interestingly, revealed new genes never implicated with Xanthomonas pathogenicity before, including hypothetical ORFs. Among the 8 mutants with no canker symptoms are the hrpB4 and hrpX genes, two genes that belong to type III secretion system (TTSS), two hypothetical ORFS and, surprisingly, the htrA gene, a gene reported as involved with the virulence process in animal-pathogenic bacteria but not described as involved in phytobacteria virulence. Nucleic acid hybridization using labeled cDNA probes showed that some of the mutated genes are differentially expressed when the bacterium is grown in citrus leaves. Finally, comparative genomic analysis revealed that 5 mutated ORFs are in new putative pathogenicity islands.Conclusion: The identification of these new genes related with Xcc infection and virulence is a great step towards the understanding of plant-pathogen interactions and could allow the development of strategies to control citrus canker. Univ Estadual Paulista, UNESP, Dept Technol, BR-14884900 Jaboticabal, SP, Brazil Univ São Paulo, Inst Quim, Dept Bioquim, BR-05509900 São Paulo, Brazil Alellyx Appl Genom, BR-13069380 Campinas, SP, Brazil Univ Fed São Paulo, Dept Ciencias Biol, BR-09972270 Diadema, SP, Brazil Univ Estadual Paulista, UNESP, Dept Technol, BR-14884900 Jaboticabal, SP, Brazil FAPESP: 04/02006-7
Databáze: OpenAIRE
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