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Low light and low ammonium are key factors for guayule leaf tissue shoot organogenesis and transformation

  • Genetic transformation and hybridization
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Abstract

A new method has been developed for guayule tissue culture and transformation. Guayule leaf explants have a poor survival rate when placed on normal MS medium and under normal culture room light conditions. Low light and low ammonium treatment greatly improved shoot organogenesis and transformation from leaf tissues. Using this method, a 35S promoter driven BAR gene and an ubiquitin-3 promoter driven GUS gene (with intron) have been successfully introduced into guayule. These transgenic guayule plants were resistant to the herbicide ammonium-glufosinate and were positive to GUS staining. Molecular analysis showed the expected band and signal in all GUS positive transformants. The transformation efficiency with glufosinate selection ranged from 3 to 6%. Transformation with a pBIN19-based plasmid containing a NPTII gene and then selection with kanamycin also works well using this method. The ratio of kanamycin-resistant calli to total starting explants reached 50% in some experiments.

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Acknowledgement

We thank William Crosby for providing pBI426 that contained the double 35S promoter and alfalfa mosaic virus enhancer (Datla et al. 1991); Dr William Belknap for providing the construct that contained the Ubiquitin-3 promoter and the first ubiquitin coding sequence (Garbarino and Belknap 1994); Dr Peter Quail for providing the BAR gene (Christensen and Quail 1996); Dr Pal Maliga for providing pPZP200 (Hajdukiewicz et al. 1994); Dr Elizabeth Hood for providing Agrobacterium strain EHA101; Dr Willmitzer for providing the intron containing GUS gene (Vancanneyt el al. 1990). We also thank Dr. James Thomson and Dr. Grace Chen for their critical reviews of this manuscript

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  1. Katrina Cornish

    Present address: Yulex Corporation, 1945 Camino Vida Roble, Suite C, Carlsbad, CA, 92008, USA

Authors and Affiliations

  1. USDA/ARS/WRRC/CIU, 800 Buchanan St., Albany, CA, 94710, USA

    Niu Dong & Katrina Cornish

  2. Mendel Biotechnology Inc., 21375 Cabot Blvd., Hayward, CA, 94545, USA

    Belen Montanez & Robert A. Creelman

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  1. Niu Dong
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  2. Belen Montanez
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Correspondence to Katrina Cornish.

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Communicated by G. C. Phillips

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Dong, N., Montanez, B., Creelman, R.A. et al. Low light and low ammonium are key factors for guayule leaf tissue shoot organogenesis and transformation. Plant Cell Rep 25, 26–34 (2006). https://doi.org/10.1007/s00299-005-0024-2

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