Document Type : Research paper


1 Young Researchers Club, Isfahan (Khorasgan) Branch, Islamic Azad University, Isfahan, Iran

2 Department of plant protection, Faculty of Agriculture and Natural Resources, Isfahan (Khorasgan) Branch, Islamic Azad University, Isfahan, Iran


Cucumis melo L. is one of the most economically important horticultural crops worldwide. However,   low plant regeneration frequency of this plant during genetic transformation is  the  major hurdle for  applying biotechnological approaches. Hence, this study aimed to evaluate the effect of 6-benzyladenine (BA), cefotaxime (CTX), kanamycin (KAN), and indole-3-acetic acid (IAA) on the regeneration of cotyledonary petioles generated from 6-day-old in vitro grown seedlings. Results showed that  application of 1.5 mgl-1 BA plus 250 mgl-1 CTX and 1 mgl-1 BA with 1000 mgl-1 CTX formed the most efficient media for plant regeneration. The highest callus production was recorded on medium containing 1 mgl-1 BA with 250 mgl-1 CTX and 1.5 mgl-1 BA with 750 mgl-1 CTX. Medium containing 500 mgl-1 CTX plus 0.1 mgl-1 IAA  efficiently  induced both root \ and leaf formation. All regenerated plants were died by adding   100 mgl-1 kanamycin therefore this level considered as threshold level for kanamycin application.. Overall, the results indicated that presence of BA plays an essential role for melon regeneration and cefotaxime can be considered as an auxiliary agent. 


