Optimization of Microprojectile Bombardment Parameters for Pollen-Mediated Transformation in Gossypium hirsutum L.
Oct 8, 2018
Mature pollen-mediated transformation in cotton (Gossypium hirsutum L.) by microprojectile bombardment was established under gaseous medium in our laboratories. Bombardment parameters influencing the vitality, in vitro germinability and GUS transient expression of bombarded pollen were studied and optimized by the Gray System Analysis method. Optimum conditions for cotton are summarized as follows: the microprojectiles were 0.8-1.1 m diameter metal (tungsten or gold) granules; target distance between the stopping screen and the tissue was 5 cm for a JQ-700 gene delivery device, or 8 cm for a PDS-1000/He apparatus; and mature pollen was bombarded one time at a velocity of 450 m/s for the JQ-700 gene delivery device, or under 1100 psi shot pressure for the PDS-1000/He apparatus. Seeds were set after pollinating with the bombarded pollen.