Screening Methods for Isolation of Biocontrol Epiphytic Yeasts against <i>Penicillium digitatum</i> in Lemons
Worldwide, the green rot caused by <i>Penicillium digitatum</i> is one of the most aggressive postharvest diseases of lemons. Searching for sustainable alternatives to chemical fungicides, epiphytic yeasts as potential biocontrol agents were isolated from citrus fruits using a tailor-mad...
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MDPI AG
2021-02-01
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| Series: | Journal of Fungi |
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| Online Access: | https://www.mdpi.com/2309-608X/7/3/166 |
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| author | Martina María Pereyra Mariana Andrea Díaz Fabricio Fabián Soliz-Santander Anja Poehlein Friedhelm Meinhardt Rolf Daniel Julián Rafael Dib |
| author_facet | Martina María Pereyra Mariana Andrea Díaz Fabricio Fabián Soliz-Santander Anja Poehlein Friedhelm Meinhardt Rolf Daniel Julián Rafael Dib |
| author_sort | Martina María Pereyra |
| collection | DOAJ |
| description | Worldwide, the green rot caused by <i>Penicillium digitatum</i> is one of the most aggressive postharvest diseases of lemons. Searching for sustainable alternatives to chemical fungicides, epiphytic yeasts as potential biocontrol agents were isolated from citrus fruits using a tailor-made selective medium. For disclosing their antagonistic potential against <i>P. digitatum</i>, obtained isolates were subjected to direct screening methods, both in vitro and in vivo. In the course of the primary in vitro screening that comprised dual culture assays, 43 yeast strains displaying antagonistic activities against the pathogen were selected. Subsequently, such strains were subjected to an in vivo screening that consisted of a microscale test, allowing the selection of six yeast strains for further analysis. In the final screening using macroscale in vivo tests, three strains (AcL2, AgL21, and AgL2) displaying the highest efficiencies to control <i>P. digitatum</i> were identified. The protection efficiencies in lemons were 80 (AcL2), 76.7 (AgL21), and 75% (AgL2). Based on sequence analysis of the PCR amplified D1/D2 domains of the 26S rRNA genes, they were identified as representatives of the species <i>Clavispora lusitaniae</i>. Interestingly, the strains exhibited a broad action spectrum among citrus fruits as they were also able to combat the green mold disease in grapefruit and two orange varieties. The direct screening methods applied in this study favored the recovery of efficient candidates for application as biological control agents to combat fungal infestations of citrus fruits. |
| first_indexed | 2024-03-09T00:31:52Z |
| format | Article |
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| issn | 2309-608X |
| language | English |
| last_indexed | 2024-03-09T00:31:52Z |
| publishDate | 2021-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Journal of Fungi |
| spelling | doaj.art-d1c6ccf0250b407993737408f0687fc22023-12-11T18:27:50ZengMDPI AGJournal of Fungi2309-608X2021-02-017316610.3390/jof7030166Screening Methods for Isolation of Biocontrol Epiphytic Yeasts against <i>Penicillium digitatum</i> in LemonsMartina María Pereyra0Mariana Andrea Díaz1Fabricio Fabián Soliz-Santander2Anja Poehlein3Friedhelm Meinhardt4Rolf Daniel5Julián Rafael Dib6Planta Piloto de Procesos Industriales Microbiológicos (PROIMI)—Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Av. Belgrano y Pje. Caseros, 4000 Tucumán, ArgentinaPlanta Piloto de Procesos Industriales Microbiológicos (PROIMI)—Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Av. Belgrano y Pje. Caseros, 4000 Tucumán, ArgentinaPlanta Piloto de Procesos Industriales Microbiológicos (PROIMI)—Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Av. Belgrano y Pje. Caseros, 4000 Tucumán, ArgentinaGenomic and Applied Microbiology & Göttingen Genomics Laboratory, Institute of Microbiology and Genetics, Georg-August University of Göttingen, 37077 Göttingen, GermanyInstitut für Molekulare Mikrobiologie und Biotechnologie (IMMB), Westfälische Wilhelms Universität Münster, 48149 Münster, GermanyGenomic and Applied Microbiology & Göttingen Genomics Laboratory, Institute of Microbiology and Genetics, Georg-August University of Göttingen, 37077 Göttingen, GermanyPlanta Piloto de Procesos Industriales Microbiológicos (PROIMI)—Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Av. Belgrano y Pje. Caseros, 4000 Tucumán, ArgentinaWorldwide, the green rot caused by <i>Penicillium digitatum</i> is one of the most aggressive postharvest diseases of lemons. Searching for sustainable alternatives to chemical fungicides, epiphytic yeasts as potential biocontrol agents were isolated from citrus fruits using a tailor-made selective medium. For disclosing their antagonistic potential against <i>P. digitatum</i>, obtained isolates were subjected to direct screening methods, both in vitro and in vivo. In the course of the primary in vitro screening that comprised dual culture assays, 43 yeast strains displaying antagonistic activities against the pathogen were selected. Subsequently, such strains were subjected to an in vivo screening that consisted of a microscale test, allowing the selection of six yeast strains for further analysis. In the final screening using macroscale in vivo tests, three strains (AcL2, AgL21, and AgL2) displaying the highest efficiencies to control <i>P. digitatum</i> were identified. The protection efficiencies in lemons were 80 (AcL2), 76.7 (AgL21), and 75% (AgL2). Based on sequence analysis of the PCR amplified D1/D2 domains of the 26S rRNA genes, they were identified as representatives of the species <i>Clavispora lusitaniae</i>. Interestingly, the strains exhibited a broad action spectrum among citrus fruits as they were also able to combat the green mold disease in grapefruit and two orange varieties. The direct screening methods applied in this study favored the recovery of efficient candidates for application as biological control agents to combat fungal infestations of citrus fruits.https://www.mdpi.com/2309-608X/7/3/166epiphytic yeastsscreening methodsbiological control<i>Penicillium digitatum</i>citrus fruits |
| spellingShingle | Martina María Pereyra Mariana Andrea Díaz Fabricio Fabián Soliz-Santander Anja Poehlein Friedhelm Meinhardt Rolf Daniel Julián Rafael Dib Screening Methods for Isolation of Biocontrol Epiphytic Yeasts against <i>Penicillium digitatum</i> in Lemons Journal of Fungi epiphytic yeasts screening methods biological control <i>Penicillium digitatum</i> citrus fruits |
| title | Screening Methods for Isolation of Biocontrol Epiphytic Yeasts against <i>Penicillium digitatum</i> in Lemons |
| title_full | Screening Methods for Isolation of Biocontrol Epiphytic Yeasts against <i>Penicillium digitatum</i> in Lemons |
| title_fullStr | Screening Methods for Isolation of Biocontrol Epiphytic Yeasts against <i>Penicillium digitatum</i> in Lemons |
| title_full_unstemmed | Screening Methods for Isolation of Biocontrol Epiphytic Yeasts against <i>Penicillium digitatum</i> in Lemons |
| title_short | Screening Methods for Isolation of Biocontrol Epiphytic Yeasts against <i>Penicillium digitatum</i> in Lemons |
| title_sort | screening methods for isolation of biocontrol epiphytic yeasts against i penicillium digitatum i in lemons |
| topic | epiphytic yeasts screening methods biological control <i>Penicillium digitatum</i> citrus fruits |
| url | https://www.mdpi.com/2309-608X/7/3/166 |
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