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Title: Production of Pharmaceuticals from Papaver Cultivars In Vitro

item Tisserat, Brent
item Berhow, Mark

Submitted to: Engineering in Life Sciences
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 4/8/2009
Publication Date: 9/28/2009
Citation: Tisserat, B., Berhow, M.A. 2009. Production of Pharmaceuticals from Papaver Cultivars In Vitro. Engineering in Life Sciences. 9(3):190-196.

Interpretive Summary: This research determined that there is commercial possibility to produce pharmaceutical alkaloids (Benzyliosoquinoline alkaloids) within a controlled laboratory environment. Opium alkaloids are in great demand in the medical field since they are employed as important pain-killers. Iranian poppy (Papaver bracteatum Lindl.) and opium poppy (P. somniferum L.) are cool-season crops that provide these important medicinals; however, they can only be obtained through importation. As an alternative, we studied the possibility of producing pharmaceutical alkaloids within the laboratory environment from plant tissue cultures of opium and Iranian poppies. This was done by investigating the basic genetic, chemical and physical parameters associated with obtaining alkaloid-producing foliage in a sterile environment. The result indicates that continued development of such a system has the potential for commercial production of pharmaceutical alkaloids within a controlled laboratory environment, providing an alternative to importation.

Technical Abstract: A methodology to clonally proliferate Iranian poppy (Papaver bracteatum Lindl.) and opium poppy (P. somniferum L.) shoots is presented employing an in vitro hydroponics system (i.e., automated plant culture system (APCS)). Temperature had a profound effect on growth and alkaloid production after 8-weeks in culture. Poppy plantlets were obtained from germinated seeds on a Murashige and Skoog medium (MS). Plantlets of poppy cvs. grew best at 18.5 and 20 deg. C compared to 15 or 25 deg. C. For example, ‘Princess Princess’ opium poppy plantlets grown under 18.5 deg. C exhibited a 50% increase in fresh weight and a 191% increase in total alkaloids compared to shoots grown under 25 deg. C. A morphogenetic and pharmaceutical alkaloid survey study was conducted with 24 Iranian poppy cvs. and 21 opium poppy cvs. after 8-weeks in culture. Total alkaloids in cultured Iranian and opium poppy cvs. tested ranged from 0 to 6.55 mg.g-DW-1. The alkaloid content profiles in Iranian and opium poppies were 94.7% and 67.3% thebaine, 2.1% and 1.1% morphine and 3.9% and 28.4% codeine, respectively. Prolific axillary branching could be obtained from poppy cvs. by shoot reculture to BM with 1.0 mg l-1 N6-benzyladenine (BA) and 0.01 mg l-1 '-naphthalene acetic acid (NAA) for an additional 16 weeks. The influence of vessel size on the growth responses of established poppy shoot clumps was tested by subculture on a variety of culture vessels for 8 weeks. Culture vessels tested included culture tubes (55 mm3 capacity (cap.)), baby food jars (143 mm3 cap.), Magenta GA-7 containers (365 mm3 cap.), and polycarbonate jars (1890 mm3 cap.) employing an APCS. Culture vessel capacity had a significant positive correlation on shoot length, fresh weight, leaf number, and shoot number. Shoot length, fresh weight, leaf number, and shoot number grown in the polycarbonate jar with APCS exhibited increases of 1-, 21.5-, 7.8-, and 8.3-x fold, respectively, compared to that obtained from shoots grown in culture tubes. Higher culture growth rates that occurred in the larger vessel sizes were correlated with lower alkaloid treatment-1 (mg alkaloid g-DW-1). However, the overall total alkaloid vessel-1 ((mg alkaloid g-DW-1) x g culture DW) increased because of greater biomass production per vessel. Alkaloid content was found to remain stable for shoots grown over a six month evaluation period.