Improvements in Protoplast Isolation Protocol and Regeneration of Different Cabbage (<i>Brassica oleracea</i> var. <i>capitata</i> L.) Cultivars
Protoplasts are a versatile tool in plant biotechnology since they can be used for basic biological studies as well as for breeding strategies based on genome editing. An efficient protoplast isolation protocol is essential for conducting protoplast-based studies. To optimize the protoplast isolatio...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-08-01
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| Series: | Plants |
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| Online Access: | https://www.mdpi.com/2223-7747/12/17/3074 |
| _version_ | 1797581969472618496 |
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| author | Ester Stajič |
| author_facet | Ester Stajič |
| author_sort | Ester Stajič |
| collection | DOAJ |
| description | Protoplasts are a versatile tool in plant biotechnology since they can be used for basic biological studies as well as for breeding strategies based on genome editing. An efficient protoplast isolation protocol is essential for conducting protoplast-based studies. To optimize the protoplast isolation protocol in cabbage (<i>Brassica oleracea</i> var. <i>capitata</i> L.), different enzyme solutions were tested for the isolation of leaf mesophyll protoplasts. In our experiments, the combination of 0.5% Cellulase Onozuka RS and 0.1% Macerozyme R-10 showed the best result. The optimized protocol proved suitable for the isolation of protoplasts from five different cabbage cultivars with yields ranging from 2.38 to 4.63 × 10<sup>6</sup> protoplasts/g fresh weight (fw) and a viability of 93% or more. After three weeks in culture, protoplasts from all of the tested cultivars formed micro-calli, but further callus growth and shoot regeneration depended strongly on the genotype and regeneration protocol used. For shoot formation, 1 mg/L BAP in combination with auxin 0.2 mg/L NAA showed the best results with a regeneration of 23.5%. The results obtained will contribute to the development of different applications of cabbage protoplasts and facilitate the breeding process of this important horticultural crop. |
| first_indexed | 2024-03-10T23:15:14Z |
| format | Article |
| id | doaj.art-3abc619bdfa64f34beaff8047554989a |
| institution | Directory Open Access Journal |
| issn | 2223-7747 |
| language | English |
| last_indexed | 2024-03-10T23:15:14Z |
| publishDate | 2023-08-01 |
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| record_format | Article |
| series | Plants |
| spelling | doaj.art-3abc619bdfa64f34beaff8047554989a2023-11-19T08:40:57ZengMDPI AGPlants2223-77472023-08-011217307410.3390/plants12173074Improvements in Protoplast Isolation Protocol and Regeneration of Different Cabbage (<i>Brassica oleracea</i> var. <i>capitata</i> L.) CultivarsEster Stajič0Department of Agronomy, Biotechnical Faculty, University of Ljubljana, 1000 Ljubljana, SloveniaProtoplasts are a versatile tool in plant biotechnology since they can be used for basic biological studies as well as for breeding strategies based on genome editing. An efficient protoplast isolation protocol is essential for conducting protoplast-based studies. To optimize the protoplast isolation protocol in cabbage (<i>Brassica oleracea</i> var. <i>capitata</i> L.), different enzyme solutions were tested for the isolation of leaf mesophyll protoplasts. In our experiments, the combination of 0.5% Cellulase Onozuka RS and 0.1% Macerozyme R-10 showed the best result. The optimized protocol proved suitable for the isolation of protoplasts from five different cabbage cultivars with yields ranging from 2.38 to 4.63 × 10<sup>6</sup> protoplasts/g fresh weight (fw) and a viability of 93% or more. After three weeks in culture, protoplasts from all of the tested cultivars formed micro-calli, but further callus growth and shoot regeneration depended strongly on the genotype and regeneration protocol used. For shoot formation, 1 mg/L BAP in combination with auxin 0.2 mg/L NAA showed the best results with a regeneration of 23.5%. The results obtained will contribute to the development of different applications of cabbage protoplasts and facilitate the breeding process of this important horticultural crop.https://www.mdpi.com/2223-7747/12/17/3074<i>Brassica oleracea</i>protoplastsisolationregeneration |
| spellingShingle | Ester Stajič Improvements in Protoplast Isolation Protocol and Regeneration of Different Cabbage (<i>Brassica oleracea</i> var. <i>capitata</i> L.) Cultivars Plants <i>Brassica oleracea</i> protoplasts isolation regeneration |
| title | Improvements in Protoplast Isolation Protocol and Regeneration of Different Cabbage (<i>Brassica oleracea</i> var. <i>capitata</i> L.) Cultivars |
| title_full | Improvements in Protoplast Isolation Protocol and Regeneration of Different Cabbage (<i>Brassica oleracea</i> var. <i>capitata</i> L.) Cultivars |
| title_fullStr | Improvements in Protoplast Isolation Protocol and Regeneration of Different Cabbage (<i>Brassica oleracea</i> var. <i>capitata</i> L.) Cultivars |
| title_full_unstemmed | Improvements in Protoplast Isolation Protocol and Regeneration of Different Cabbage (<i>Brassica oleracea</i> var. <i>capitata</i> L.) Cultivars |
| title_short | Improvements in Protoplast Isolation Protocol and Regeneration of Different Cabbage (<i>Brassica oleracea</i> var. <i>capitata</i> L.) Cultivars |
| title_sort | improvements in protoplast isolation protocol and regeneration of different cabbage i brassica oleracea i var i capitata i l cultivars |
| topic | <i>Brassica oleracea</i> protoplasts isolation regeneration |
| url | https://www.mdpi.com/2223-7747/12/17/3074 |
| work_keys_str_mv | AT esterstajic improvementsinprotoplastisolationprotocolandregenerationofdifferentcabbageibrassicaoleraceaivaricapitatailcultivars |