High root biomass production in anchored Arabidopsis plants grown in axenic sucrose supplemented liquid culture

Abstract

There are many benefits to growing Arabidopsis in solution-based media, especially when large amounts of root tissue are required for molecular and biochemical studies. Roots grown in soil are brittle and tend to break easily when removed from their substrate. We have developed an axenic liquid culture system that simplifies growing large amounts of roots from intact plants. This technique consists of germinating 15 seeds on 2.5 cm2 stainless steel screens placed on half-strength semisolid Murashige and Skoog medium containing 1% or 2% sucrose. The screens anchor and support the plantlets in an upright position while keeping the roots and shoots separate. The seedlings are transferred with forceps to 125-mL wide-mouth Erlenmeyer flasks containing 10 mL of half-strength Murashige and Skoog liquid medium and 1% sucrose. The flasks are placed onto a floor rotary shaker under fluorescent lights. After 3 days, the sucrose is increased to 3% and the volume to 15 mLfor 7 days. During any further experimental manipulations, sucrose is not supplied. The media is changed every 3–4 days to replenish the nutrients. The presence of sucrose in the media dramatically increases the biomass, and large amounts of root tissue can easily be harvested.