<em>In situ</em> immunofluorescence localization: A method for rapid detection of <em>Beauveria</em> spp. in the rhizosphere of <em>Quercus robur</em> saplings
For biological control of plant pests, e.g. cockchafer grubs, in the rhizosphere of oak, apple or pine trees, entomopathogenic Beauveria spp. are increasingly applied. For successful use, it is important to monitor the spread and persistence of the inoculated fungi, both qualitatively and quantita...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | deu |
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Julius Kühn-Institut
2019-07-01
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| Series: | Journal für Kulturpflanzen |
| Subjects: | |
| Online Access: | https://ojs.openagrar.de/index.php/Kulturpflanzenjournal/article/view/13009 |
| _version_ | 1819279723673419776 |
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| author | Marta Matek Cornelia I. Ullrich Frank Rabenstein Eckhard Koch Regina G. Kleespies |
| author_facet | Marta Matek Cornelia I. Ullrich Frank Rabenstein Eckhard Koch Regina G. Kleespies |
| author_sort | Marta Matek |
| collection | DOAJ |
| description | For biological control of plant pests, e.g. cockchafer grubs, in the rhizosphere of oak, apple or pine trees, entomopathogenic Beauveria spp. are increasingly applied. For successful use, it is important to monitor the spread and persistence of the inoculated fungi, both qualitatively and quantitatively. The determination of both parameters by plating on selective nutrient media or by molecular methods such as PCR of soil samples are quite laborious and often do not yield satisfactory results. Therefore, the aim of the present study was to develop a specific in situ method using immunofluorescence labelling of Beauveria spp. growing on young fine roots of three-year old oak saplings. All fine roots investigated were covered with a dense net of soil rhizosphere fungi, as visualized by staining with the nonspecific dye blankophor. On non-inoculated roots, polyclonal Beauveria antibodies did not label any of these naturally growing fungi. Only samples of roots inoculated with Beauveria brongniartii displayed specific labelling up to ten months after inoculation. Whereas the natural rhizosphere fungi were detected growing in the intercellular space of the root cortex in an ectomycorrhiza-like manner up to the endodermis, hyphae of the inoculated B. brongniartii were never seen within the root tissue but only growing on the surface of the rhizodermis. These observations indicate that B. brongniartii does not grow endophytically, and that the method used allows to discriminate B. brongniartii from the resident fungal flora in the oak tree rhizosphere. Detection by immunofluorescence labelling employed in the current study may be a useful tool to follow B. brongniartii in experiments aimed at establishing the entomopathogen in the rhizosphere and to monitor its fate in long-term control of entomopathogens. |
| first_indexed | 2024-12-24T00:32:26Z |
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| id | doaj.art-86c7d7344096441d85d98da74a978fe5 |
| institution | Directory Open Access Journal |
| issn | 1867-0911 1867-0938 |
| language | deu |
| last_indexed | 2024-12-24T00:32:26Z |
| publishDate | 2019-07-01 |
| publisher | Julius Kühn-Institut |
| record_format | Article |
| series | Journal für Kulturpflanzen |
| spelling | doaj.art-86c7d7344096441d85d98da74a978fe52022-12-21T17:24:12ZdeuJulius Kühn-InstitutJournal für Kulturpflanzen1867-09111867-09382019-07-0171710.5073/JfK.2019.07.02<em>In situ</em> immunofluorescence localization: A method for rapid detection of <em>Beauveria</em> spp. in the rhizosphere of <em>Quercus robur</em> saplingsMarta Matek0Cornelia I. Ullrich1Frank Rabenstein2Eckhard Koch3Regina G. Kleespies4Croatian Forest Research Institute, Division for Forest Protection and Game Management, Jastrebarsko, CroatiaJulius Kühn-Institut (JKI), Federal Research Centre for Cultivated Plants, Institute for Biological Control, Darmstadt, GermanyJulius Kühn-Institut (JKI), Federal Research Centre