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Franco Ortega, S.

Publications and source records attributed to Franco Ortega, S..

3 recordsLinked to original sources

Detecting transcriptional responses and comparing the virulence of Pseudomonas aeruginosa cystic fibrosis isolates in a mung bean model

A mung bean infection model has previously been shown to differentiate between non-virulent and virulent Pseudomonas aeruginosa bacteria. However, it remains unclear how plant and bacteria adjust their gene expression during infection and whether the mung bean model can be used to compare the virulence of clinical cystic fibrosis (CF) P. aeruginosa lung isolates. Here, we first explored temporal transcriptomics of P. aeruginosa PAO1 and mung bean during an infection. We found that bacterial gene expression followed temporal changes, with an increase in the expression of O-antigen biosynthetic genes, chemotaxis, phosphate intake and phenazine production. Mung bean responded by upregulating genes associated with defence mechanisms and downregulating genes involved in the plant development. From the PAO1 perspective, the core-transcriptomic responses in the mung bean were similar to its responses previously observed in wound and excision and in in vitro media and sputum models, while differed from those observed in the bronchial cell model. Furthermore, we used the mung bean to assess the virulence of 119 clinical P. aeruginosa CF strains originating from the Copenhagen CF clinic. By quantifying bacterial virulence as a reduction in root and shoot growth and weight of the seeds, we found that CF strains isolated at later compared to early stages of lung infections showed higher virulence. This difference corresponded with the higher number of immune modulation-associated virulence genes and lower number of motility and effector genes, present in the genomes of late compared to early isolated CF strains. IMPORTANCEOur results demonstrate that based on PAO1 transcriptional profile, the mung bean model is similar to in vitro and wound infection models but differs from cell and bronchial models. Moreover, the mung bean model can detect virulence differences between clinical P. aeruginosa CF strains, making it a potentially useful high-throughput in vivo model for bacterial virulence screening.

microbiology↗

Assembly and annotation of Solanum dulcamara and Solanum nigrum plant genomes, two nightshades with different susceptibilities to Ralstonia solanacearum

To understand why close wild plant relatives of crops, such as Solanum dulcamara, are resistant to Ralstonia solanacearum we need genetic resources to perform comparative studies and identify key genes and pathways. We de-novo assembled and annotated the genome of resistant S. dulcamara and susceptible Solanum nigrum plants using a hybrid approach including Oxford Nanopore Technologies and Illumina sequencing. Comparative genomic analysis was then performed to find differences between the genome of S. dulcamara and other susceptible Solanaceous species including potato, tomato, aubergine, and S. nigrum and one susceptible and one resistant S. americanum accession. We identified genes associated with auxin-transport only in S. dulcamara and a collection of pattern recognition receptors (PRRs) was identified in orthogroups only found in plant species with resistant/tolerant phenotype, suggesting novel plant receptors in these accessions that may improve recognition of pathogen-associated molecular patterns (PAMPs) associated with R. solanacearum. We also identified differences in methylation frequency across the gene bodies in both species, which may be associated with epigenetic regulation of resistance. Future work should assess the functional role of these PRRs during bacterial wilt development to determine if they could offer potential novel targets for breeding improved wilt resistance.

plant biology↗

Fraxinus excelsior updated long-read genome reveals the importance of MADS-box genes in tolerance mechanisms against ash dieback

Ash dieback caused by the fungus Hymenoscyphus fraxineus has devastated the European ash tree population since it arrived in Europe in 1992. Great effort has been put into breeding programmes to increase the genetic diversity of ash trees and find heritable genetic markers associated with resistance, or tolerance mechanisms, to ash dieback. To facilitate identification of molecular markers, we used Oxford Nanopore Technologies combined with Illumina sequencing to obtain an accurate and contiguous ash genome. We used this genome to reanalyse transcriptome data from a Danish ash panel of 182 tree accessions. Using associative transcriptomics, we identified 175 gene expression markers (GEMs), including 11 genes annotated as dormancy MADS-box transcription factors which are associated with ash bud dormancy, flowering and senescence. We hypothesize that tolerant trees both break dormancy earlier in the year by increasing the expression of flowering-related SOC1 MADS-box and reducing the expression of SVP-like MADS-box, whilst also accelerating senescence by increasing the expression of JOINTLESS MADS-box genes. DNA methylation differences in the promoters of MADS-box genes between one tolerant and one susceptible tree indicate potential epigenetic regulation of these traits. Article SummaryAsh dieback has devastated European ash tree populations. To aid in breeding programmes focused on finding solutions against this pathogen, we have assembled a new ash genome. This new genome helped us to identify genes related to tree biological life cycles, expressed differently in tolerant and susceptible trees. For the first time, we have also discovered that susceptible and tolerant trees showed different DNA methylation frequencies in those genes, suggesting epigenetic regulation. DNA methylation can turn on/off gene expression without changing the DNA sequence. These genes, and their regulatory elements, are ideal targets during breeding programmes combating this pathogen.

plant biology↗