Abstract
An efficient procedure was developed for in vitro callus induction, proliferation and regeneration of carnation cultivar (Dianthus caryophyllus L.) using leaf, nodal and inter-nodal explants on Murashige and Skoog’s medium (MS) supplemented with exogenous plant growth regulators. For morphogenic callus induction and proliferation from various explants, MS medium supplemented with 3.0 mg/l 2,4-D was highly efficient with 100% callus induction frequency from inter-nodal explants. Leaf explants showed quicker response than nodal and inter-modal explants, for callus initiation within 6 days of inoculation. Best grown callus was obtained from leaf explant. The leaf-derived callus was maintained up to several weeks, which indicated that 8-week incubation period was the most suitable for obtaining well proliferated, morphogenic callus. Temperature variation also affected the growth of in vitro induced morphogenic callus from various explants. Results have shown that 27°C proved to be the best temperature for morphogenic callus induction and proliferation from leaf and inter-nodal explants. Among the auxin-cytokinin combination, MS medium containing 1.0 mg/l N(6)-benzylaminopurin (BAP) and 2.0 mg/l NAA showed the highest efficiency of callus initiation and proliferation from leaf, nodal and inter-nodal explants. Light conditions proved better for callogenesis and proliferation from leaf, nodal and inter-nodal explants. Regeneration response from well grown morphogenic callus was prominent on MS medium supplemented with 3.0 mg/l BAP alone and 1.0 mg/l NAA with 3.0 mg/l BAP.
Highlights
Carnation, a member of the family Caryophyllous, has 88 genera and 1750 species, which were cultivated over 2000 years ago in Asia and Europe
Efficient systems have been reported for the regeneration of plants from leaf base [5], stem explants [6] and for the induction of direct somatic embryogenesis from leaf, petal as well as anther explants by Pareek and Kothari [7], Karami et al [8], and Xiaopeng et al [9], but still more efficient protocols are needed to increase the number of plants in a shorter period of time
In present study callus induction was investigated using MS medium supplemented with different concentrations of 2,4-D ranging from 1 - 5.0 mg/l. leaf explants showed 100 % callus induction response within 6 days of inoculation on MS medium containing 3.0 mg/l 2,4-D (Table 1)
Summary
A member of the family Caryophyllous, has 88 genera and 1750 species, which were cultivated over 2000 years ago in Asia and Europe. Callogenesis has been used as a tool to study plant physiology, cell biology and genetics. It has been exploited in agricultural practices, forestry and industry in order to achieve mass-propagation of disease free plants. Plants regenerated via callogenesis exhibit heritable mutations which influence the plant morphology, flower color and shape. It is novel source of genetic variation which contradicts the concept of clonal uniformatiy in micropropagation. The main aim was to determine the optimum concentration of plant growth regulators, best source of explants and well suited micro environment conditions for callus induction, proliferation and regeneration of carnation cultivar (Dianthus caryophyllous L.)
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