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United States Department of Agriculture

Agricultural Research Service

Research Project: GENETIC ENGINEERING OF FLORAL BULB CROPS FOR VIRUS AND NEMATODE RESISTANCE Title: Optimized conditions for biolistic-mediated transformation of Lilium longilforum 'Nellie White'

Author
item Kamo, Kathryn

Submitted to: Floriculture, Ornamental and Plant Biotechnology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: October 4, 2012
Publication Date: December 12, 2012
Citation: Kamo, K.K. 2012. Optimized conditions for biolistic-mediated transformation of Lilium longilforum 'Nellie White'. Floriculture, Ornamental and Plant Biotechnology. 7:71-76.

Interpretive Summary: Lilies are an important cut flower and popular as a landscape and pot plant. Genetic engineering offers the opportunity to develop lilies with resistance to the viral and fungal pathogens and nematodes that infect them. The main obstacle to engineering lilies is that the transformation efficiency has been low. In this study, conditions were optimized for the transformation of lily using the gene gun. It was found that bulb scales of lilies could be directly bombarded and transgenic plants obtained from them. The transformation efficiency using bulb scales was 5% which is the highest reported to date using this method of transformation on lilies. This will allow us to now genetically enhance lilies with various genes for resistance.

Technical Abstract: A variety of tissues were used for biolistic-mediated transformation of Lilum longiflorum 'Nellie White'. Transgenic plants were not recovered from five-month-old, non-embryogenic callus or suspension cells that had been bombarded with pDM327 that contains the bar-uidA fusion gene under control the CaMV 35S promoter. In comparison, ten transgenic plants were recovered from "select" callus that had been selected for it 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 cales 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 elimates the time needed for callus induction and multiplication when embryogenic callus is bombarded.

Last Modified: 11/23/2014
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