A more accessible, time-saving, and efficient method for in vitro plant regeneration from potato protoplasts

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Title
A more accessible, time-saving, and efficient method for in vitro plant regeneration from potato protoplasts
Author(s)
Ki Beom Moon; Ji-sun Park; Su-Jin Park; Hyo Jun LeeHye Sun ChoSung Ran Min; Y I Park; Jae Heung JeonHyun-Soon Kim
Bibliographic Citation
Plants-Basel, vol. 10, no. 4, pp. 781-781
Publication Year
2021
Abstract
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 × 106 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%.
Keyword
Solanum tuberosumIsolationPurificationAlginate lensCallus greening
ISSN
2223-7747
Publisher
MDPI
DOI
http://dx.doi.org/10.3390/plants10040781
Type
Article
Appears in Collections:
Division of Biomaterials Research > Plant Systems Engineering Research > 1. Journal Articles
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