Micropropagation Protocol and Genetic Stability of the <i>Salix myrtilloides</i> Plants Cultivated In Vitro
<i>Salix myrtilloides</i> L. is a relict species, threatened with extinction in many European countries. To prevent the loss of the species, tissue culture was established to produce plant material for reintroduction in natural habitats. Micropropagation was chosen as a method to obtain...
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MDPI AG
2023-01-01
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| Series: | Biology |
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| author | Marzena Parzymies Magdalena Pogorzelec Katarzyna Głębocka Elwira Sliwinska |
| author_facet | Marzena Parzymies Magdalena Pogorzelec Katarzyna Głębocka Elwira Sliwinska |
| author_sort | Marzena Parzymies |
| collection | DOAJ |
| description | <i>Salix myrtilloides</i> L. is a relict species, threatened with extinction in many European countries. To prevent the loss of the species, tissue culture was established to produce plant material for reintroduction in natural habitats. Micropropagation was chosen as a method to obtain new plants. <i>S. myrtilloides</i> shoots were disinfected with NaOCl, AgNO<sub>3</sub>, or with a two-step disinfection with NaOCl, and then placed on MS medium supplemented with BA at 1 mg·dm<sup>−3</sup> and IBA at 0.1 mg·dm<sup>−3</sup>. Regenerated shoots were cultivated in presence of BA, KIN, and 2iP to select the treatment with the highest multiplication rate. The obtained plants were acclimatized to ex vitro conditions. Inter-simple sequence repeat (ISSR) and flow cytometric analyses were conducted on in vitro regenerated plants to check their genetic stability. The best disinfection results were obtained when explants were treated with 1.5% NaOCl for 20 min. The highest multiplication rate and good quality plants were noted in the control media, without growth regualtors and in presence of kinetin at 0.5 mg·dm<sup>−3</sup>. Flow cytometry and ISSR analyses confirmed genetic stability in plantlets, which indicated the possibility to use the in vitro obtained plants for reintroduction. |
| first_indexed | 2024-03-11T09:08:20Z |
| format | Article |
| id | doaj.art-bd98acde708c402c89860ae7f9044626 |
| institution | Directory Open Access Journal |
| issn | 2079-7737 |
| language | English |
| last_indexed | 2024-03-11T09:08:20Z |
| publishDate | 2023-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biology |
| spelling | doaj.art-bd98acde708c402c89860ae7f90446262023-11-16T19:12:32ZengMDPI AGBiology2079-77372023-01-0112216810.3390/biology12020168Micropropagation Protocol and Genetic Stability of the <i>Salix myrtilloides</i> Plants Cultivated In VitroMarzena Parzymies0Magdalena Pogorzelec1Katarzyna Głębocka2Elwira Sliwinska3Institute of Horticultural Production, University of Life Sciences in Lublin, 20-612 Lublin, PolandDepartment of Hydrobiology and Protection of Ecosystems, University of Life Sciences in Lublin, 20-262 Lublin, PolandInstitute of Plant Genetics, Breeding and Biotechnology, University of Life Sciences in Lublin, 20-950 Lublin, PolandLaboratory of Molecular Biology and Cytometry, Bydgoszcz University of Science and Technology, 85-796 Bydgoszcz, Poland<i>Salix myrtilloides</i> L. is a relict species, threatened with extinction in many European countries. To prevent the loss of the species, tissue culture was established to produce plant material for reintroduction in natural habitats. Micropropagation was chosen as a method to obtain new plants. <i>S. myrtilloides</i> shoots were disinfected with NaOCl, AgNO<sub>3</sub>, or with a two-step disinfection with NaOCl, and then placed on MS medium supplemented with BA at 1 mg·dm<sup>−3</sup> and IBA at 0.1 mg·dm<sup>−3</sup>. Regenerated shoots were cultivated in presence of BA, KIN, and 2iP to select the treatment with the highest multiplication rate. The obtained plants were acclimatized to ex vitro conditions. Inter-simple sequence repeat (ISSR) and flow cytometric analyses were conducted on in vitro regenerated plants to check their genetic stability. The best disinfection results were obtained when explants were treated with 1.5% NaOCl for 20 min. The highest multiplication rate and good quality plants were noted in the control media, without growth regualtors and in presence of kinetin at 0.5 mg·dm<sup>−3</sup>. Flow cytometry and ISSR analyses confirmed genetic stability in plantlets, which indicated the possibility to use the in vitro obtained plants for reintroduction.https://www.mdpi.com/2079-7737/12/2/168acclimatizationactive conservationcytokininsflow cytometryISSRmultiplication rate |
| spellingShingle | Marzena Parzymies Magdalena Pogorzelec Katarzyna Głębocka Elwira Sliwinska Micropropagation Protocol and Genetic Stability of the <i>Salix myrtilloides</i> Plants Cultivated In Vitro Biology acclimatization active conservation cytokinins flow cytometry ISSR multiplication rate |
| title | Micropropagation Protocol and Genetic Stability of the <i>Salix myrtilloides</i> Plants Cultivated In Vitro |
| title_full | Micropropagation Protocol and Genetic Stability of the <i>Salix myrtilloides</i> Plants Cultivated In Vitro |
| title_fullStr | Micropropagation Protocol and Genetic Stability of the <i>Salix myrtilloides</i> Plants Cultivated In Vitro |
| title_full_unstemmed | Micropropagation Protocol and Genetic Stability of the <i>Salix myrtilloides</i> Plants Cultivated In Vitro |
| title_short | Micropropagation Protocol and Genetic Stability of the <i>Salix myrtilloides</i> Plants Cultivated In Vitro |
| title_sort | micropropagation protocol and genetic stability of the i salix myrtilloides i plants cultivated in vitro |
| topic | acclimatization active conservation cytokinins flow cytometry ISSR multiplication rate |
| url | https://www.mdpi.com/2079-7737/12/2/168 |
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