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  • Title: Biolistic Transformation of Cotton Embryogenic Cell Suspension Cultures.
    Author: Rajasekaran K.
    Journal: Methods Mol Biol; 2019; 1902():55-66. PubMed ID: 30543061.
    Abstract:
    Genetic transformation of cotton (Gossypium hirsutum L.) is highly dependent on the ability to regenerate fertile plants from transgenic cells through somatic embryogenesis. Induction of embryogenic cell cultures is genotype-dependent. However, once embryogenic cell cultures are available, they can be effectively used for transformation by Agrobacterium or biolistic bombardment methods. Here I describe a detailed procedure to transform cotton embryogenic cell suspension cultures by biolistic bombardment. A commercially available, helium-driven biolistic device (Bio-Rad PDS1000/He) was used to bombard gold particles coated with plasmid DNA (for visual identification of transformed cells and/or selection) into embryogenic cells. Stable transformation at a high frequency (up to 4% of the transiently expressing cells) is possible. Regeneration of fertile transgenic plants from embryogenic cells takes only about 2 months. Another advantage of the embryogenic cell suspension cultures is that they are amenable for cryopreservation and long-term storage. It is highly preferable to transform commercial varieties of choice than obsolete varieties to avoid genetic drug due to backcrossing.
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