A multiplex real-time PCR method based on fluorescent TaqMan® probes was developed for the simultaneous detection of the tomato pathogenic bacteria Clavibacter michiganensis subsp. michiganensis, Pseudomonas syringae pv. tomato and bacterial spot-causing xanthomonads. The specificity of the multiplex assay was validated on 44 bacterial strains, including 32 target pathogen strains as well as closely related species and nontarget tomato pathogenic bacteria. The designed multiplex real-time PCR showed high sensitivity when positive amplification was observed for one pg of bacterial DNA in the cases of Clavibacter michiganensis subsp. michiganensis and Pseudomonas syringae pv. tomato bacteria and 100 pg for bacterial spot-causing xanthomonads. The reliability of the developed multiplex real-time PCR assay for in planta detection was verified by recognition of the target pathogens in 18 tomato plants artificially inoculated by each of the target bacteria and tomato samples from production greenhouses.
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- Actinobacteria genetika izolace a purifikace fyziologie MeSH
- kvantitativní polymerázová řetězová reakce * MeSH
- prostředí kontrolované MeSH
- Pseudomonas syringae genetika izolace a purifikace fyziologie MeSH
- Solanum lycopersicum růst a vývoj mikrobiologie MeSH
- Xanthomonas genetika izolace a purifikace fyziologie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Diaporthe species are important pathogens, saprobes, and endophytes on grapevines. Several species are known, either as agents of pre- or post-harvest infections, as causal agents of many relevant diseases, including swelling arm, trunk cankers, leaf spots, root and fruit rots, wilts, and cane bleaching. A growing body of evidence exists that a class of small non-coding endogenous RNAs, known as microRNAs (miRNAs), play an important role in post-transcriptional gene regulation, during plant development and responses to biotic and abiotic stresses. In this study, we explored differentially expressed miRNAs in response to Diaporthe eres and Diaporthe bohemiae infection in Vitis vinifera cv. Chardonnay under in vitro conditions. We used computational methods to predict putative miRNA targets in order to explore the involvement of possible pathogen response pathways. We identified 136 known and 41 new miRNA sequence variants, likely generated through post-transcriptional modifications. In the Diaporthe eres treatment, 61 known and 17 new miRNAs were identified while in the Diaporthe bohemiae treatment, 101 known and 21 new miRNAs were revealed. Our results contribute to further understanding the role miRNAs play during plant pathogenesis, which is possibly crucial in understanding disease symptom development in grapevines infected by D. eres and D. bohemiae.
"Candidatus Phytoplasma prunorum" (CPp) is a highly destructive phytopathogenic agent in many stone fruit-growing regions in Europe and the surrounding countries. In this work, we focused on documenting entire bacterial community in the phloem tissues of 60 stone fruit trees. Nested PCR and two real-time PCR assays were used to select CPp-positive (group A) and CPp-negative samples (group B). Afterwards, high-throughput amplicon sequencing was performed to assess bacterial community compositions in phloem tissues. The bacterial composition in phloem tissue consisted of 118 distinct genera, represented mainly by Pseudomonas, Acinetobacter, Methylobacterium, Sphingomonas, and Rhizobium. Statistics showed that CPp influenced the bacterial composition of infected plants (group A) and that the bacterial community depended on the geographical origin of the sample. This is the first work focusing on an analysis of the influence of CPp on the bacteria coexisting in the phloem tissues of stone fruit trees.
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- financování organizované MeSH
- Publikační typ
- abstrakty MeSH
- MeSH
- bronchoalveolární laváž MeSH
- experimenty na zvířatech MeSH
- finanční podpora výzkumu jako téma MeSH
- financování organizované MeSH
- keramika toxicita MeSH
- látky znečišťující vzduch v pracovním prostředí MeSH
- modely nemocí na zvířatech MeSH
- plicní nemoci MeSH
- potkani Wistar MeSH
- silikáty hliníku toxicita MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- MeSH
- financování organizované MeSH
- Publikační typ
- abstrakty MeSH
- MeSH
- alveolární makrofágy MeSH
- amositový azbest * analýza izolace a purifikace toxicita MeSH
- bronchoalveolární laváž MeSH
- cytotoxicita imunologická * MeSH
- kathepsin D analýza MeSH
- kouření * škodlivé účinky MeSH
- kyselá fosfatasa analýza MeSH
- L-laktátdehydrogenasa analýza MeSH
- longitudinální studie * MeSH
- minerální vlákna * analýza toxicita MeSH
- náhodné rozdělení MeSH
- potkani Wistar MeSH
- pracovní expozice MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- práce podpořená grantem MeSH
