Articles
CHRYSANTHEMUM INDICUM PROTOPLAST CALLUS INDUCTION AND CULTURE
Article number
961_15
Pages
139 – 145
Language
English
Abstract
While Acta Horticulturae publishes the proceedings of Symposia or Congresses, i.e. papers presented at those meetings, the current article in its final format had been published in Acta Physiologiae Plantarum prior to the publication of the volume of Acta Horticulturae.
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Worldwide, Chrysanthemum indicum is the second most important ornamental crop, following rose.
The breeding history is largely unknown and due to the hexaploid nature conventional breeding is complicated.
Protoplast fusion might be an alternative to create new variation.
As in the past, chrysanthemum protoplasts were nearly always recalcitrant towards regeneration, our objective was to compare different culture techniques and their effect on a specific genotype.
Subsequently, further callus development on medium enriched with several phytohormone combinations was monitored.
Four methods to induce microcalli formation in a protoplast culture of C. i. Yoko Ono were compared, using a half strength MS medium enriched with Kao & Michayluk vitamins throughout.
Culture in liquid medium (17.6 microcalli/105 protoplasts) was preferable to culture in solid agarose beads, although the efficiency of the latter could be improved by layering them on glass beads (12.5 vs. 0.83 microcalli/105 protoplasts). Culture of protoplasts on moistened filter paper was unsuccessful.
In liquid media, microcalli were induced efficiently and easily after six weeks.
These effects may be explained by reduced toxicity due to cell breakdown and improved aeration.
Afterwards, microcalli were transferred to half strength MS enriched with auxins (IAA, 2,4-D or NAA) and cytokinins (TDZ, BA or KIN). On kinetin enriched medium, further growth (10.8-12.9% of all explants) was induced to a lesser extent than on TDZ or BA enriched medium (18.7-20.8 and 20.8-21.7%, respectively). As for auxins, IAA (19.2-24.2%) gave the best result.
The best growth (32.5%) was achieved by combining 0.5 mg L-1 IAA and 0.5 mg L-1 BA.
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Worldwide, Chrysanthemum indicum is the second most important ornamental crop, following rose.
The breeding history is largely unknown and due to the hexaploid nature conventional breeding is complicated.
Protoplast fusion might be an alternative to create new variation.
As in the past, chrysanthemum protoplasts were nearly always recalcitrant towards regeneration, our objective was to compare different culture techniques and their effect on a specific genotype.
Subsequently, further callus development on medium enriched with several phytohormone combinations was monitored.
Four methods to induce microcalli formation in a protoplast culture of C. i. Yoko Ono were compared, using a half strength MS medium enriched with Kao & Michayluk vitamins throughout.
Culture in liquid medium (17.6 microcalli/105 protoplasts) was preferable to culture in solid agarose beads, although the efficiency of the latter could be improved by layering them on glass beads (12.5 vs. 0.83 microcalli/105 protoplasts). Culture of protoplasts on moistened filter paper was unsuccessful.
In liquid media, microcalli were induced efficiently and easily after six weeks.
These effects may be explained by reduced toxicity due to cell breakdown and improved aeration.
Afterwards, microcalli were transferred to half strength MS enriched with auxins (IAA, 2,4-D or NAA) and cytokinins (TDZ, BA or KIN). On kinetin enriched medium, further growth (10.8-12.9% of all explants) was induced to a lesser extent than on TDZ or BA enriched medium (18.7-20.8 and 20.8-21.7%, respectively). As for auxins, IAA (19.2-24.2%) gave the best result.
The best growth (32.5%) was achieved by combining 0.5 mg L-1 IAA and 0.5 mg L-1 BA.
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Authors
T. Eeckhaut, J. Van Huylenbroeck
Keywords
in vitro, microcallus, phytohormones, recalcitrant, regeneration, somatic hybridization
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