TY - JOUR

T1 - Protoplast isolation and culture from Kalanchoë species

T2 - optimization of plant growth regulator concentration for efficient callus production

AU - Cui, Jin

AU - Kuligowska Mackenzie, Kathryn

AU - Eeckhaut, Tom

AU - Müller, Renate

AU - Lütken, Henrik

PY - 2019/8/1

Y1 - 2019/8/1

N2 - A high yield of isolated protoplasts and efficient regeneration protocols are prerequisites for successful development of somatic hybrids. In the present study, protoplast isolation and regeneration were evaluated in 12 Kalanchoë accessions belonging to nine species. The highest protoplast yield was obtained from K. blossfeldiana ‘Charming Red Meadow’ with 10.78 ± 0.51 × 105 protoplasts per gram fresh weight. We observed significant differences of protoplast yield while there was no distinct difference in viability among the accessions. Seven accessions reached the microcolony stage and four developed microcalli in medium supplemented with 1.0 mg/l 1-naphthaleneacetic acid (NAA), 0.5 mg/l 6-benzylaminopurine (BAP) and 0.5 mg/l 2,4-dichlorophenoxy acetic acid (2,4-D). Using five selected accessions we optimized the PGR (plant growth regulators) concentrations using combinations of NAA, BAP and 2,4-D. K. blossfeldiana cultivars ‘Charming Red Meadow’ and ‘Paris’ produced significantly different numbers of calli depending on the PGR concentrations. For plant regeneration, the medium was supplemented with 1 mg/l NAA and 2 mg/l BAP or 2 mg/l zeatin. Shoots were regenerated on medium supplemented with NAA and BAP for K. blossfeldiana ‘Charming Red Meadow’ and K. blossfeldiana ‘Paris’. The plants successfully developed roots on the medium supplemented with IAA. The medium containing zeatin induced root formation directly from callus in K. blossfeldiana ‘Charming Red Meadow’. Our findings have the potential to facilitate the use of Kalanchoë species in somatic hybridization breeding programs.

AB - A high yield of isolated protoplasts and efficient regeneration protocols are prerequisites for successful development of somatic hybrids. In the present study, protoplast isolation and regeneration were evaluated in 12 Kalanchoë accessions belonging to nine species. The highest protoplast yield was obtained from K. blossfeldiana ‘Charming Red Meadow’ with 10.78 ± 0.51 × 105 protoplasts per gram fresh weight. We observed significant differences of protoplast yield while there was no distinct difference in viability among the accessions. Seven accessions reached the microcolony stage and four developed microcalli in medium supplemented with 1.0 mg/l 1-naphthaleneacetic acid (NAA), 0.5 mg/l 6-benzylaminopurine (BAP) and 0.5 mg/l 2,4-dichlorophenoxy acetic acid (2,4-D). Using five selected accessions we optimized the PGR (plant growth regulators) concentrations using combinations of NAA, BAP and 2,4-D. K. blossfeldiana cultivars ‘Charming Red Meadow’ and ‘Paris’ produced significantly different numbers of calli depending on the PGR concentrations. For plant regeneration, the medium was supplemented with 1 mg/l NAA and 2 mg/l BAP or 2 mg/l zeatin. Shoots were regenerated on medium supplemented with NAA and BAP for K. blossfeldiana ‘Charming Red Meadow’ and K. blossfeldiana ‘Paris’. The plants successfully developed roots on the medium supplemented with IAA. The medium containing zeatin induced root formation directly from callus in K. blossfeldiana ‘Charming Red Meadow’. Our findings have the potential to facilitate the use of Kalanchoë species in somatic hybridization breeding programs.

KW - Organogenesis

KW - Plant growth regulators

KW - Plant regeneration

KW - Protoplast isolation

KW - Protoplast-derived callus

UR - http://www.scopus.com/inward/record.url?scp=85066032802&partnerID=8YFLogxK

U2 - 10.1007/s11240-019-01624-4

DO - 10.1007/s11240-019-01624-4

M3 - Journal article

AN - SCOPUS:85066032802

VL - 138

SP - 287

EP - 297

JO - Plant Cell, Tissue and Organ Culture

JF - Plant Cell, Tissue and Organ Culture

SN - 0167-6857

IS - 2

ER -