I have found one of the easiest methods to get Lilium into tissue culture is to use immature lily flower buds. By selecting an unopened flower bud that is about an inch (2.5mm) long it is easy to obtain sterile plant material. The bud is sterilized for 20 minutes in bleach (20% bleach/90% water) with agitation during the sterilization process. Mildly shaking the mixture every 4-5 minutes for a few seconds.
buds soaking in bleach solutionexcised ovary before slicing
ovary being excised from opened budovary slices
The bud is then removed in the laminar flow hood or where you do your tissue culture work. The bud tip and base is removed and the petals and tepals are sliced longitudinally and the pistil is removed. The stigma and style is removed from the ovary and the ovary is sliced like a cucumber. The sliced pieces are then lasted in culture vessels that have media. The media is composed of Media is composed of 50% strength MS salts, 30gm sugar/L, 2ml PPM/L, 1mg NAA/L, 1mg BA/L and 7gm carrageenan/L.
ovary slices in a25mm tube
The small pieces of ovary tissue cut into 2mm lengths will swell to about the size of the eraser at the end of a pencil in approximately 3 months. I find at this point it is advantageous to transfer the swollen bud sections to media without the NAA and BA hormones. The swollen bud should then start to differentiate into plantlets or callus material in about another two months. Once plantlets form they will continue to divide into multiple plants with additional replating.
