A comparative study on the effects of ultrasound and some growth factors on somatic embryogenesis and artificial seed production in cucumber (Cucumis sativus L.)


  • Morvarid KOOCHANI Islamic Azad University, North Tehran Branch, Faculty of Biological Sciences, Department of Biology, 10th Bostan St., Southern Makran St., Heravi Sq., Tehran (IR)
  • Ahmad MAJD Islamic Azad University, North Tehran Branch, Faculty of Biological Sciences, Department of Biology, 10th Bostan St., Southern Makran St., Heravi Sq., Tehran (IR) https://orcid.org/0000-0003-3707-7581
  • Sedigheh ARBABIAN Islamic Azad University, North Tehran Branch, Faculty of Biological Sciences, Department of Biology, 10th Bostan St., Southern Makran St., Heravi Sq., Tehran (IR)
  • Faezeh GHANATI Tarbiat Modares University (TMU), Faculty of Biological Sciences, Department of Plant Biology, No.7, Jalal AleAhmad St., Tehran (IR)
  • Sayeh JAFARI MARANDI Islamic Azad University, North Tehran Branch, Faculty of Biological Sciences, Department of Biology, 10th Bostan St., Southern Makran St., Heravi Sq., Tehran (IR)




acclimatization; callogenesis; histology; sodium alginate; synthetic seeds


Cucumber (Cucumis sativus L.), a popular vegetable crop of the family Cucurbitaceae, is cultivated in different parts of the world and is of great economic importance. Ultrasound is known as a physical stimulus that can affect living systems. At the right frequency and exposure period, these waves have desired effects. In the present study, 2-week-old plants, were put in an ultrasonic bath with a nominal frequency of 40 kHz, a central frequency of 34/722 kHz and a bandwidth of 320 Hz for 0, 5, 10 and 15 minutes. Hypocotyl pieces were used as explants and cultured in Murashige and Skoog medium supplemented with 0/5 mg/l each of 2,4-dichlorophenoxyacetic acid, 1-Naphthalene acetic acid, kinetin and 6-benzylaminopurine. The use of ultrasonication at a nominal frequency of 40 kHz for 10 minutes showed better results as compared with the samples treated for 0, 5 and 15 minutes, in terms of the shape, maturity of the embryos, and also the germination of artificial seeds and maturing to flowering stage. Since the cucumber of Esfahan is very famous for its aroma and taste and it is somewhat at risk of extinction, we studied the production of artificial seeds of this valuable plant using somatic embryos and apical buds. This research is the first report on investigation of the positive effect of ultrasound on somatic embryogenesis and artificial seed production. our results clearly showed that this method is a practical method to accelerate seed germination and flowering stage.


Abu-Romman S, Suwwan M, Ezz A (2013). The influence of plant growth regulators on callus induction from hypocotyls of cucumber (Cucumis sativus L.). Advances in Environmental Biology 339-344.

Alam M, Amin R, Uddin M, Biswas S, Islam M (2015). Regeneration of shoot from nodal explants of Cucumis sativus considering different hormonal concentration. International Research Journal of Biological Sciences 4(7):48-52.

Bhatia S, Dahiya R (2015). Concepts and techniques of plant tissue culture science. Modern Applications of Plant Biotechnology in Pharmaceutical Sciences 121-126.

Bose S, Karmakar J, Fulzele DP, Basu U, Bandyopadhyay TK (2017). In vitro shoots from root explant, their encapsulation, storage, plant recovery and genetic fidelity assessment of Limonium hybrid ‘Misty Blue’: a florist plant. Plant Cell, Tissue Organ Culture 129(2):313-324. https://doi.org/10.1007/s11240-017-1179-6.

Cade RM, Wehner TC, Blazich FA (1990). Somatic embryos derived from cotyledons of cucumber. Journal of the American Society for Horticultural Science 115(4):691-696. https://doi.org/10.21273/JASHS.115.4.691.

Chen B, Huang J, Wang J, Huang L (2008). Ultrasound effects on the antioxidative defense systems of Porphyridium cruentum. Colloids Surf B Biointerfaces 61(1):88-92. https://doi.org/10.1016/j.colsurfb.2007.07.009.

Cheruvathur MK, Kumar GK, Thomas TD (2013). Somatic embryogenesis and synthetic seed production in Rhinacanthus nasutus (L.) Kurz. Plant Cell, Tissue Organ Culture 113(1):63-71. https://doi.org/10.1007/s11240-012-0251-5.

Da Silva JAT, Dobránszki J (2014). Sonication and ultrasound: impact on plant growth and development. Plant Cell, Tissue Organ Culture 117(2):131-143. https://doi.org/10.1007/s11240-014-0429-0.

Das D, Rahman A, Kumari D, Kumari N (2016). Synthetic seed preparation, germination and plantlet regeneration of litchi (Litchi chinensis Sonn.). American Journal of Plant Sciences 7(10):1395. http://dx.doi.org/10.4236/ajps.2016.710133.

