Spore Mutagenesis with X-Rays

Have an old x-ray machine lying around. Use x-rays to effectively mutagenize spores for selection protocols

Materials

  • X-ray machine
  • Petri Dish, glass
  • Plastic sandwich bag
  • C-Fern spores
  • Materials and supplies for spore sterilization and culture inoculation 

Methods

SOAK SPORE Soak spores for 24 hours prior to sterilization and irradiation.

STERILIZE SPORES Sterilize using standard methods and place sterile spore suspension in a glass petri dish. Place dish inside a plastic sandwich bag.

IRRADIATE SPORES Irradiate spores with 25,000 Roentgens or about 555 Roentgens min-1 for 45 min. After irradiation sow spores as usual.

EXAMPLE RESULTS Under these conditions, spore germination at 10 days following sowing is about 80%. A mutant screen on culture media supplemented with 10-5 M flurodeoxyuridine gives a mutant frequency of 2 X 10-5.

Thomas Warne