Plant Cell, Tissue and Organ Culture (PCTOC)

, Volume 139, Issue 1, pp 65–75 | Cite as

Sodium nitroprusside enhances callus induction and shoot regeneration in high value medicinal plant Canscora diffusa

  • Sivakumar SubiramaniEmail author
  • Sathish Sundararajan
  • Hari Priya Sivakumar
  • Venkatesh Rajendran
  • Sathishkumar RamalingamEmail author
Original Article


The present investigation was carried out to demonstrate the plant morphogenetic potential of sodium nitroprusside (SNP) in a commercially important medicinal plant Canscora diffusa. Leaf and nodal explants were used and highest callusing frequency (97.18%) was recorded for leaf explants supplemented with 1.0 mg/L NAA +2.0 mg/L 2ip + 15 µM SNP. SNP enhanced multiple shoot formation in both nodal and leaf explants derived callus considerably in the media supplemented with 1.0 mg/L NAA + 15 µM SNP and 0.5 mg/L BAP + 15 µM SNP (98.32 and 96.12%) respectively. The root induction also drastically increased with the supplementation of SNP (10 µM) in combination with 1.0 mg/L IBA. Our results proved that SNP with other growth hormones acts synergistically to enhance the in vitro responses including the callus, shoot, and root induction. This optimized tissue culture system will be very useful for the mass propagation of C. diffusa for both commercial and conservation aspects.

Key message

First report using sodium nitroprusside to enhance in vitro propogation of Canscora diffusa presented. Synergestic action of various growth regulators on SNP supplementation reported. Leaf and nodal segments used as explants produced better callusing and higher shoot formation with SNP treatments.


Sodium nitroprusside Canscora diffusa Plant growth regulators Mass multiplication 



Dr. S. Sivakumar thanks National Post-Doctoral Fellowship (Sanction Order No: PDF/2016/002258), Science and Engineering Research Board, Department of Science and Technology, Government of India. Mr. S. