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ARS Home » Research » Publications at this Location » Publication #131563
Title: TRANSFORMATION OF SUGARBEET USING CELL SUSPENSION CULTURES

Author
item IVIC, SNEZANA - USDA ARS BELTSVILLE MD
item Saunders, Joseph
item Smigocki, Anna

Submitted to: Tissue Culture Association Congress Proceedings
Publication Type: Abstract Only
Publication Acceptance Date: 4/15/2002
Publication Date: N/A
Citation: N/A

Interpretive Summary:

Technical Abstract: A sugarbeet transformation method was developed using particle bombardment of short-term suspension cultures. Callus obtained from leaf discs of greenhouse-grown plants was propagated in liquid medium for 2 weeks and bombarded 1 day after sieving and plating onto agar medium. After 2 to 5 days callus was passed to medium containing either kanamycin or paramomycin and maintained on the selection media for different lengths of time. The introduced DNA contained both uidA gene (GUS) under the control of either the osmotin (OSM) or proteinase inhibitor II (Pin2) gene promoter and npt II gene under the control of the 35S promoter. Transient GUS expression monitored 2 days after bombardment showed 900 to 3000 blue units per bombarded plate of 0.2 g of suspension cells. GUS expression decreased significantly during the next 7-14 days of culture. Stably transformed calli were obtained as early as 3 weeks following bombardment at a frequency of 0.25 - 9 calli per bombarded plate. Presence of the introduced genes was confirmed by PCR analysis. To induce regeneration of plants, calli are being maintained on media with varying plant growth regulator composition.