Document Type : Original Article


Botany Department, Agricultural and Biological Division, National Research Centre, Dokki, Giza, 33 El Bouhouth St., P.O. 12622, Egypt


Background: Tomato (Solanum lycopersicum L.) is an important plant rich in many vitamins and antioxidant enzymes.
Methods: Tomato leaves of two cultivars (‘Peto 86’ and ‘Strain B’) were used as explant sources for callus induction. The antioxidant activity of the calli ethanol (ET) and methanol (ME)extracts were determined. The enzymatic activities were evaluated in callus cultures. Callus induced from leaf explants of tomato cultivars on the Murashige and Skoog (MS) medium supplemented with various concentrations and combinations of cytokinins and auxins such as BAP (6-benzylaminopurine), NAA (1-naphthalene acetic acid), 2,4-D (2,4-dichlorophenoxyacetic acid), IAA (indole-3-acetic acid) and Kin (Kinetin) for rapid induction of callus and biomass growth.
Results: The medium (M2) containing 3 mg L-1 BAP with 1 mg L-1 IAA produced the highest percentage of callus induction (PCI) (100%) in two cultivars. The relative fresh weight growth (RFWG) was reported by the fresh callus weighed after four weeks of culture and again weighed after one month of sub-culture. In both cultivars cultured on M2 medium the RFWG was (1.60) in ‘Peto 86’ and (1.47) in ‘Strain B’. The results showed that PCI and RFWG differed with the cultivars tested. The scavenging DPPH free radical activity in callus (ET) extracts exhibited a significant increase in (P < 0.05) than the activity in callus (ME) extract. The peroxidase and polyphenol oxidase activities were found in calli of both tomato cultivars. The enzymatic activities were higher in callus of ‘Peto 86’ cultivar than in callus of ‘Strain B’ cultivar.
Conclusions: Calli had antioxidant and enzyme activities that can be beneficial for extracting important components or for plant regeneration.

Graphical Abstract

In vitro Callus Induction of Tomato and Evaluation of Antioxidant Activity of Aqueous Extracts and Enzymatic Activities in Callus Cultures


