Citrus is an economically important fruit crop that is severely afflicted by Asiatic citrus bacterial canker (CBC), a disease caused by the phytopathogen Xanthomonas citri subsp. citri (X. citri). To gain insight into the molecular epidemiology of CBC, 42 Xanthomonas isolates were collected from a range of Citrus spp. across 17 different orchards in Tucumán, Argentina and subjected to molecular, biochemical, and pathogenicity tests. Analysis of genome-specific X. citri markers and DNA polymorphisms based on repetitive elements-based polymerase chain reaction showed that all 42 isolates belonged to X. citri. Interestingly, pathogenicity tests showed that one isolate, which shares >90% genetic similarity to the reference strain X. citri T, has host range specificity. This new variant of X. citri subsp. citri, named X. citri A^T, which is deficient in xanthan production, induces an atypical, noncankerous chlorotic phenotype in Citrus limon and C. paradisi and weak cankerous lesions in C. aurantifolia and C. clementina leaves. In C. limon, suppression of canker development is concomitant with an oxidative burst; xanthan is not implicated in the phenotype induced by this interaction, suggesting that other bacterial factors would be involved in triggering the defense response.

Citrus canker, caused by Xanthomonas citri subsp. citri, and bacterial blight, caused by Xanthomonas citri subsp. malvacearum, results in substantial economic losses worldwide, and searching for new antibacterial agents is a critical challenge. In this study, regional isolates AE28 and RQ3 were obtained from characteristic lesions on Citrus limon and Gossypium hirsutum, respectively. Essential oils extracted by steam distillation from the fresh aerial parts of Pelargonium graveolens and Schinus molle exhibited complete (100%) inhibition of bacterial growth in vitro at a concentration of 1000 ppm, as determined by diffusion tests. To evaluate the potential of these essential oils for controlling Xanthomonas-induced diseases, in vivo assays were conducted on lemon leaves and cotton cotyledons inoculated with the regional AE28 and RQ3 strains. Two treatment approaches were tested: preventive application (24 h before inoculation) and curative application (24 h after inoculation). Preventive and curative treatments with P. graveolens essential oil significantly reduced citrus canker severity, whereas S. molle essential oil did not show a significant reduction compared to the control. In contrast, regardless of the treatment’s timing, both essential oils effectively reduced bacterial blight severity in cotton cotyledons by approximately 1.5-fold. Gas chromatography–mass spectrometry (GC-MS) analysis identified geraniol and citronellol as the major components of P. graveolens essential oil, while limonene and t-cadinol were predominant in S. molle. These findings highlight the promising potential of botanical products as bactericidal agents, warranting further research to optimize their application and efficacy.

Transcription activator-like effectors (TALEs) in plant-pathogenic Xanthomonas bacteria activate expression of plant genes and support infection or cause a resistance response. PthA4^AT is a TALE with a particularly short DNA-binding domain harboring only 7.5 repeats which triggers cell death in Nicotiana benthamiana; however, the genetic basis for this remains unknown. To identify possible target genes of PthA4^AT that mediate cell death in N. benthamiana, we exploited the modularity of TALEs to stepwise enhance their specificity and reduce potential target sites. Substitutions of individual repeats suggested that PthA4^AT-dependent cell death is sequence specific. Stepwise addition of repeats to the C-terminal or N-terminal end of the repeat region narrowed the sequence requirements in promoters of target genes. Transcriptome profiling and in silico target prediction allowed the isolation of two cell death inducer genes, which encode a patatin-like protein and a bifunctional monodehydroascorbate reductase/carbonic anhydrase protein. These two proteins are not linked to known TALE-dependent resistance genes. Our results show that the aberrant expression of different endogenous plant genes can cause a cell death reaction, which supports the hypothesis that TALE-dependent executor resistance genes can originate from various plant processes. Our strategy further demonstrates the use of TALEs to scan genomes for genes triggering cell death and other relevant phenotypes.