for Cultivated Plants, Institute for Epidemiology and Pathogen Diagnostics, Quedlinburg, GermanyJulius Kühn-Institut (JKI), Federal Research Centre for Cultivated Plants, Institute for Biological Control, Darmstadt, GermanyJulius Kühn-Institut (JKI), Federal Research Centre for Cultivated Plants, Institute for Biological Control, Darmstadt, GermanyFor biological control of plant pests, e.g. cockchafer grubs, in the rhizosphere of oak, apple or pine trees, entomopathogenic Beauveria spp. are increasingly applied. For successful use, it is important to monitor the spread and persistence of the inoculated fungi, both qualitatively and quantitatively. The determination of both parameters by plating on selective nutrient media or by molecular methods such as PCR of soil samples are quite laborious and often do not yield satisfactory results. Therefore, the aim of the present study was to develop a specific in situ method using immunofluorescence labelling of Beauveria spp. growing on young fine roots of three-year old oak saplings. All fine roots investigated were covered with a dense net of soil rhizosphere fungi, as visualized by staining with the nonspecific dye blankophor. On non-inoculated roots, polyclonal Beauveria antibodies did not label any of these naturally growing fungi. Only samples of roots inoculated with Beauveria brongniartii displayed specific labelling up to ten months after inoculation. Whereas the natural rhizosphere fungi were detected growing in the intercellular space of the root cortex in an ectomycorrhiza-like manner up to the endodermis, hyphae of the inoculated B. brongniartii were never seen within the root tissue but only growing on the surface of the rhizodermis. These observations indicate that B. brongniartii does not grow endophytically, and that the method used allows to discriminate B. brongniartii from the resident fungal flora in the oak tree rhizosphere. Detection by immunofluorescence labelling employed in the current study may be a useful tool to follow B. brongniartii in experiments aimed at establishing the entomopathogen in the rhizosphere and to monitor its fate in long-term control of entomopathogens.https://ojs.openagrar.de/index.php/Kulturpflanzenjournal/article/view/13009biological pest controlblastosporesentomopathogenic fungiimmuno-fluorescence microscopyQuercus robursustainability. |
| spellingShingle | Marta Matek Cornelia I. Ullrich Frank Rabenstein Eckhard Koch Regina G. Kleespies <em>In situ</em> immunofluorescence localization: A method for rapid detection of <em>Beauveria</em> spp. in the rhizosphere of <em>Quercus robur</em> saplings Journal für Kulturpflanzen biological pest control blastospores entomopathogenic fungi immuno-fluorescence microscopy Quercus robur sustainability. |
| title | <em>In situ</em> immunofluorescence localization: A method for rapid detection of <em>Beauveria</em> spp. in the rhizosphere of <em>Quercus robur</em> saplings |
| title_full | <em>In situ</em> immunofluorescence localization: A method for rapid detection of <em>Beauveria</em> spp. in the rhizosphere of <em>Quercus robur</em> saplings |
| title_fullStr | <em>In situ</em> immunofluorescence localization: A method for rapid detection of <em>Beauveria</em> spp. in the rhizosphere of <em>Quercus robur</em> saplings |
| title_full_unstemmed | <em>In situ</em> immunofluorescence localization: A method for rapid detection of <em>Beauveria</em> spp. in the rhizosphere of <em>Quercus robur</em> saplings |
| title_short | <em>In situ</em> immunofluorescence localization: A method for rapid detection of <em>Beauveria</em> spp. in the rhizosphere of <em>Quercus robur</em> saplings |
| title_sort | em in situ em immunofluorescence localization a method for rapid detection of em beauveria em spp in the rhizosphere of em quercus robur em saplings |
| topic | biological pest control blastospores entomopathogenic fungi immuno-fluorescence microscopy Quercus robur sustainability. |
| url | https://ojs.openagrar.de/index.php/Kulturpflanzenjournal/article/view/13009 |
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