USE OF ION BEAMS FOR IN VITRO MUTATION BREEDING OF ORNAMENTAL PLANTS-APPLICATION IN EUSTOMA GRANDIFLORUM

Authors:	S. Ohki, Y. Sakai, K. Ooba, K. Chatani
Keywords:	⁴He²⁺ H⁺ beams, shoot regeneration, mutation, DNA marker, SSR, RAPD
Abstract: Leaf segments of Eustoma grandiflorum were irradiated with ⁴He²⁺ and H⁺ beams at different doses. Shoot differentiation was achieved by culturing on medium with 5 μM of 6-(,-dimethylalylamino)purine. Irradiation with ⁴He²⁺ at 10 Gy completely inhibited shoot formation. The number of shoots forming from leaf segments of cultivars ‘Mickey Soft Pink’ and ‘Mickey Rose’ decreased as the dose increased from 3 to 7 Gy. However, no such decrease was observed with cv. ‘White Palace’. With an H⁺ beam at 15 Gy, shoot differentiation was completely suppressed for ‘Mickey Soft Pink’. However, leaf segments of ‘White Palace’ irradiated at 15 Gy formed shoots. Shoots from irradiated leaf segments were successfully acclimatized and cultured in the greenhouse. Among 250 plants investigated, two plants that were regenerated from leaf segments irradiated with ⁴He²⁺ beam at 7 Gy showed a high branching habit. Using primers of simple sequence repeats, the polymorphism of microsatellite regions of the total DNA was analyzed. Polymorphism was observed in plants that did not show morphological mutations. It is possible to use this mutation system to obtain breeding materials for this species.
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