MS plates containing GA4/7 for seeds with low germination rate
In case of low germination rate of seeds of a specific line, we try to use of gibberellic acid (GA4/7) to enhance germination. For this purpose GA4/7 (2:1) is added to the MS plates that we use for sterile germination at a final concentration of 10 µM. The plates should be prepared without selection drug (sulfadiazine).
MS plates are prepared as described on the Sul selection scheme page, replacing the addition of sul solution after autoclaving and cooling by the addition of GA stock solution.
Sterilisation and plating of seeds on GA plates is performed as described on the Sul selection scheme page. Shortly: After sterilization, the seeds are transferred to the GA plates and kept at 4-8 °C for 4-5 days in the dark. The plates are then transferred to a light room / growth chamber (18-20 °C, long day conditions). Plantlets are transferred to soil after approximately 3 weeks and further grown under long day conditions.
GA stock solution
10 mM GA4/GA7 (2 : 1; M = 330 g/mol) in EtOH
(GA4/GA7 can be obtained as a ready to use powder. Stock: 0.165 g GA4/GA7 for 50 mL EtOH.)
Use 1 mL of GA stock solution for one litre MS plate medium.
Note 1: GA treatment may or may not help in germination. The responsiveness of the seeds to GA is dependent on their age. For seeds from our GABI-Kat collection, which are (in 2012) about 10 to 12 years old, we observed a success rate of about 20%.
Note 2: Plants grown after GA-stimulated germination do not show a regular growth phenotype. Nevertheless, they are usually able to produce seeds.