Project description:microRNAs can play a crucial role in stress response in plants, including biotic stress. Some miRNAs are known to respond to bacterial infection. This work has addressed the role of miRNAs in Manihot esculenta (cassava)-Xanthomonas axonopodis pv. manihotis (Xam) interaction. Illumina sequencing was used for analyzing small RNA libraries from cassava tissue infected and non-infected with Xam. Cassava variety MBRA685 (resistant to Xam-CIO151) Six-week-old plants were inoculated with 36h-old cultures of the aggressive Xanthomonas axonopodis pv. manihotis strain CIO151 in both leaves and stems.
Project description:microRNAs can play a crucial role in stress response in plants, including biotic stress. Some miRNAs are known to respond to bacterial infection. This work has addressed the role of miRNAs in Manihot esculenta (cassava)-Xanthomonas axonopodis pv. manihotis (Xam) interaction. Illumina sequencing was used for analyzing small RNA libraries from cassava tissue infected and non-infected with Xam. Cassava variety MBRA685 (resistant to Xam-CIO151) Six-week-old plants were inoculated with 36h-old cultures of the aggressive Xanthomonas axonopodis pv. manihotis strain CIO151 in both leaves and stems. Leaves were inoculated by piercing six holes in the mesophyll and placing a 5µL drop of a liquid Xam-MgCl2 culture calibrated at OD600nm = 0.002 (1 x108cfu/ml). Two leaflets per leaf and three leaves per plant were inoculated. Stems were inoculated by puncture in the stems as described previously (24). At least three plants per collection time were inoculated. Leaves and stems were collected from inoculated plants (0 hours post inoculation -hpi, 6hpi, 24hpi, 2 days post-inoculation -dpi, 5dpi, 7dpi and 15dpi) and non-inoculated plants. RNA extractions were made using a LiCl-acid phenol:chloroform method.
Project description:Xanthomonas axonopodis pv. manihotis (Xam) is a gram negative bacterium causing Cassava Bacterial Blight (CBB), an important limitation for cassava production. The genetic bases underlying cassava resistance and susceptibility to different Xam strains are currently unknown. To identify genes and pathways important for the interaction, we used RNA-seq data to study transcriptomic changes in cassava plants inoculated with the non-pathogenic Xam strain, (ORST4) and a pathogenic strain, ORST4 transformed with the TAL effector TALE1Xam (ORST4+TALE1Xam). This analysis revealed that transcriptomic responses to both strains were very similar and were dominated by the induction of genes related to photosynthesis and phenylpropanoid biosynthesis and the down-regulation of genes related to jasmonic acid signaling, features possibly related to defense responses. Among the genes induced exclusively in cassava plants inoculated with ORST4 + TALE1Xam we found one gene containing a predicted binding site for TALE1Xam in its promoter region. This gene encodes for a Heat Shock Transcription Factor B3 (HsfB3) and likely acts a transcriptional repressor. HsfB3 may constitute a new type of susceptibility gene activated by a TAL effector that manages to be sufficient for symptom development without suppressing defense responses in the plant. mRNA of Cassava stems inoculated with a non-pathogenic (ORST4) and pathogenic (+TALE1Xam) strain of Xanthomonas axonopodis pv. Manihotis, tissues collected at 0, 5 and7 days post-inoculation, 2 technical replicates used
Project description:The goal of this study was to identify signaling processes associated with infection and recovery of leafy spurge inoculated with Xanthomonas axonopodis pv. manihotis (Xam).
Project description:The goal of this study was to identify signaling processes associated with infection and recovery of leafy spurge inoculated with Xanthomonas axonopodis pv. manihotis (Xam). Two dye-swap technical replicates for each of 3 biological replicates for each treatment were analyzed. A time course analysis of Xam infected and mock inoculated leafy spurge was performed. Time points were 1, 7 and 21 days after inoculation (or mock inoculation). Thus, 36 labeled samples were hybridized in a 2 dye rolling circle hybridization scheme to 18 arrays.
Project description:Xanthomonas axonopodis pv. manihotis (Xam) is a gram negative bacterium causing Cassava Bacterial Blight (CBB), an important limitation for cassava production. The genetic bases underlying cassava resistance and susceptibility to different Xam strains are currently unknown. To identify genes and pathways important for the interaction, we used RNA-seq data to study transcriptomic changes in cassava plants inoculated with the non-pathogenic Xam strain, (ORST4) and a pathogenic strain, ORST4 transformed with the TAL effector TALE1Xam (ORST4+TALE1Xam). This analysis revealed that transcriptomic responses to both strains were very similar and were dominated by the induction of genes related to photosynthesis and phenylpropanoid biosynthesis and the down-regulation of genes related to jasmonic acid signaling, features possibly related to defense responses. Among the genes induced exclusively in cassava plants inoculated with ORST4 + TALE1Xam we found one gene containing a predicted binding site for TALE1Xam in its promoter region. This gene encodes for a Heat Shock Transcription Factor B3 (HsfB3) and likely acts a transcriptional repressor. HsfB3 may constitute a new type of susceptibility gene activated by a TAL effector that manages to be sufficient for symptom development without suppressing defense responses in the plant.
Project description:Pathogens rely on expression of host susceptibility (S) genes to promote infection and disease. DNA methylation is an epigenetic modification that affects gene expression. Theoretically, blocking access to S genes through targeted methylation could increase disease resistance. Xanthomonas axonopodis pv. manihotis, the causal agent of cassava bacterial blight (CBB), uses transcription activator-like20 (TAL20) to induce expression of the S gene MeSWEET10a. We directed methylation to the TAL20 effector binding element within the MeSWEET10a promoter using a synthetic zinc-finger DNA binding domain fused to a component of the RNA-directed DNA methylation pathway. We demonstrate that this methylation prevents TAL20 binding, blocks transcriptional activation of MeSWEET10a and that these plants display increased resistance to CBB. This work establishes epigenome editing as a new strategy for crop improvement.
Project description:Manihot esculenta Transcriptome (TAL Effector Repertoires of Strains of Xanthomonas phaseoli pv. manihotis in Commercial Cassava Crops Reveal High Diversity at the Country Scale)