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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...

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Autores principales: 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
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
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%.
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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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