Visualization of the impatiens downy mildew pathogen using fluorescence in situ hybridization (FISH)
| العنوان: | Visualization of the impatiens downy mildew pathogen using fluorescence in situ hybridization (FISH) |
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| المؤلفون: | Catalina Salgado-Salazar, Jo Anne Crouch, Gary R. Bauchan, Emma C. Wallace |
| المصدر: | Plant Methods, Vol 14, Iss 1, Pp 1-9 (2018) Plant Methods |
| بيانات النشر: | BMC, 2018. |
| سنة النشر: | 2018 |
| مصطلحات موضوعية: | 0106 biological sciences, 0301 basic medicine, Hypha, Plant Science, lcsh:Plant culture, 01 natural sciences, Microbiology, 03 medical and health sciences, Plasmopara obducens, Genetics, medicine, Oomycota, lcsh:SB1-1110, Natural environment, lcsh:QH301-705.5, Oomycete, biology, medicine.diagnostic_test, Peronosporales, Methodology, Oospores, biology.organism_classification, Impatiens walleriana, 030104 developmental biology, lcsh:Biology (General), Downy mildew, Impatiens, Oligomer restriction, 010606 plant biology & botany, Biotechnology, Fluorescence in situ hybridization |
| الوصف: | Background Plasmopara obducens is the biotrophic oomycete responsible for impatiens downy mildew, a destructive disease of Impatiens that causes high crop loss. Currently, there are no available methods for the microscopic detection of P. obducens from leaves of impatiens, which may be contributing to the spread of the disease. Fluorescence in situ hybridization (FISH) is a sensitive and robust method that uses sequence-specific, fluorescence-labeled oligonucleotide probes to detect target organisms from the environment. To study this important pathogen, we developed and standardized a FISH technique for the visualization of P. obducens from Impatiens walleriana tissues using a species-specific 24-mer oligonucleotide probe designed to target a region of the rRNA internal transcribed spacer 2 (ITS2). Results Since P. obducens cannot be propagated in vitro, we developed a custom E. coli expression vector that transcribes the P. obducens rRNA-ITS target sequence (clone-FISH) for use as a control and to optimize hybridization conditions. The FISH assay could detect P. obducens sporangiophores, sporangia and oospores, and hyphae from naturally infected I. walleriana leaves and stems. Cross-reactivity was not observed from plant tissue, and the assay did not react when applied to E. coli with self-ligated plasmids and non-target oomycete species. Conclusions This FISH protocol may provide a valuable tool for the study of this disease and could potentially be used to improve early monitoring of P. obducens, substantially reducing the persistence and spread of this destructive plant pathogen. |
| اللغة: | English |
| تدمد: | 1746-4811 |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::1c60e70ad1ae3b21e17471a7d1b92141Test http://link.springer.com/article/10.1186/s13007-018-0362-zTest |
| حقوق: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....1c60e70ad1ae3b21e17471a7d1b92141 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 17464811 |
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