Abu-Romman S, Suwwan M, Al-Ramamneh E.A.D. 2013. The influence of plant growth regulators on callus induction from hypocotyls of cucumber (Cucumis sativus L.). Advances in Environmental Biology 7(1), 339-343.
Akasaka-Kennedy Y, Tomita K.O, Ezura H. 2004. Efficient plant regeneration and Agrobacterium-mediated transformation via somatic embryogenesis in melon (Cucumis melo L.). Plant Science 166(3), 763-769.
Choi J.Y, Shin J.S, Chung Y.S, Hyung N.I. 2012. An efficient selection and regeneration protocol for Agrobacterium-mediated transformation of oriental melon (Cucumis melo L. var. makuwa). Plant Cell, Tissue and Organ Culture 110(1), 133-140.
Chovelon V, Restier V, Giovinazzo N, Dogimont C, Aarrouf J. 2011. Histological study of organogenesis in Cucumis melo L. after genetic transformation: why is it difficult to obtain transgenic plants? Plant Cell Reports 30(11), 2001-2011.
Curuk S, Ananthakrishnan G, Singer S, Xia X, Elman C, Nestel D, Cetiner S, Gaba V. 2003. Regeneration in vitro from the hypocotyls of Cucumis species produces almost exclusively diploid shoots, and does not require light. HortScience 38(1), 105-109.
Curuk S, Elman C, Schlarman E, Sagee O, Shomer I, Cetiner S, Gray D.J, Gaba V. 2002. A novel pathway for rapid shoot regeneration from the proximal zone of the hypocotyl of melon (Cucumis melo L.). In Vitro Cellular & Developmental Biology - Plant 38(1), 260-267.
FAO. FAO Statistical Yearbook 2012: World Food and Agriculture. Food and Agricultural Organization of the United Nations, Rome. 2012. Available from: URL: (updated 4 August 2014)
Ficcadenti N, Rotino G.L. 1995. Genotype and medium affect shoot regeneration of melon. Plant Cell, Tissue and Organ Culture 40(3), 293-295
Guis M, Roustan J.P, Dogimont C, Pitrat M, Pech J.C. 1998. Melon biotechnology. Biotechnology & Genetic Engineering Reviews 15(1), 289-311.
Han S.N, Oh P.R, Kim H.S, Heo H.Y, Moon J.C, Lee S.K, Kim K.H, Seo Y.W, Lee B.M. 2007. Effects of Antibiotics on suppression of Agrobacterium tumefaciens and plant regeneration from wheat embryo. Journal of Crop Science and Biotechnology 10(2), 92-98.
Koné M, Koné T, Kouakou H.T, Konaté S, Ochatt J.S. 2013. Plant regeneration via direct shoot organogenesis from cotyledon explants of Bambara groundnut, Vigna subterranea L. Verdc. Biotechnology, Agronomy, Society and Environment 17(4), 584-592.
Liu E.E, Leung D.W, Xia Q.H, Zheng J.R, Peng X.X, He X.M. 2013. Efficient plant regeneration in vitro from cotyledon explants of chieh-qua (Benincasa hispida Cogn. var. chieh-qua). ScienceAsia. 39(2), 134-138.
Mathias R.J, Boyd L.A. 1986. Cefotaxime stimulates callus growth, embryogenesis and regeneration in hexaploid bread wheat (Triticum aestivum L. EM Thell). Plant Science 46(3), 217-223.
Murashige T, Skoog F. 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum 15(3), 473-497.
Naderi D, Mousavi A, Habashi A.A, Lotfi M. 2011. Optimization of somatic embryogenesis induction in Iranian melon (Cucumis melo cv. Khatooni). African Journal of Biotechnology 10(34), 6434-6438.
Nakano M, Mii M. 1993. Antibiotics stimulate somatic embryogenesis without plant growth regulators in several Dianthus cultivars.  Plant Physiology 141(6), 721- 725.
Nuňez-Palenius H.G, Cantliffe D.J, Huber D.J, Ciardi J, Klee H.J. 2006. Transformation of a muskmelon ‘Galia’ hybrid parental line (Cucumis melo L. var. reticulates Ser.) with an antisense ACC oxidase gene. Plant Cell Report 25(3), 198-205.
Paris H.S, Amar Z, Lev E. 2012. Medieval emergence of sweet melons, Cucumis melo (Cucurbitaceae). Annals of Botany 110(1), 23-33.
Rajagopalan P.A, Perl-Treves R. 2005. Improved cucumber transformation by a modified explant dissection and selection protocol. HortScience 40(2), 431-435.
Ren Y, Bang H, Curtis I.S, Gould J, Patil B.S, Crosby K.M. 2012. Agrobacterium-mediated transformation and shoot regeneration in elite breeding lines of western shipper cantaloupe and honeydew melons (Cucumis melo L.). Plant Cell, Tissue and Organ Culture 108(1), 147-158.
Rhimi A, Ben Fadhel N, Boussaid M. 2006. Plant regeneration via somatic embryogenesis from in vitro tissue culture in two Tunisian Cucumis melo cutivars Maazoum and Beji. Plant Cell, Tissue and Organ Culture 84(2), 239-243.
Silva J.D, Fukai S. 2001. The impact of carbenicillin, cefotaxime and vancomycin on chrysanthemum and tobacco TCL morphogenesis and Agrobacterium growth. Journal of Applied Horticulture 3(1), 3-12.
Souza F.V.D, Garcia-Sogo B, S.Souza A.D, San-Juán A.P, Moreno V. 2006. Morphogenetic response of cotyledon and leaf explants of melon (Cucumis melo L.) cv. Amarillo Oro. Brazilian Archives of Biology and Technology 49(1), 21-27.
Stipp L.C.L, Mendes B.M.J, Piedade S.M.S, Rodriguez A.P.M. 2001. In vitro morphogenesis of Cucumis melo var. inodorus. Plant Cell, Tissue and Organ Culture 65(1), 81-89.
Tabei Y, Kitade S, Nishizawa Y, Kikuchi K, Kayano T, Hibi T, Akutsu K. 1998. Transgenic cucumber plants harboring a rice chitinase gene exhibit enhanced resistance to gray mold (Botrytis cinerea). Plant Cell Report 17(3), 159-164.
Tang H, Ren Z, Krezal G. 2000. An evaluation of antibiotics for the elimination of Agrobacterium tumefaciens from walnut somatic embryos and for the effects on the proliferation of somatic embryos and regeneration of transgenic plants. Plant Cell Report 19(9), 881-887.
Venkateshwarlu M. 2012. Direct multiple shoots proliferation of muskmelon (Cucumis melo L.) From shoot tip explants. International Journal of Pharma and Bio Sciences 3(2), 645-652.
Wiebke BF, erreira, F, Pasquali G, Bodanese-Zanettini M. H, Droste A. 2006. Influence of antibiotics on embryogenic tissue and Agrobacterium tumefaciens suppression in soybean genetic transformation. Bragantia 65(4), 543-551.
Yepes L.M, Aldwinckle S. 1994. Factors that affect leaf regeneration efficiency in apple, and effect of antibiotics in morphogenesis. Plant Cell, Tissue and Organ Culture 37, 257-269.
Zhang H.J, Gao P, Wang X.Z, Luan F.S. 2014. An efficient regeneration protocol for Agrobacterium-mediated transformation of melon (Cucumis melo L.). Genetics and Molecular Research 13(1), 54-63.