Summary
A somatic embryogenesis protocol was developed for Eschscholzia californica Chan. (California poppy) using embryogenic cell suspensions and optimized media conditions. Rapidly-growing, finely-dispersed embryogenic cell suspension cultures were established from embryogenic callus and maintained in B5 liquid media supplemented with 0.5 mg 1−1 (2.26 μM) 2,4-dichlorophenoxyacetic acid. Culture conditions were optimized by investigating the effect of basal media composition, gyratory shaker speed, various carbon sources, different cytokinins, and AgNO3 on the efficiency of somatic embryogenesis. After 40 d in culture, the somatic embryos that formed were counted and their overall growth expressed as pecked cell volume. The selected media consisted of either Gamborg (B5) or Murashige and Skoog (MS) salts and vitamins supplemented with 40 g 1−1 (117 mM) sucrose, 0.05 mg 1−1 (0.22 μM) 6-benzylaminopurine, and 10 mg l−1 (58.8 μM) AgNO3. Somatic embryo production was substantially reduced at shaker speeds above 40 rpm. Glucose and snerose were the most effective carbon sources, whereas fructose, galactose, and maltose resulted in a reduced yield and growth of somatic embryos. The development of somatic embryos was promoted by AgNO3 at concentrations below 10 mg l−1 (58.8 μM). A semi-solid medium containing 1.5 g l−1 Gel-rite produced the highest frequency of somatic embryo conversion, and promoted the efficient growth of plantlets. Using the reported protocol, over 500 viable somatic embryos were produced per 25 ml of embryogenic cell suspension culture.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Archambault, J.; Williams, R. D.; Lavoic, L.; Pepin, M. F.; Chavarie, C. Production of somatic embryos in helical-ribbon-impeller bioreactor. Biotechnol. Bioengineer. 44:930–943; 1994.
Gamborg, O. L.; Miller, R. O.; Ojima, K. Nutrient requirements of suspension cultures of soybean root cells. Exp. Cell Res. 50:151–158; 1968.
Gundlach, H.; Mueller, M. J.; Kutchan, T. M.; Zenk, M. H. Jasmonic acid is a signal transducer in elicitor-induced plant cell cultures. Proc. Natl Acad. Sci. USA 89:2389–2393; 1992.
Kavathekar, A. K.; Gamapathy, P. S. Embryoid differentiation in Eschscholzia californica. Curr. Sci. 42:671–673; 1973
Kavathekar, A. K.; Ganapathy, P. S.; Johri, B. M. Chilling induces development of embryoids into plantlets in Eschscholzia. Z. Pflanzenphysiol. 81:358–363; 1977.
Kutchan, T. M.. Molecular genetics of plant alkaloid biosynthesis. In: Cordell, G., ed The Alkaloids, Vol. 50. San Diego: Academic Press; 1998:257–316
Merkle, S. A.; Parrott, W. A.; Flinn, B. S.. Morphgenic aspects of somatic embryogenesis, In: Thorpe, T. A., ed In vitro embryogenesis in plants, Dordrecht: Kluwer; 1995:174–175.
Murashige, T.; Skoog, F. A revised medium for rapid growth and bioassays with tobacco tisue cultures. Physiol. Plant. 15:473–497; 1962.
Park, S. U.; Facchini, P. J. High-efficiency somatic embryogenesis and plant regeneration in California poppy. Eschscholzia californica Cham. Plant Cell Rep. 19:421–426; 2000
Purnhauser, L.; Medgyesy, P.; Czako, M.; Dix, P. J.; Marton, L. Stimulation of shoot regeneration in Triticum aestivum and Nicotiana plumbaginifolia Viv, tisue cultures using the ethylene inhibitor AgNO3. Plant Cell Rep. 6:1–4; 1987.
Reidiboym-Talleux, L; Diemer, F.; Sourdioux, M.; Chapelain, K.; Grenier-de March, G.. Improvement of soamtic embryogenesis in wild cherry (Prunus avium). Effect of maltose and ABA supplements. Plant Cell Tiss. Organ Cult. 55:199–209; 1999.
Roustan, J. P.; Latche, A.; Fallot, J. Control of carrot somatic embryogenesis by AgNo3, and inhibitor of ethylene action: effect on arginine decarboxylase activity. Plant Sci. 67:89–95; 1990.
Schumacher, H.-M.; Gundlach, H.; Fiedler, F.; Zenk, M. H. Elicitation of benzophenanthridine alkaloid synthesis in Eschscholtzia californica cell cultures. Plant Cell Rep 6:410–413; 1987.
Strickland, S. G.; Nichol, J. W.; McCall, C. M.; Stuart, D. A. Effect of carbohydrate source on alfalfa somatic embryogenesis. Plant Sci.. 48:113–121; 1987.
Tsay, H. S.; Huang, H. L. Somatic embryo formation and gemination from immature embryo-derived suspension-cultured cells of Angelica sinensis (Oliv.) Diels. Plant Cell Rep. 17:670–674; 1998.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Park, SU., Facchini, P.J. Somatic embryogenesis from embryogenic cell suspension cultures of california poppy, Eschscholzia californica Cham. In Vitro Cell.Dev.Biol.-Plant 37, 35–39 (2001). https://doi.org/10.1007/s11627-001-0007-0
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/s11627-001-0007-0