Dobránszki J, da Silva JAT (2010). Micropropagation of apple-a review. Biotechnology Advances 28(4):462-488. https://doi.org/10.1016/j.biotechadv.2010.02.008

Duncan DB (1955). Multiple range and multiple F tests. Biometrics 11(1):1-42. https://doi.org/10.2307/3001478

Elmeer KMS, Hennerty MJ (2008). Observations on the combined effects of light, NAA and 2, 4-D on somatic embryogenesis of cucumber (Cucumis sativus) hybrids. Plant Cell, Tissue Organ Culture 95(3):381-384. https://doi.org/10.1007/s11240-008-9439-0.

Gaba V, Kathiravan K, Amutha S, Singer S, Xiaodi X, Ananthakrishnan G (2008). The uses of ultrasound in plant tissue culture. In: Plan Tissue Culture Engineering. Springer, Dordrecht pp 417-426.


Gantait S, Kundu S, Ali N, Sahu NC (2015). Synthetic seed production of medicinal plants: a review on influence of explants, encapsulation agent and matrix. Acta Physiologiae Plantarum, 37(5), 98. https://doi.org/10.1007/s11738-015-1847-2.

George EF (1993). Plant propagation by tissue culture. Part 1: The technology (No. Ed. 2). Exegetics limited.

Ghanati F, Safari M, Hajnorouzi A (2015). Partial clarification of signaling pathway of taxanes increase biosynthesis by low intensity ultrasound treatment in hazel (Corylus avellana) cells. South African Journal of Botany 96:65-70. https://doi.org/10.1016/j.sajb.2014.10.012.

Gopi C, Ponmurugan P (2006). Somatic embryogenesis and plant regeneration from leaf callus of Ocimum basilicum L. Journal of Biotechnol 126(2):260-264. https://doi.org/10.1016/j.jbiotec.2006.04.033.

Haque SM, Ghosh B (2017). Regeneration of Cytologically Stable Plants Through Dedifferentiation, Redifferentiation, and Artificial Seeds in Spathoglottis plicata Blume.(Orchidaceae). Horticultural Plant Journal 3(5):199-208. https://doi.org/10.1016/j.hpj.2017.10.002

Hassanein AM (2003). Somatic embryogenesis of cucumber (Cucumis sativus L) using seed cuttings obtained from pre-mature fruit. Plant Biotechnol (Tsukuba) 20(4):275-281. https://doi.org/10.5511/plantbiotechnology.20.275.

Isah T (2015). Adjustments to in vitro culture conditions and associated anomalies in plants. Acta Biologica Cracoviensia Series Botanica 57(2):9-28. https://doi.org/10.1515/abcsb-2015-0026.

Ju H-J, Jeyakumar J, Kamaraj M, Praveen N, Chung I-M, Kim S-H, Thiruvengadam M (2014). High frequency somatic embryogenesis and plant regeneration from hypocotyl and leaf explants of gherkin (Cucumis anguria L.). Scientia Horticulturae 169:161-168. https://doi.org/10.1016/j.scienta.2014.02.023.

Juturu VN, Mekala GK, Kirti P (2015). Current status of tissue culture and genetic transformation research in cotton (Gossypium spp.). Plant Cell, Tissue Organ Culture 120(3):813-839. https://doi.org/10.1007/s11240-014-0640-z.

Liu Y, Yoshikoshi A, Wang B, Sakanishi A (2003). Influence of ultrasonic stimulation on the growth and proliferation of Oryza sativa Nipponbare callus cells. Colloids Surf B Biointerfaces 27(4):287-293. https://doi.org/10.1016/S0927-7765(02)00052-8.

Lou H, Kako S (1995). Role of high sugar concentrations in inducing somatic embryogenesis from cucumber cotyledons. Scientia Horticulturae 64(1-2):11-20. https://doi.org/10.1016/0304-4238(95)00833-8.

Mamdouh D, Marghany HA, Ewais EA (2014). Assessment of somaclonal variation of calli and regenerated plants of three cucumber (Cucumis sativus L.) cultivars using molecular markers. Report and Opinion 6(12).

Mashayekhi K, Sharifani M, Shahsavand M, Kalati H (2012). Induction of somatic embryogenesis in absence of exogenous auxin in cucumber (Cucumis sativus L.). International Journal of Plant Production 2(2):163-166.

Micheli M, Standardi A (2016). From somatic embryo to synthetic seed in Citrus spp. through the encapsulation technology. In: In Vitro Embryogenesis in Higher Plants, Humana Press, New York pp 515-522. https://doi.org/10.1007/978-1-4939-3061-6_30

Murashige T, Skoog F (1962). A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiologia Plantarum 15(3):473-497. https://doi.org/10.1111/j.1399-3054.1962.tb08052.x.

Nowacka M, Wedzik M (2016). Effect of ultrasound treatment on microstructure, colour and carotenoid content in fresh and dried carrot tissue. Applied Acoustics 103:163-171. https://doi.org/10.1016/j.apacoust.2015.06.011.

