Improvements in protoplast isolation protocol and regeneration of different cabbage (Brassica oleracea var. capitata L.) cultivars

Stajič, Ester (author)

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 (Brassica oleracea var. capitata 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 × 106 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.

• Language: English
• Keywords: Brassica oleracea, protoplasts, isolation, regeneration, protocol, cabbage, cultivars
• Work type: Article
• Typology: 1.01 - Original Scientific Article
• Organization: BF - Biotechnical Faculty
• Publication status: Published
• Publication version: Version of Record
• Year: 2023
• Number of pages: 12 str.
• Numbering: Vol. 12, iss. 17, art. 3074
• PID: 20.500.12556/RUL-151618
• UDC: 577.2
• ISSN on article: 2223-7747
• COBISS.SI-ID: 168164355
• Publication date in RUL: 12.10.2023

Record is a part of a journal

• Title: Plants
• Shortened title: Plants
• Publisher: MDPI
• ISSN: 2223-7747
• COBISS.SI-ID: 523345433

Licences

• License: CC BY 4.0, Creative Commons Attribution 4.0 International
• Link: http://creativecommons.org/licenses/by/4.0/
• Description: This is the standard Creative Commons license that gives others maximum freedom to do what they want with the work as long as they credit the author.

Secondary language

• Language: Slovenian
• Keywords: zelje, izolacija protoplastov, protokol, regeneracija, izolacija

Projects

• Funder: ARRS - Slovenian Research Agency
• Project number: Z4-3215-2021
• Name: Optimizacija protokola za CRISPR/Cas9 in tarčno preurejanje nukleotidov pri zelju

• Funder: ARRS - Slovenian Research Agency
• Project number: P4-0077-2019
• Name: Kmetijske rastline - genetika in sodobne tehnologije