Esterase and peroxidase isoforms during initial stages of somatic embryogenesis in Fritillaria meleagris L. leaf base

Marija Petrić, Angelina Subotić, Slađana Jevremović, Milana Trifunović-Momčilov, Vojin Tadić, Marica Grujić, Zoran Vujčić


The aim of this study was to determine the enzymatic profile of esterases and peroxidases during early stages of somatic embryogenesis of Fritillaria meleagris L. Somatic embryogenesis was induced using the leaf base as explant on a medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D). Zymography showed the presence of different moieties, six isoforms of esterases and peroxidases, during morphogenesis as compared to control explants. One isoform of esterases was detected only during the process of somatic embryogenesis, and one isoform was detected in control explants. Analysis of esterases with 1-naphthyl butyrate proved that esterases, which participate in somatic embryogenesis of F. meleagris, belong to the family of aryl esterases. For the first time it was proved that five isoforms of esterases, which are involved in morphogenesis of F. meleagris, belong to the family of aryl esterases, while two isoforms are carboxyl esterases. One isoform of carboxyl esterases was visible in control explants. This is also the first description of peroxidases during the morphogenetic process, and of the difference between aryl and carboxyl esterases. More isoforms of esterases during morphogenesis as compared to control explants are probably responsible for some early physiological process during somatic embryogenesis of F. meleagris.

Received: November 30, 2016; Revised: January 10, 2017; Accepted: March 3, 2017; Published online: March 10, 2017

How to cite this article: Petrić M, Subotić A, Jevremović S, Trifunović-Momčilov M, Tadić V, Grujić M, Vujčić Z. Esterase and peroxidase isoforms during initial stages of somatic embryogenesis in Fritillaria meleagris L. leaf base. Arch Biol Sci. 2017;69(4):619-25.


aryl esterase; esterase; F. meleagris; peroxidase; somatic embryogenesis

Full Text:



Li SL, Lin G, Chan SW, Li P. Determination of the major isosteroidal in bulbs of Fritillaria by high-performance liquid chromatography coupled with evaporative light scattering detection. J Chromatogr A. 2001;909:207-14.

Rahman A, Akhtar MN, Choudhary MI, Tsuda Y, Sener B, Khalid A, Parvez M. New steroidal alkaloids from Fritillaria imperialis and their cholinesterase inhibiting activities. Chem Pharm Bull. 2002;50:1013-7.

Sho I, Michiharu K, Testuo N. On the alkaloid of Fritillaria verticillate WILD. var. Thunbergii BAKER. II. The structure of verticine. Chem Pharm Bull. 1963;11:1337-40.

De Hertogh AA, Le Nard M. Physiological and biochemical aspects of flower bulbs. In: De Heroght AA, Le Nard M, editors. The physiology of flower bulbs. Amsterdam, The Netherlands: Elsevier; 1993. p. 53-69.

Fay M. Conservation of rare and endangered plants using in vitro methods. In vitro Cell Dev B. 1992;28:1-4.

Mirici S, Parmaksiz I, Ozcan S, Sancak C, Uranbey S, Sarihan EO, Gumuscu A, Gurbuz B, Arslan N. Efficient in vitro bulblet regeneration from immature embryos of endangered Sternbergia fischeriana. Plant Cell Tiss Org Cult. 2005;80:239-46.

Jevremović S, Petrić M, Živković S, Trifunović M, Subotić A. Superoxide dismutase activity and isoenzyme profiles in bulbs of Snake’s head fritillary in response to cold treatment. Arch Biol Sci. 2010;62:553-8.

Petrić M, Jevremović S, Trifunović M, Tadić V, Milošević S, Dragićević M, Subotić A. The effect of low temperature and GA3 treatments on dormancy breaking and activity of antioxidant enzymes in Fritillaria meleagris bulblets cultured in vitro. Acta Physiol Plant. 2013;35:3223-36.

Petrić M, Jevremović S, Trifunović M, Tadić V, Milošević S, Subotić A. Activity of antioxidant enzymes during induction of morphogenesis of Fritillaria meleagris in bulb scale culture. Turk J Biol. 2014;38:328-38.

Subotić A, Jevremović S, Trifunović M, Petrić M. Morpho-histological study of direct somatic embryogenesis in endangered species Fritillaria meleagris. Biol Plantarum. 2010;54:592-96.

