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A More Accessible, Time-Saving, and Efficient Method for In Vitro Plant Regeneration from Potato Protoplasts
Both obtaining high-yielding, viable protoplasts and following reliable regeneration protocols are prerequisites for the continuous expansion and development of newly emerging systems involving protoplast utilization. This study determines an efficient process from protoplast isolation to shoot rege...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8071491/ https://www.ncbi.nlm.nih.gov/pubmed/33923378 http://dx.doi.org/10.3390/plants10040781 |
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author | Moon, Ki-Beom Park, Ji-Sun Park, Su-Jin Lee, Hyo-Jun Cho, Hye-Sun Min, Sung-Ran Park, Youn-Il Jeon, Jae-Heung Kim, Hyun-Soon |
author_facet | Moon, Ki-Beom Park, Ji-Sun Park, Su-Jin Lee, Hyo-Jun Cho, Hye-Sun Min, Sung-Ran Park, Youn-Il Jeon, Jae-Heung Kim, Hyun-Soon |
author_sort | Moon, Ki-Beom |
collection | PubMed |
description | Both obtaining high-yielding, viable protoplasts and following reliable regeneration protocols are prerequisites for the continuous expansion and development of newly emerging systems involving protoplast utilization. This study determines an efficient process from protoplast isolation to shoot regeneration in vitro. The maximum yield of protoplast extraction, which was 6.36 ± 0.51 × 10(6) protoplasts/g fresh weight (FW), was approximately 3.7 times higher than that previously reported for potato protoplasts. To obtain data, wounded leaves were used by partially cutting both sides of the midrib, and isolated protoplasts were purified by the sucrose cushion method, with a sucrose concentration of 20%. We confirmed a significant effect on the extraction efficiency by measuring enzymolysis during a 6 h period, with three times more washing buffer than the amount normally used. Protoplasts fixed in alginate lenses with appropriate space were successfully recovered and developed into microcalli 2 weeks after culture. In addition, to induce high efficiency regeneration from protoplasts, calli in which greening occurred for 6 weeks were induced to develop shoots in regeneration medium solidified by Gelrite, and they presented a high regeneration efficiency of 86.24 ± 11.76%. |
format | Online Article Text |
id | pubmed-8071491 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-80714912021-04-26 A More Accessible, Time-Saving, and Efficient Method for In Vitro Plant Regeneration from Potato Protoplasts Moon, Ki-Beom Park, Ji-Sun Park, Su-Jin Lee, Hyo-Jun Cho, Hye-Sun Min, Sung-Ran Park, Youn-Il Jeon, Jae-Heung Kim, Hyun-Soon Plants (Basel) Article Both obtaining high-yielding, viable protoplasts and following reliable regeneration protocols are prerequisites for the continuous expansion and development of newly emerging systems involving protoplast utilization. This study determines an efficient process from protoplast isolation to shoot regeneration in vitro. The maximum yield of protoplast extraction, which was 6.36 ± 0.51 × 10(6) protoplasts/g fresh weight (FW), was approximately 3.7 times higher than that previously reported for potato protoplasts. To obtain data, wounded leaves were used by partially cutting both sides of the midrib, and isolated protoplasts were purified by the sucrose cushion method, with a sucrose concentration of 20%. We confirmed a significant effect on the extraction efficiency by measuring enzymolysis during a 6 h period, with three times more washing buffer than the amount normally used. Protoplasts fixed in alginate lenses with appropriate space were successfully recovered and developed into microcalli 2 weeks after culture. In addition, to induce high efficiency regeneration from protoplasts, calli in which greening occurred for 6 weeks were induced to develop shoots in regeneration medium solidified by Gelrite, and they presented a high regeneration efficiency of 86.24 ± 11.76%. MDPI 2021-04-16 /pmc/articles/PMC8071491/ /pubmed/33923378 http://dx.doi.org/10.3390/plants10040781 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Moon, Ki-Beom Park, Ji-Sun Park, Su-Jin Lee, Hyo-Jun Cho, Hye-Sun Min, Sung-Ran Park, Youn-Il Jeon, Jae-Heung Kim, Hyun-Soon A More Accessible, Time-Saving, and Efficient Method for In Vitro Plant Regeneration from Potato Protoplasts |
title | A More Accessible, Time-Saving, and Efficient Method for In Vitro Plant Regeneration from Potato Protoplasts |
title_full | A More Accessible, Time-Saving, and Efficient Method for In Vitro Plant Regeneration from Potato Protoplasts |
title_fullStr | A More Accessible, Time-Saving, and Efficient Method for In Vitro Plant Regeneration from Potato Protoplasts |
title_full_unstemmed | A More Accessible, Time-Saving, and Efficient Method for In Vitro Plant Regeneration from Potato Protoplasts |
title_short | A More Accessible, Time-Saving, and Efficient Method for In Vitro Plant Regeneration from Potato Protoplasts |
title_sort | more accessible, time-saving, and efficient method for in vitro plant regeneration from potato protoplasts |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8071491/ https://www.ncbi.nlm.nih.gov/pubmed/33923378 http://dx.doi.org/10.3390/plants10040781 |
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