Xylella fastidiosa is a heterogenous gram-negative bacterial plant pathogen with a wide host range covering over 300 plant species. Since 2013, in Europe, the presence of the pathogen is increasing in a part of the Mediterranean area, but it causes in particular severe disease problems in olive orchards in the Southern part of Italy. Various subspecies of the pathogen were also diagnosed in natural outbreaks and intercepted ornamental plants in Europe, among them Olea europaea, Coffea arabica, and Nerium oleander. The host range of the pathogen can vary, depending on the subspecies and even the strain. The availability of fast and reliable diagnostic tools is indispensable in management strategies to control diseases caused by X. fastidiosa. To improve the reliability of the TaqMan assay, currently widely used in surveys, a triplex TaqMan assay was developed in which two specific and sensitive TaqMan assays, previously designed for X. fastidiosa, were combined with an internal control. The triplex assay exhibited the same diagnostic sensitivity as the simplex assays. In addition, the usefulness of a metagenomic approach using next-generation sequencing (NGS) was demonstrated, in which total DNA extracted from plant material was sequenced. DNA extracts from plant material free of X. fastidiosa, from artificially inoculated hosts plants or from naturally infected plants sampled in France, Spain, and Italy, or intercepted in Austria and the Netherlands, were analyzed for the presence of X. fastidiosa using the metagenomic approach. In all samples, even in samples with a low infection level, but not in the pathogen-free samples, DNA reads were detected specific for X. fastidiosa. In most cases, the pathogen could be identified to the subspecies level, and for one sample even the whole genome could be assembled and the sequence type could be determined. All results of NGS-analyzed samples were confirmed with the triplex TaqMan polymerase chain reaction and loop-mediated isothermal amplification.
Botryococcus braunii CCALA778 is a green microalga that can produce large amounts of extracellular carbohydrates and therefore is a potential host for industrial applications such as materials, food and pharmaceutical products. The downside of B. braunii is its slow growth and therefore, improvements on the biomass productivity or carbohydrate production will make this microalga more attractive for industrial exploitation. Microalgae grow naturally in the presence of bacteria and these can be beneficial or antagonistic. In outdoor cultivation systems, contamination by bacteria is common. The role or effects of bacteria present in B. braunii CCALA778 are not yet fully elucidated. We used UV-C treatment to reduce bacterial abundance in B. braunii CCALA778 cultures and 16S rRNA gene amplicon MiSeq sequencing for bacterial community analysis. The effect of the reduced amount of bacteria on biomass growth and production of extracellular carbohydrates was analysed. It is shown that UV-C treatment can reduce the bacterial population substantially without harming B. braunii. Bacteria removed by UV-C were antagonistic to B. braunii CCALA778 as in their absence production of biomass and extracellular carbohydrates was enhanced significantly. CCALA778 treated with UV-C accumulated 826 ± 61 mg L −1 of extracellular carbohydrates by day 15 compared with 422 ± 135 mg L −1 accumulated extracellular carbohydrates in the untreated culture.
Het beoogde weerbaarheidseffect betrof het zogenoemde systemic acquired resistance (SAR). De effecten van de behandelingen werden vastgesteld door bepaling van salicylzuur, van expressie van pathogenesis related (PR) proteins, en van de meeldauwontwikkeling.
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