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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 growth and plant regeneration for callus of Lilium longiflorum cv. Nellie White

Authors
item Kamo, Kathryn
item Han, Bong Hee -

Submitted to: Floriculture, Ornamental and Plant Biotechnology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: December 14, 2011
Publication Date: December 20, 2012
Citation: Kamo, K.K., Han, B. 2012. Optimized growth and plant regeneration for callus of Lilium longiflorum cv. Nellie White. Floriculture, Ornamental and Plant Biotechnology. 6:72-76.

Interpretive Summary: Regeneration of plants in vitro is a critical step in genetic engineering. The growth and regeneration of plants from lily callus was characterized in this study in an effort to determine which auxin results in the fastest callus growth and highest number of plants regenerated from the callus. The auxin picloram at 0.5 mg/L in the medium resulted in the most callus induced. Regeneration rates were comparable when callus had been grown on either picloram or dicamba. These results will be useful in determining optimum tissue culture conditions to enable genetic engineering of lily.

Technical Abstract: The rates of growth and regeneration were compared for compact callus, friable callus, and suspension cells of Lilium longiflorum cv. Nellie White to determine the optimal culture conditions. The fresh weight was higher for compact callus induced from bulb scales cultured on Murashige and Skoog’s medium supplemented with picloram (0.5, 1, or 2 mg/L) as compared to dicamba (2, 4, or 8 mg/L). The highest frequencies of embryogenic callus induction (60-90%) occurred from compact callus cultured on either picloram (0.5, 1, or 2 mg/L) or dicamba (2 mg/L). Compact callus cultured on picloram (0.5, 1, or 2 mg/L) or dicamba (2, 4, or 8 mg/L) grew slowly with a 1.2X increase in FW/month as compared to suspension cells grown in 0.5 mg/L picloram that increased 1.7X/month. Regeneration rates were similar (23-35 plantlets/gFW callus) for compact callus cultured on either dicamba (2 or 4 mg/L) or picloram (0.5 or 1 mg/L), but 3% of the plantlets regenerated from dicamba were phenotypically abnormal, and none were abnormal with picloram. Suspension cells showed a lower regeneration rate than compact callus with a maximum of only 12 plantlets regenerated from one g fresh weight suspensions cells grown in 0.5 mg/L picloram. A fast-growing, friable callus was induced and selected from compact callus cultured on Murashige and Skoog’s medium with 2 mg/L dicamba and 9% sucrose but not 3, 6, or 12% sucrose. Friable callus grew 5X faster than compact callus and formed numerous somatic embryo-like structures when cultured on Murashige and Skoog’s medium with 1% activated charcoal, but only a few embryo-like structures germinated to form plants with roots.

Last Modified: 12/18/2014
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