Tchorbadjieva M. Protein markers for Somatic Embryogenesis. In: Mujib A, Šamaj J, editors. Plant Cell Monogr (2), Somatic Embryogenesis. Berlin Heidelberg: Springer Verlag; 2005. p. 215-33.

Tchorbadjieva M, Odjakova M. An acidic esterase as a biochemical marker for somatic embryogenesis in orchardgrass (Dactyilis glomerata L.) suspension cultures. Plant Cell Rep. 2001;20:28-33.

Burlina A, Galzigna L. A new and simple procedure for serum arylesterase. Clin Chim Acta. 1972;39:255-7.

Egertsdotter U, von Arnold S. Importance of arabinogalactan proteins for the development of somatic embryos of Norway spruce (Picea abies). Physiol Plantarum. 1995;93:334-45.

Van Engelen FA, De Vries SC. Secreted proteins in plant cell cultures. In: Roubelakis-Angelakis KA, Tran Than Van K, editors. Morphogenesis in plants. New York: Plenum Press; 1993. p. 181-200.

Balen B, Krsnik-Rasol M, Simeon-Rudolf M. Isoenzymes of peroxidase and esterase related to morphogenesis in Mammillaria gracilis Pfeiff. tissue culture. J Plant Physiol. 2003;160:1401-6.

Petrić M, Subotić A, Jevremović S, Trifunović-Momčilov M, Tadić V, Grujić M, Vujčić Z. Esterase and peroxidase isoforms in different stages of morphogenesis in Fritillaria meleagris L. in bulb-scale culture. C R Biol 2015;338:793-802.

Krsnik-Rasol M, Jelaska S, Šerman D. Isoperoxidases − early indicators of somatic embryoid differentiation in pumpkin tissue. Acta Bot Croat. 1982;41:33-9.

Joersbo M, Andersen JM, Okkels FT, Rajagopal R. Isoperoxidases as markers of somatic embryogenesis in carrot suspension cultures. Physiol Plantarum. 1989;76:10-6.

Balen B, Pavković D, Peharec P, Krnik-Rasol M. Biochemical markers of morphogenesis in long term horseradish (Armoracia lapathifolia Gilib.) tissue culture. Sci Hort. 2009;119:88-97.

Murashige T, Skoog F. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plantarum. 1962;15:473-97.

Bradford MM. A rapid sensitive method for the quantification of microgram quantities of proteins using the principle of protein-dye binding. Anal Biochem. 1976;72:248-54.

Vujčić Z, Lončar N, Dojnov B, Milovanović A, Vujčić M, Božić N. Characterisation of leucyl aminopeptidase from Solanum tuberosum tuber. Food Chem. 2010;121:418-23.

Siegel BZ, Galston AW. The isoperoxidases of Pisum sativum. Plant Physiol. 1976; 42:221-6.

Cavalcante Alves JM, Sihachakr D, Allot M, Tizroutine S, Mussio I, Servaes A, Ducreux G. Isozyme modifications and plant regeneration through somatic embryogenesis in sweet potato (Ipomoea batatas (L.) Lam). Plant Cell Rep. 1994;13:437-41.

Hilaire KT, Daouda K, Michel Z, Justin KY. Esterase isoenzymes are linked to embryogenic structure induction in cotton cell suspension cultures. Afr J Agr Res. 2007;2:394-8.

Mihaljević S, Radić S, Bauer N, Garić R, Mihaljević B, Horvat G, Ljeljak-Levanić D, Jelaska S. Ammonium-related metabolic changes affect somatic embryogenesis in pumpkin (Cucurbita pepo L.). J Plant Physiol. 2001;168:1943-51.

Winiarczyk K, Gębura J. Activity of selected hydrolytic enzymes in Allium sativum L. anthers. Plant Physiol Biochem. 2016;102:37-42.

Park JE, Park JY, Kim YS, Staswick PE, Jeon J, Yun J, Kim SY, Kim J, Lee JH, Park CM. GH3-mediated auxin homeostasis links growth regulation with stress adaptation response in Arabidopsis. J Biol Chem. 2007;282:10036-46.

Hrubcová M, Cvirkova M, Eder J. Peroxidase activities and content of phenolic acids in embryogenic and nonembryogenic alfalfa cell suspension cultures. Biol Plantarum. 1994;36:175-82.

Van Rossum MWPC, Alberda M, Van der Plas LHW. Role of oxidative damage in tulip bulb scale micropropagation. Plant Sci. 1997;130:207-16.


  • There are currently no refbacks.


Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.