Increased alkaloid production from cell cultures of the California poppy through elicitation and in situ extraction with XAD-7

Abstract

Plants have the ability to synthesize compounds that help protect against insects, fungus, and other predators. These compounds are often valued as pharmaceuticals. Benzophenanthridine alkaloids (BPAs) are one family of compounds produced by the California poppy that are valued for their anti-viral, anti-cancer, anti-microbial, and anti-fungal properties. However, low yield limits the recovery of these compounds directly from plants. As a result, we are investigating their production using cell and tissue culture. This study investigates the addition of an elicitor coupled with in situ product extraction for increasing BPA production from cultures of the California poppy. Elicitors mimic pathogen attack; in this experiment, a purified extract derived from yeast cells served as the elicitor (10 mg/g of purified yeast extract PYE). Through In situ product extraction a hybrophobic adsorbent acts as an external storage site for BPAs; in this experiment, Amberlite XAD-7 served as the adsorbent (0.24 g of XAD-7/well containing 5 mL of cells). After 96 hours of treatment, PYE addition increased total alkaloid by 20-fold while the addition of both PYE and XAD-7 increased production by 65-fold over the control (no PYE or XAD-7). In cultures with XAD-7 addition, the majority of the alkaloid content (30.8 mg/g DW) was found adsorbed onto the resin, with considerably less BPAs found in cells themselves (7.0 mg/g DW). In all cases minimal BPA was found in the media. This study demonstrates PYE elicitation in combination with XAD-7 extraction can considerably increase alkaloid production in cultures of the California poppy.