333 Micropropagation of Echinacea angustifolia, E. pallida, and E. purpurea from Vegetative and Seed Explants
Micropropagation of three Echinacea species, E. angustifolia, E. pallida, and E. purpurea, was investigated as a potential means of germplasm preservation of species faced with overcollection in the wild and rapid clonal propagation of elite individuals with unique medicinal or ornamental properties. Comparison of explant sources indicated vegetative explants resulted in high contamination rates when collected from shoot-tips (100%),but not when collected from nodal explants (11% to 39). Seed coat removal reduced contamination from 100% in intact seeds to near 0% in excised embryos. Removal of seed coats (pericarp and integument layers) also eliminated dormancy requirement for germination. All species responded with shoot multiplication and loss of rooting when BA or thidiazuron was added to culture medium. Medium with thidiazuron resulted in excessive adventitious shoot formation. Shoot multiplication rates were low (one to three shoots/explant) on medium with BA levels low enough to avoid adventitious shoot formation. Medium containing half-strength MS minerals resulted in more shoots with smaller leaves than full-strength MS minerals. Cultures did not perform well on Woody Plant Medium. Increasing subculture frequency from every 4 weeks to every 2 weeks increased shoot multiplication rates from 1.4 to 1.8 shoots per subculture and total shoots produced after 12 weeks of culturing (per initial explant) from 2.8 to 23.9. Rooting occurred readily on shoots isolated from E. purpurea without addition of IBA. Rooting was low or non-existent on shoots from E. angustifolia and E. pallida, respectively, regardless of IBA level, light conditions, or temperature. Methods described in this study allow rapid multiplication of three Echinacea species and subsequent rooting of E. angustifolia and E. purpurea. Future improvements in root induction treatments will allow more effective use of micropropagation for Echinacea germplasm preservation and multiplication. Chemical names used: N-(phenylmethyl)-1H-purine-6-amine (BA), 1H-indole-3-butyric acid (IBA).