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Conditions for biolistic-mediated transformation of Lilium longiflorum 'Nellie White'

Author:
Kamo, Kathryn
Source:
Floriculture and Ornamental Biotechnology 2013 v.7 no.1 pp. 71
ISSN:
1749-0294
Subject:
Lilium longiflorum, biolistics, bulbs, callus, cell suspension culture, dicamba, gene fusion, genetic transformation, gold, growing media, micropropagation, picloram, somatic embryogenesis, somatic embryos, transgenic plants
Abstract:
A variety of tissues were used for biolistic-mediated transformation of Lilium longiflorum "Nellie White". Transgenic plants were not recovered from five-month-old, "non-select" callus or suspension cells that had been bombarded with pDM327 that contains the bar-uidA fusion gene under control of the CaMV 35S promoter. In comparison, ten transgenic plants were recovered from "select" callus that had been selected for its embryogenic-like appearance. Transgenic plants were also obtained following direct bombardment of precultured bulb scales. Both preculture time and preculture medium had an impact on transformation frequencies. Bulb scales cultured overnight on Murashige and Skoog's medium containing 1.0 mg/L picloram before bombardment with 0.6 um gold resulted in a transformation frequency of 0.9%. A one week or one month preculture time on Murashige and Skoog's medium containing 1.0 mg/L picloram yielded higher, 4.2% and 5%, transformation frequencies, respectively. When bulb scales were cultured on Murashige and Skoog's medium with either 2.0 mg/L dicamba or 0.5 mg/L picloram, lower frequencies of transformation, 1.2% and 0.8%, respectively, were observed. Direct bombardment of bulb scales eliminates the time needed for callus induction and multiplication when embryogenic callus is bombarded.
Agid:
56978
Handle:
10113/56978