Kaul V et al, Plant Cell, Tissue and Organ Culture 21: 21-30 (1990)
Summary
Adventitious shoots were regenerated from leaf and stem explants of eleven chrysanthemum cultivars. The optimum medium for both explant types contained Murashige & Skoog basal medium supplemented with 5μM 6-benzylaminopurine and 5μM e-napthaleneacetic acid. Generally, stem explants were superior to leaf explants. There were large cultivar differences in shoot regeneration frequency with three cultivars failing to respond over a wide range of hormone combinations. Shoots on stem explants appeared mainly to originate from cortical cells which rapidly divided and ruptured the epidermis. Regenerated shoots could be easily rooted, transferred to glasshouse conditions, and grown to flowering. All regenerated plants had the same morphological characteristics compared to plants derived from nodes.
Objectives
Adventitious shoots formation from leaf and stem explants
Materials
The cultivars used were: 'Eve Gray', 'Hurricane','Montrose Pink', 'Vulcan', 'Royal Purple', 'Lillian Hoek Red', 'Woking Scarlet', 'Blue Ridge', 'Red Cap', 'Red Fandango' and 'Dark Chip'. All were obtained as certified pathogen-free cultures from the Plant Research Institute (Victoria, Australia) except 'Hurricane', 'Montrose Pink' and 'Eve Gray' which were obtained from a local commercial laboratory.
Explant
nodal explants and shoot tips
Initial culture
Shoot cultures were maintained by sub-culturing nodal explants and shoot tips at 6-week intervals on MS medium with 0.8% (w/v) Difco bacto agar and incubated at 25°C under a 16h/8h photoperiod.
Shoot multiplication
The optimum medium for both explant types contained Murashige & Skoog basal medium supplemented with 5μM 6-BA and 5μM e-NAA.
Rooting
on hormone-free MS media
Acclimation
Plants were transferred to peat pots containing vermiculite:perlite (1:1 v/v) and kept under high humidity for 2 weeks prior to transfer to potting mix and standard glasshouse conditions.
Planting
Cultivation conditions
in greenhouse
Traints of regenerants
A range of morphological characteristics including leaf shape and size, growth habit and flowering were regularly observed.