Passalacqua N, Guarrera P, De Fine G (2007). Contribution to the knowledge of the folk plant medicine in Calabria region (Southern Italy). Fitoterapia 78(1):52-68. https://doi.org/10.1016/j.fitote.2006.07.005.

Rajewska K, Mierzwa D (2017). Influence of ultrasound on the microstructure of plant tissue. Innovative Food Science and Emerging Technologies 43:117-129. https://doi.org/10.1016/j.ifset.2017.07.034.

Raju CS, Kathiravan K, Aslam A, Shajahan A (2013). An efficient regeneration system via somatic embryogenesis in mango ginger (Curcuma amada Roxb.). Plant Cell, Tissue Organ Culture 112(3):387-393. https://doi.org/10.1007/s11240-012-0244-4.

Redenbaugh K, Paasch BD, Nichol JW, Kossler ME, Viss PR, Walker KA (1986). Somatic seeds: encapsulation of asexual plant embryos. Nature Biotechnology 4(9):797. https://doi.org/10.1038/nbt0986-797.

Rezaei A, Ghanati F, Behmanesh M, Mokhtari-Dizaji M (2011). Ultrasound-potentiated salicylic acid–induced physiological effects and production of taxol in hazelnut (Corylus avellana L.) cell culture. Ultrasound in Medicine and Biology 37(11):1938-1947. https://doi.org/10.1016/j.ultrasmedbio.2011.06.013.

Safari M, Ghanati F, Behmanesh M, Hajnorouzi A, Nahidian B, Ghahremani M (2013). Enhancement of antioxidant enzymes activity and expression of CAT and PAL genes in hazel (Corylus avellana L.) cells in response to low-intensity ultrasound. Acta Physiologiae Planterum 35(9):2847-2855. https://doi.org/10.1007/s11738-013-1318-6.

Sharififar A, Nazari M, Asghari HR (2015). Effect of ultrasonic waves on seed germination of Atriplex lentiformis, Cuminum cyminum, and Zygophyllum eurypterum. Journal of Applied Research on Medicinal and Aromatic Plants, 2(3 ), 102-104. https://doi.org/10.1016/j.jarmap.2015.05.003.

Smith RH (2012). Plant tissue culture: techniques and experiments. Academic Press.

Sunandar A, Supena EDJ (2017). Induction of somatic embryogenesis in sengon (Falcataria moluccana) with thidiazuron and light treatments. HAYATI Journal of Biosciences 24(2):105-108. https://doi.org/10.1016/j.hjb.2017.08.002.

Tabassum B, Nasir IA, Farooq AM, Rehman Z, Latif Z, Husnain T (2010). Viability assessment of in vitro produced synthetic seeds of cucumber. African Journal of Biotechnology 9(42):7026-7032.

Thorpe TA (2007). History of plant tissue culture. Molecular Biotechnology 37(2):169-180. https://doi.org/10.1007/s12033-007-0031-3.

Tripathi L, Tripathi JN (2003). Role of biotechnology in medicinal plants. Tropical Journal of Pharmaceutical Research 2(2):243-253. https://doi.org/10.4314/tjpr.v2i2.14607

Ugandhar T, Venkateshwarrlu M, Begum G, Srilatha T, Jaganmohanreddy K (2011). In Vitro plant regeneration of cucumber (Cucumis sativum (L.) from cotyledon and hypocotyl explants. Science Research Reporter 1(3):164-169. https://doi.org/10.13140/RG.2.1.4203.3366

Usman M, Hussain Z, Fatima B (2011). Somatic embryogenesis and shoot regeneration induced in cucumber leaves. Pakistan Journal of Botany 43(2):1283-1293.

Vasil IK (2008). A history of plant biotechnology: from the cell theory of Schleiden and Schwann to biotech crops. Plant Cell Reports 27(9):1423. https://doi.org/10.1007/s00299-008-0571-4.

Von Arnold S, Sabala I, Bozhkov P, Dyachok J, Filonova L (2002). Developmental pathways of somatic embryogenesis. Plant Cell, Tissue Organ Culture 69(3):233-249. https://doi.org/10.1023/A:1015673200621.

Yang H, Gao J, Yang A, Chen H (2015). The ultrasound-treated soybean seeds improve edibility and nutritional quality of soybean sprouts. Food Research International 77 704-710. https://doi.org/10.1016/j.foodres.2015.01.011.




How to Cite

KOOCHANI, M., MAJD, A., ARBABIAN, S., GHANATI, F., & JAFARI MARANDI, S. (2020). A comparative study on the effects of ultrasound and some growth factors on somatic embryogenesis and artificial seed production in cucumber (Cucumis sativus L.). Notulae Botanicae Horti Agrobotanici Cluj-Napoca, 48(4), 1915–1928. https://doi.org/10.15835/nbha48411669



Research Articles
DOI: 10.15835/nbha48411669

Most read articles by the same author(s)