In Vitro Peanut Culture: From Seed to Seed.

In vitro plants are widely used in biotechnology studies. In general, tissue culture requires special skills in order to obtain healthy and fertile axenic plants. Peanuts are considered recalcitrant, meaning they are difficult to regenerate under in vitro conditions. Thus, in vitro completion of the peanut plant cycle is the limiting factor in projects that require plant and seed recovery when applying molecular tools such as CRISPR. The protocols presented here allow for obtaining multiple shoots by micropropagation and inducing these shoots to produce flowers that successfully result in fertile pegs and pods, completing the life cycle under in vitro conditions. This is a significant step to perform studies that require axenic conditions in their entirety. These protocols outline the steps required to micropropagate Georgia 06G peanut plants under aseptic conditions, prepare culture medium, maintain shoot multiplication cultures, induce flowering and pegging, and maintain culture conditions to allow the development of pods. Published 2023. This article is a U.S. Government work and is in the public domain in the USA. Basic Protocol 1: Seed sterilization and germination Basic Protocol 2: Shoot multiplication Basic Protocol 3: Flower induction Basic Protocol 4: Peg formation.