1. Raziuddin SS, Shah HJ, Chaudhary T, Mohammad AS. (2004). Hormonal effect on callus induction in tomato. Sarhad J. Agri., 20:223-225.
2. Wang RA, Zhang HB, Tanksley SD. (1994). Map based cloning in crop plants tomato as a model system, Genetic and Physical mapping of jointless. Mol. Gen. Genet., 242:681-688.  
3. Abd-Elrazig HE, Musa M, Elsheikh SE. (2018). Value chain analysis for tomato production and marketing in Khartoum state, Sudan. Curr. Inves. Agri. Curr. Res., 5(4):715-721. 
4. Mazyad HM, Khalil EM, Rezk AA, Abdel-Hakem MA, Aboul-Ata AE. (2007). Genetic studies on tomato yellow leaf curl begomovirus (TYLCV) resistance in Egypt: Six pooulation analysis. Int. J. Virol., 3:88-95.  
5. Bhatia P, Ashwath N, Senaratna T, Midmore D. (2004). Tissue culture studies of tomato(Lycopersicon esculentum). Plant Cell Tiss. Organ. Cult., 78:1-21
6. Kader A, Sinha SN, Ghosh P. (2015). Contribution of environmental factors on in vitro culture of an endangered and endemic mangroves Heritiera fomes Buch.-Ham. and Bruguiera gymnorhiza (L.) Lam. Trop. Plant Res., 2(3):192–203.
7. Tagi A, Kumar PP, Lakshmanan P. (2002). In Vitro Plant Breeding, Food Products Press, New York, pp 167.
8. Cardoza V. (2016). Tissue Culture: The manipulation of plant development. In Stewart Jr C.N. (Eds.), Plant Biotechnology and Genetics: Principles, techniques, and applications. New Jersey, NJ:John Wiley & Sons, pp. 113-132. 
9. Efferth T. (2018). Biotechnology applications of plant callus cultures. Engineering, 5(1):51-59.
10. Sohail AJ, Sabir HS, Shaukat A, Ghulam MA. (2015). The effect of plant growth regulators on callus induction and somatic embryogenesis of hybrid tomato. Pak. J. Bot., 47(5):1671-1677.
11. Shah SH, Ail S, Jan SA, Din J, Ali GM. (2015). Callus induction, in vitro shoot regeneration and hairy root formation by the assessment of various plant growth regulators in Tomato(Solanum lycopersicum Mill.). J. Animal Plant Sci., 25(2):528-538.
12. Lima JE, Benedito VA, Figueira A, Peres LEP. (2009). Callus, shoot and hairy root formation in vitro as affected by the sensitivity to auxin and ethylene in tomato mutants. Plant Cell Rep., 28:1169-1177.
13. Mostafiz SB, Waigran A. (2018). Efficient callus induction and regeneration in selected indica rice. Agronmy, 8:77.
14. Osman MG, Elhadi EA, Khalafalla MM. (2010). Callus formation and organogenesis of tomato(Lycopersicon esculentum Mill,C.V. Omdurman)induced by thidiazuron. African J. Biotechnol., 9(28):4407-4413.
15. Coenen C, Lomax TL. (1997). Auxin-cytokinin interactions in higher plants: old problems and new tools. Trends Plant Sci., 2(9):351-356.
16. Hanur VS, Krishnareddy B. (2016). In-vitro organogenesis in tomato (Solanum lycopersicum) using kinetin. Adv. Plant. Agric. Res., 4(6):397-401.
17. Umesh TG. (2014) In vitro callus induction and antioxidant potential of Decalepis hamiltonii (Wight and Arn). Int. J. Pharm. Sci., 6:452-456.
18. Pandey VP, Manika A, Swati S, Sameeksha T, Dwivedi UN. (2017). A comprehensive review on function and application of plant peroxidases. Biochem. Anal Biochem., 6:308.
19. Alhasnawi AN, Che-radziah CMZ, Kadhimi AA, Isahak A, Mohamad A, Yusoff WMW. (2016). Enhancement of antioxidant enzyme activities in rice callus by ascorbic acid under salinity stress. Biol. Plant., 60 (40):783-787.
20. Ullah AA, TungmunnithumD, Garros L, Drouet S, Hano C, Abbasi BH. (2019). Effect of ultraviolet-c radiation and melatonin stress on biosynthesis of antioxidant and antidiabetic metabolites produced in in vitro callus culture of Lepidium sativum L. Int. J. Mol. Sci., 20:1787.
21. Murashige T, Skoog F. (1962). A revised medium for rapid growth and bioassay with tobacco tissue cultures. Physiol. Plant, 15:473-497.
22. Chen JJ, Yue RQ Xu, HX Chen XJ. (2006). Study on plant regeneration of wheat mature embryos under endosperm supported culture. Agric. Sci. China, 5:572–578.
23. Johnson M, Wesely EG, Kavitha MS, Uma V. (2011). Antibacterial activity of leaves and inter-nodal callus extracts of Mentha arvensis L. Asia Pac. J. Sci. Technol., 4:196-200.
