Protoplast Isolation and Shoot Regeneration from Protoplast-Derived Callus of Petunia hybrida Cv. Mirage Rose

Despite the increasing use of protoplasts in plant biotechnology research, shoot regeneration from protoplasts remains challenging. In this study, we investigated the factors involved in protoplast isolation, callus induction, and shoot regeneration in Petunia hybrida cv. Mirage Rose. The following...

Full description

Saved in:
Bibliographic Details
Published in:Biology (Basel, Switzerland) Switzerland), 2020-08, Vol.9 (8), p.228
Main Authors: Kang, Hyun Hee, Naing, Aung Htay, Kim, Chang Kil
Format: Article
Language:English
Subjects:
Biotechnology
Botanical research
Callus
Cell culture
Cell division
Cellulase
Chromosomes
CRISPR
Cultivars
digestion time
Enzymes
Gene expression
Genetic engineering
Genomes
Indole-3-butyric acid
Liquid culture
Mannitol
osmoticum conditions
Petunia
Petunia hybrida
Physiological aspects
plant growth regulators
Plant hormones
Plant tissue culture
plating density
protoplast viability
Protoplasts
Shoots
Shoots (Botany)
Sucrose
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Despite the increasing use of protoplasts in plant biotechnology research, shoot regeneration from protoplasts remains challenging. In this study, we investigated the factors involved in protoplast isolation, callus induction, and shoot regeneration in Petunia hybrida cv. Mirage Rose. The following conditions were found to be most optimal for protoplast yield and viability: 0.6 M mannitol, 2.0% cellulase, and 6 h digestion time. A plating density of 10 × 104 protoplasts/mL under osmoticum condition (0.58 M mannitol) showed high microcolony viability in liquid culture. The Kao and Michayluk medium was found to be appropriate for callus proliferation from microcalli under a 16-h light photoperiod. Calli cultured in Murashige and Skoog medium containing 1.0 mg/L 6-benzylaminopurine and 0.2 mg/L 3-indole butyric acid showed the highest shoot regeneration frequency and number of shoots obtained per explant. Random amplification of polymorphic DNA analysis showed that the protoplast-derived shoots exhibited the same banding patterns as those of donor plants. Collectively, these findings can contribute to solving problems encountered in protoplast isolation and shoot regeneration in other petunia cultivars and related species. As the protocol developed by us is highly reproducible, it can be applied in biotechnology research on P. hybrida cv. Mirage Rose.
ISSN:2079-7737
2079-7737
DOI:10.3390/biology9080228