Indirect somatic embryogenesis induction of Papaver degenii and influence of gelling agents and elicitors

Papaverdegenii (Urum. & Jav.) Kuzmanov is a Bulgarian glacial relict which is reported as part of Papaveralpinum subsp. alpinum by some studies. The species has restricted distribution and populations with low density and contains valuable alkaloids. Therefore, conservation measures are needed. Indirect somatic embryogenesis was induced for the first time for this species. The influence of gelling agents (Gelrite and agar) and elicitors (jasmonic acid and methyl jasmonate) on the regeneration effectiveness was studied. Phytochemical analysis on the obtained plants was conducted. It was ascertained that Murashige and Skoog medium supplemented with 4.53 mu M 2,4-dichlorophenoxyacetic acid + 2.27 mu M thidiazuron + 2.22 mu M benzylaminopurine was the most suitable for embryogenic callus induction. The biggest number of somatic embryos during the maturation stage was formed on medium B5 solidified with Gelrite. However, medium 2B5 hardened with agar had a better effect on their further development and germination. During the germination stage, a bigger number of somatic embryos formed on the media solidified with agar. Methyl jasmonate influence depended on the stage it was applied at. It inhibited the germination of the somatic embryos when it was applied at the maturation stage and stimulated the germination when it was added at the germination stage. Jasmonic acid slightly affected the development of somatic embryos regardless of the stage of their development. Methyl jasmonate increased the percentage of alkaloid content in the developed plants. The main alkaloid in the investigated plant samples was the alkaloid amurensine.

Automated summary

This study successfully induced indirect somatic embryogenesis in the Bulgarian glacial relict, Papaverdegenii, for the first time. By investigating different culture media and elicitors, the most suitable conditions for callus induction and somatic embryo development were determined. Phytochemical analysis of the plants was also conducted.