24. Liu CZ, Murrch SJ, El-Demerdash M, Saena PK. (2004). Artemisia judaica L.: micropropagation and antioxidant activity. J. Biotech., 110:63-71.
25. Kar M, Mishra D. (1975). Catalase, Peroxidase, and Polyphenoloxidase activities    during rice leaf senescence. Plant Physiol., 57:315-319.
26. Yousaf R, Khan MA, Ullah N, Khan I, Hayat O, Shehzad MA, Khan I, Taj F, Ud-Din N, Khan A, Naeem I, Ali H. (2019). Biosynthesis of anti-leishmanial natural products in callus cultures of Artemisia scoparia. Artif. Cell. Nanomed. Biotechn., 47(1):1122-1131.
27. Tanveer H, Ali S, Khan Z. (2012). Appraisal of secondary metabolites in in vitro cultures of Citrullu scolocynthis (L.). Schard Sci. Int. (Lahore), 24(1):75-80.
28. Jatoi SK, Sajid GM, Sappal H, Baloch MS, Qureshi A, Anwar R. (2001). Differential in vitro response of tomato hybrids against a multitude of hormonal regimes. J. Biol. Sci., 1:1141-1143.
29. Rzepka-Plevneš D, Kulpa D, Grabiec M, Kowalczys K, Kurek J. (2006). The Effect of Growth Regulators and Culture Conditions on the Callus Induction in Tomato. Acta Sci. Pol. Hortorum Cult., 5(2):23-34.
30. Sherkar HD, Chavan AM. (2014). Studies on callus induction and shoot regeneration in Tomato. Sci. Res. Rep., 4(1):89-93.
31. Reda E, Moghaieb A, Sneak H, Fujita K. (2004). Shoot regeneration GUS transformed tomato (Lycopersicon esculentum) hairy root. Cell Mol. boil., 9:439-449.
32. Abd El-Hameid, AR, Abo El-kheir, Z, Abdel-Hady, M, Helmy, W. (2018). In vitro and molecular characterization using ISSR markers of Glycyrrhiza glabra L. BioTechnologia, 99(4):409-416.
33. Rossignol M, Santoni V, Zponanski S, Wand-Vansuyt G. (1990). Differential sensitivity to auxin at the plasma membrane level. In: NijKamp, HJJ, Van Der Plas, LHW, Van Aartrijk, J, (eds.) Progress in Plant Cellular and Molecular Biology. Kluwer Academic Publisher, Dordrecht, The Netherlands,  pp 498-503.
34. Swamy MK, Mohanty SK, Anuradha M. (2014). The effect of plant growth regulators and natural supplements on in vitro propagation Pogostemon cablin Benth. J. Crop Sci. Biotech., 17(2):71-78.
35. Ghasemi S, Koohi DE, Emmamzadehhashemi MSB, Khamas SS, Moazen M, Hashemi AK, Amin G, Golfakhrabadi F, Yousefi Z, Yosefbeyk F. (2018). Investigation of phenolic compounds and antioxidant activity of leaves extracts from seventeen cultivars of Iranian olive(Olea europaea L.). J. Food Sci. Technol., 55(11):4600-4607.
36. Liu S, Liu L, Tang Y, Shuo X. (2017). Comparative transcriptomic analysis of key genes involved in flavonoid biosynthetic pathway and identification of a flavonol synthase from Artemisia annua L. Plant Biol. J., 19:618-629.
37. Abbasi BH, Khan MA, Mahmood T, Ahmad M, Chaudhary MF, Khan MA. (2010). Shoot regeneration and free-radical scavenging activity in Silybum marianum L. Plant Cell. Tiss. Organ. Cult., 101:371-376.
38. Kim HS, Chin KB. (2017). Evaluation of antioxidative activity of various levels of ethanol extracted tomato powder and application to pork patties. Korean J. Food Sci. Anim Resour., 37(2):242-253. 
39. Boonsiri K, Saichol K, Van-Doorn WG. (2007). Seed browning of hot peppers during low temperature storage. Postharvest Biol. Technol., 45:358-365.
40. Passardi F, Penel C, Dunand C. (2004). Performing the paradoxical: how plant peroxidases modify the cell wall. Trend Plant Sci., 9:534-40.  
41. Oksman-Caldentey KM, Barz WH. (Eds.) (2002). Plant Biotechnology and Transgenic Plants. (1 Ed.) CRC Press, pp. 720.
42. Elimasni S D, Rosmayati S L. (2016). In vitro resistant-induction of Tamarillo (Solanum betaceum Cav.) applied by UV-B radiation against anthracnose disease by determination of peroxidase and polyphenol oxidase activity. Inter. J. Sci. Tech. Res., 5(6):26-30.
43. Moreno OAV, Vazquez-Duhalt R, Ochoa JL. (1989). Peroxidase activity in calluses and cell suspension cultures of radish Raphanus sativus var.Cherry Bell. Plant Cell Tiss. Org. Cult., 18:321-327.
44. Khalid A, Aftab F. (2020). Effect of exogenous application of IAA and GA3 on growth, protein content, and antioxidant enzymes of solanum tuberosum L. grown in vitro under salt stress. In vitro Cell. Dev. Biol. Plant, 56:377–389.