10 seeds of control versus 10 seeds of drug treated. From same batch of seeds. Chosen randomly assigned into each group to have identical matched groups.
Protocol:
(1) Incubate overnight at room temp in 1ml 15ml falcon tubes either distilled water or 5mg/ml pen-strep (stock concentration).
(2) Next day add 9ml of distilled water to the tubes and pour the seeds into standard dime baggies with perlite (so drug is now 0.5mg/ml). Pour off excess liquid as standard. Keep it at 4oC for re-use.
(3) Incubate at room temp for germination. At day 7 most had germinated. 10 for the control and 8 for the drug treated.
(4) Seeds photographed on blue post-its, and placed into small pots (groups kept together). Soil was pre-wetted with distilled water OR 0.5mg/ml (in distilled water) using a dropper. Excess was caught in small red plastic shot-glasses.
(5) Soil was maintained damp in the way above in 4 for the next week. Seedlings were left together under grow lights and photographed every day (sometimes twice per day, 12h apart - evening photography gives better resolution of colors).
(6) At day 14 albino or potential variegated were extracted and grown separately with the withdrawl of drugs, into fresh soil wetted with distilled water. Drug-treated green ones which looked normal were continued to be grown in same concentration of drug.
(7) At week 3 one of the 4 remaining drug-treated ones had a whitish-borders leaf emerging (one of the pics above). A few days later, however, it appeared more green. This was exposed to UV by moving it to a window before I saw the white, then moved back to the original place to more closely observe. Perhaps UV stimulation is required?
(8) Also note - preceeding (1) the seeds were first "mistakenly" incubated in the drug at 4oC for an hour or 2, before I read the protocol saying room temperature. This small detail may or may not be inconsequential, but is noted just in case.
