Leyva-López, Norma Elena

The increasing impact of phloem-restricted bacteria on economically important crops has led to renewed interest in understanding the pathogenesis at the genomic and histological levels of these diseases. The genus Candidatus Liberibacter is associated with economically devastating diseases, highlighting Candidatus Liberibacter asiaticus (CLas) and Candidatus Liberibacter solanaceraum (CLso) in citrus and vegetables. Plant-pathogen interaction studies are limited due to the non-culturable nature of the pathogenic species of the genus Ca. Liberibacter, as well as the lack of experimental models. The objective of this work was to implement a model for the study of bacteria of the genus Candidatus Liberibacter, using tomato-CLso as an experimental model. Through a time course analysis of the tomato-CLso infective process, it was determined that there is a direct correlation between the bacterial load and symptom development, differentiated into early, intermediate and late stages. Additionally, a comparative transcriptional analysis of antibacterial defense marker genes (PR-P1, miR160, and miR393) determined that the response of these genes in the tomato-CLso and lime-CLas pathosystems are equivalent. Our results demonstrate that the tomato-CLso system represents an important model for the study of the interaction between plants and non-cultivable phloem restricted bacteria.

Candidatus Liberibacter asiaticus (CLas) se ha dispersado rápidamente a las regiones productoras de cítricos en México, ocasionando impactos económicos y sociales diferenciados. En México, los principales avances en investigación se han centrado en el manejo del vector, Diaphorina citri y en estudios de la interacción de CLas con limón mexicano. Sin embargo, la variabilidad genética de la bacteria en regiones productoras de México se ha estudiado poco. Por lo que, en el presente estudio se analizó la variación de la población mexicana de CLas con procedimientos basados en PCR, secuenciación y electroforesis capilar a través de la detección del número de repeticiones en tándem (TRN) con dos marcadores STR, AGACACA y TACAGAA, el primero localizado en locus genómicos CLIBASIA_01645 y adenosina deaminasas. La distribución de los TRNs en la población mexicana de CLas en ambos loci mostraron variaciones con las poblaciones previamente reportadas en Florida, China y Brasil. Además, en la población mexicana se encontraron tres haplotipos, HA, HB y HC, siendo el primer reporte del haplotipo HC en las regiones citrícolas en México. Interesantemente, las subpoblaciones de CLas del Occidente y Noroeste son diferentes a la del Sureste.

Huanglongbing (HLB) is one of the most destructive diseases threatening citriculture worldwide. This disease has been associated with α-proteobacteria species, namely Candidatus Liberibacter. Due to the unculturable nature of the causal agent, it has been difficult to mitigate the disease, and nowadays a cure is not available. MicroRNAs (miRNAs) are key regulators of gene expression, playing an essential role in abiotic and biotic stress in plants including antibacterial responses. However, knowledge derived from non-model systems including Candidatus Liberibacter asiaticus (CLas)-citrus pathosystem remains largely unknown. In this study, small RNA profiles from Mexican lime (Citrus aurantifolia) plants infected with CLas at asymptomatic and symptomatic stages were generated by sRNA-Seq, and miRNAs were obtained with ShortStack software. A total of 46 miRNAs, including 29 known miRNAs and 17 novel miRNAs, were identified in Mexican lime. Among them, six miRNAs were deregulated in the asymptomatic stage, highlighting the up regulation of two new miRNAs. Meanwhile, eight miRNAs were differentially expressed in the symptomatic stage of the disease. The target genes of miRNAs were related to protein modification, transcription factors, and enzyme-coding genes. Our results provide new insights into miRNA-mediated regulation in C. aurantifolia in response to CLas infection. This information will be useful to understand molecular mechanisms behind the defense and pathogenesis of HLB.

Nowadays, Huanglongbing (HLB) disease, associated with Candidatus Liberibacter asiaticus (CLas), seriously affects citriculture worldwide, and no cure is currently available. Transcriptomic analysis of host–pathogen interaction is the first step to understand the molecular landscape of a disease. Previous works have reported the transcriptome profiling in response to HLB in different susceptible citrus species; however, similar studies in tolerant citrus species, including Mexican lime, are limited. In this work, we have obtained an RNA-seq-based differential expression profile of Mexican lime plants challenged against CLas infection, at both asymptomatic and symptomatic stages. Typical HLB-responsive differentially expressed genes (DEGs) are involved in photosynthesis, secondary metabolism, and phytohormone homeostasis. Enrichment of DEGs associated with biotic response showed that genes related to cell wall, secondary metabolism, transcription factors, signaling, and redox reactions could play a role in the tolerance of Mexican lime against CLas infection. Interestingly, despite some concordance observed between transcriptional responses of different tolerant citrus species, a subset of DEGs appeared to be species-specific. Our data highlights the importance of studying the host response during HLB disease using as model tolerant citrus species, in order to design new and opportune diagnostic and management methods.

El Huanglongbing representa un gran reto para su control, por lo que se requiere de alternativas efectivas a la aplicación de insecticidas al insecto vector. La inducción de resistencia sistémica es una alternativa para disminuir el progreso de la enfermedad en árboles infectados. El objetivo de este trabajo fue evaluar el efecto de inductores de resistencia sistémica sobre la concentración de <em>?Candidatus</em> Liberibacter asiaticus? (CLas) en árboles de limón mexicano (Lm) en condiciones de invernadero. Los inductores de resistencia utilizados fueron ácido salicílico (T1), <em>Azospirillum brasilense</em> Cd (T2) y quitosano (T3), aplicados cada 20 días durante ocho meses. La cuantificación de CLas fue mediante qPCR. La concentración de CLas se determinó a los 1, 2, 5 y 8 meses pos-tratamiento. A los 8 meses pos-tratamiento, T1 y T3 no presentaron diferencias postratamiento en la reducción postratamiento de la concentración de CLas con respecto al control sin inductor (T4) (6.5×103 células bacterianas/100 ng de ADN), mientras que T2 disminuyó casi tres veces la concentración de CLas con respecto al control. La inoculación directa al suelo de <em>A. brasilense</em> Cd mostró un efecto significativo en la reducción de la concentración de CLas en los árboles de Lm en condiciones de invernadero.