RESUMEN
The type III effector AvrXv3 from Xanthomonas campestris pv. vesicatoria (Xcv) elicits a resistance response in the tomato line Hawaii 7981. To test whether similar genes participate in responses triggered by recognition of different avirulence proteins, we examined the effect of AvrXv3 expression on the plant transcriptome as compared to that of other avirulence proteins. By microarray analysis we monitored expression of approximately 8,600 tomato genes upon inoculation with isogenic Xcv strains differing only by the avrXv3 gene. Changes in transcript levels of 139 genes were observed within 8 h, and a massive shift in expression of 1,294 genes was detected at 12 h. Recognition of AvrXv3 modulated a large number of genes encoding transcription factors and signaling components. In addition, genes involved in defense and stress responses, lipid metabolism, protein degradation, and secondary metabolism were mainly up-regulated. Conversely, genes related to photosynthesis and protein synthesis were generally down-regulated. Many novel genes encoding proteins of unknown function were also identified. A comparison between AvrXv3-modulated genes and those differentially expressed in tomato plants recognizing other bacterial effectors revealed partial overlap and similar distribution in functional classes. The identification of tomato genes modulated by AvrXv3 expression paves the way for dissecting defense networks activated by recognition of this effector in resistant plants.
Asunto(s)
Solanum lycopersicum/genética , Solanum lycopersicum/microbiología , Xanthomonas campestris/patogenicidad , Proteínas Bacterianas/genética , Perfilación de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Genes Bacterianos , Genes de Plantas , Genómica , Solanum lycopersicum/metabolismo , Modelos Biológicos , Análisis de Secuencia por Matrices de Oligonucleótidos , Enfermedades de las Plantas/genética , Enfermedades de las Plantas/microbiología , Transactivadores/genética , Xanthomonas campestris/genéticaRESUMEN
The Xanthomonas campestris pv. vesicatoria avirulence gene AvrRxv specifies resistance on the tomato line Hawaii 7998 by interacting with three nondominant plant resistance genes. AvrRxv molecular properties that impinge on its avirulence activity were characterized and transcriptional changes caused by AvrRxv expression in resistant tomato plants were extensively examined. AvrRxv localized predominantly to the cytoplasm and possibly in association with plasma and nuclear membranes in both resistant and susceptible tomato plants. The AvrRxv cysteine protease catalytic core was found to be essential for host recognition, because introduction of mutations in this domain affected the ability of AvrRxv to elicit a hypersensitive response and the inhibition of bacterial growth in resistant plants. In addition, expression profiles were analyzed for approximately 8,600 tomato genes in resistant plants challenged with X. campestris pv. vesicatoria strains expressing wild-type AvrRxv or a catalytic core AvrRxv mutant. In all, 420 genes were identified as differentially modulated by the expression of a functional AvrRxv, including over 15 functional classes of proteins and a large number of transcription factors and signaling components. Findings of this study allow the development of new hypotheses about the molecular basis of recognition between AvrRxv and the corresponding resistance proteins, and set the stage for the dissection of signaling and cellular responses triggered in tomato plants by this avirulence factor.
Asunto(s)
Proteínas Bacterianas/genética , Expresión Génica , Solanum lycopersicum/genética , Transcripción Genética , Xanthomonas/genética , Proteínas Bacterianas/metabolismo , Regulación hacia Abajo , Perfilación de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Inmunidad Innata , Solanum lycopersicum/metabolismo , Solanum lycopersicum/microbiología , Mutación , Enfermedades de las Plantas/genética , Enfermedades de las Plantas/microbiología , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Regulación hacia Arriba , Virulencia , Xanthomonas/metabolismo , Xanthomonas/patogenicidadRESUMEN
The gram-negative bacterium Xanthomonas campestris pv. vesicatoria is the causal agent of spot disease in tomato and pepper. Plants of the tomato line Hawaii 7981 are resistant to race T3 of X. campestris pv. vesicatoria expressing the type III effector protein AvrXv3 and develop a typical hypersensitive response upon bacterial challenge. A combination of suppression subtractive hybridization and microarray analysis identified a large set of cDNAs that are induced or repressed during the resistance response of Hawaii 7981 plants to X. campestris pv. vesicatoria T3 bacteria. Sequence analysis of the isolated cDNAs revealed that they correspond to 426 nonredundant genes, which were designated as XRE (Xanthomonas-regulated) genes and were classified into more than 20 functional classes. The largest functional groups contain genes involved in defense, stress responses, protein synthesis, signaling, and photosynthesis. Analysis of XRE expression kinetics during the tomato resistance response to X. campestris pv. vesicatoria T3 revealed six clusters of genes with coordinate expression. In addition, by using isogenic X. campestris pv. vesicatoria T2 strains differing only by the avrXv3 avirulence gene, we found that 77% of the identified XRE genes were directly modulated by expression of the AvrXv3 effector protein. Interestingly, 64% of the XRE genes were also induced in tomato during an incompatible interaction with an avirulent strain of Pseudomonas syringae pv. tomato. The identification and expression analysis of X. campestris pv. vesicatoria T3-modulated genes, which may be involved in the control or in the execution of plant defense responses, set the stage for the dissection of signaling and cellular responses activated in tomato plants during the onset of spot disease resistance.