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

Stajič, Ester (avtor)

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.

• Jezik: Angleški jezik
• Ključne besede: Brassica oleracea, protoplasts, isolation, regeneration, protocol, cabbage, cultivars
• Vrsta gradiva: Članek v reviji
• Tipologija: 1.01 - Izvirni znanstveni članek
• Organizacija: BF - Biotehniška fakulteta
• Status publikacije: Objavljeno
• Različica publikacije: Objavljena publikacija
• Leto izida: 2023
• Št. strani: 12 str.
• Številčenje: Vol. 12, iss. 17, art. 3074
• PID: 20.500.12556/RUL-151618
• UDK: 577.2
• ISSN pri članku: 2223-7747
• COBISS.SI-ID: 168164355
• Datum objave v RUL: 12.10.2023

Gradivo je del revije

• Naslov: Plants
• Skrajšan naslov: Plants
• Založnik: MDPI
• ISSN: 2223-7747
• COBISS.SI-ID: 523345433

Licence

• Licenca: CC BY 4.0, Creative Commons Priznanje avtorstva 4.0 Mednarodna
• Povezava: http://creativecommons.org/licenses/by/4.0/deed.sl
• Opis: To je standardna licenca Creative Commons, ki daje uporabnikom največ možnosti za nadaljnjo uporabo dela, pri čemer morajo navesti avtorja.

Sekundarni jezik

• Jezik: Slovenski jezik
• Ključne besede: zelje, izolacija protoplastov, protokol, regeneracija, izolacija

Projekti

• Financer: ARRS - Agencija za raziskovalno dejavnost Republike Slovenije
• Številka projekta: Z4-3215-2021
• Naslov: Optimizacija protokola za CRISPR/Cas9 in tarčno preurejanje nukleotidov pri zelju

• Financer: ARRS - Agencija za raziskovalno dejavnost Republike Slovenije
• Številka projekta: P4-0077-2019
• Naslov: Kmetijske rastline - genetika in sodobne tehnologije