Abstract
We report an efficient somatic embryogenesis and plant regeneration system using leaf cultures of Citrullus colocynthis (L.) and assessed the effect of plant growth regulators on the regeneration process. Initially leaf explants were cultured on Murashige and Skoog medium supplemented with different concentrations of auxins viz., 2,4-dichlorophenoxyacetic acid, 1-naphthaleneacetic acid, gibberellic acid alone and along with combination of 6-benzylaminopurine. The different forms of calli such as compact, white friable, creamy friable, brownish nodular, green globular and green calli were induced from the leaf explants on MS medium containing different concentrations of auxins and gibberellins. Subsequently initial callus was subcultured at 1.5 mg L−1 BAP + 1.0 mg L−1 2,4-D which resulted in 25 % somatic embryos from 85 % nodular embryogenic nodular callus that is highest percentage. Similarly the lowest percentage of somatic embryos was recorded at 2.5 mg L−1 BAP + 0.5 mg L−1 NAA from 55 % embryogenic globular callus i.e., 16 %. High frequency of embryo development takes place at intermittent light when compared with continuous light in the individual subcultures. The cotyledonary embryos were developed into complete platelets on MS medium. In vitro regenerated plantlets were washed to remove the traces of agar and then transferred to sterile vermiculite and sand (2:1) containing pot.
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References
Akasaka Kennedy Y, Tomita KO, Ezura H (2004) Efficient plant regeneration and Agrobacterium-mediated transformation via somatic embryogenesis in melon (Cucumis melo L.). PlantSci 166:763–769
Ali N, Skirvin RM, Splittstesser WE (1991) Regeneration of Cucumis sativus L. from cotyledons of small explants. HortScience 26:925–929
Cabrera-Ponce JL, Vegas-Garcia A, Herrera-Estrella L (1996) Regeneration of transgenic papaya plants via somatic embrogenesis induced by Agrobacterium rhizogenes. In vitro nCelll Dev Biol Plant 32:86–90
Cangahuala Inocente GC, Vesco LLD, Steinmacher D, Torres AC, Guerra MP (2007) Improvements in somatic embryogenesis protocol in Feijoa (Acca sellowiana (Berg) Burret): induction, conversion and synthetic seeds. Sci Hortic 111:228–234
Comlekcioglu N, Mendi YY, Eldogan S, Unek C (2009) Effects of different combinations and concentrations of growth regulators and photoperiod on somatic embryogenesis of Cucumis melo var. flexuosus. Afr J Biotechnol 8(22):6228–6232
Emmanuel S, Ignacimuthu S, Kathiravan K (2000) Micropropagation of Wedelia calendulacea less. A medicinal plant. Phytomorphology 50:195–200
Gonzalez JM, Friero E, Jouve N (2001) Infuence of genotype and culture medium on callus formation and plant regeneration from immature embryos of Triticum turgidum Dest. culture. Plant Breed 120:513–517
Herrington ME, Brown PJ, Carr AR (1996) Introgression as a source of bitterness in watermelon. HortScience 21:1237–1238
Junaid A, Mujib A, Bhat MA, Sharma MP, Samaj J (2007) Somatic embryogenesis and plant regeneration in Catharanthus roseus. Biol Plant 51:641–646
Kageyama K, Yabe K, Miyajima S (1991) Somatic embryogenesis in suspention culture of mature seed of Cucumis melo L. Jpn J Breed 41:273–278
Kim SG, Chang JR, Cha HC, Lee K (1988) Callus growth and plant regeneration in diverse cultivars of cucumber (Cucumis sativus L.). Plant Cell Tissue Organ Cult 12:67–74
Lakshmi SR, Benjamin JHF, Kumar TS, Murthy GVS, Rao MV (2013) Organogenesis from A in vitro-derived leaf and internode explants of A Hoya wightii ssp. palniensis—a vulnerable species of Western Ghats. Braz Arch Biol Technol 56:421–430
Maheswaran G, Williams EG (1986) Direct secondary somatic embryogenesis from immature embryos of Trifolium repens culture in vitro. Ann Bot 57:109–117
Martin KP (2004) Efficacy of different growth regulators at different stages of somatic embryogenesis Eryngium foetidum L. A rare medicinal plant. In vitro Cell Dev Biol Plant 40:459–463
Mohajer S, Taha RM, Khorasani A, Yaacob JS (2012) Induction of different types of callus and somatic embryogenesis in various explants of Sainfoin (Onobrychis sativa). Aust J Crop Sci 6(8):1305–1313
Murashige T, Skoog F (1962) A revised method for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:472–497
Noel M, Comeau D, Lenee P (1992) Embryogenese somatique de la courgette (Cucurbita pepo L.) a partir de cotyledons. In: Comptes-rendus du ler forum jeunes chercheurs del’IAPTC –‘Du gene a l’entreprise’. Amiens, France, pp 14–18
Novot N, Sarfatti M, Zamir D (1990) Linkage relationships of genes affecting bitterness andflesh color in watermelon. J Hered 81:162–165
Paramageetham Ch, Babu GP, Rao JVS (2004) Somatic embryogenesis in Centella asiatica L. An important medicinal and neutraceutical plant of India. Plant Cell Tissue Organ Cult 79:19–24
Ravishankar Rai V, Mc Comb J (2002) Direct somatic embryogenesis from mature embryos of sandalwood. Plant Cell Tissue Organ Cult 60:65–70
Rhimi A, Ben Fadhel N, Boussaid M (2006) Plant regeneration via somatic embryogenesis from in vitro tissue culture in two Tunisian Cucumis melocultivars Maazoun and Beji. Plant Cell Tissue Organ Cult 84:239–243
Shin H, Kim YW (2012) Somatic embryogenesis of Pinus rigida × P. taeda and the relationship between the initiation of embryogenic tissue and zygotic embryo development. Plant Biotechnol Rep 6:175–181
Thiruvengadam M, Jeyakumar JJ, Kamaraj M, Lee YJ, Chung IM (2013) Plant regeneration through somatic embryogenesis from suspension cultures of gherkin (Cucumis anguria L.). Aust J Crop Sci 7(7):969–977
Usman M, Hussain Z, Fatima B (2011) Somatic embryo genesis and shoot regeneration induced in cucumber leaves. Pak J Bot 43(2):1283–1293
Zlenko VA, Kotikov IV, Troshin LP (2002) Efficient GA3-assisted plant regeneration from cell suspensions of three grape genotypes via somatic embryogenesis. Plant Cell Tissue Organ Cult 70:295–299
Acknowledgments
The authors express their sincere thanks to the University Grants Commission, New Delhi, India, for financial support to carry out this work under major research project (UGC- MRP: F.No.41-530/2012) (SR). The authors are also, gratefully acknowledge to Prof A. Sadanandam, Dean, Faculty of Sciences, Kakatiya University, Warangal for his guidance in preparation of histological study of this article.
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Ramakrishna, D., Shasthree, T. High efficient somatic embryogenesis development from leaf cultures of Citrullus colocynthis (L.) Schrad for generating true type clones. Physiol Mol Biol Plants 22, 279–285 (2016). https://doi.org/10.1007/s12298-016-0357-z
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DOI: https://doi.org/10.1007/s12298-016-0357-z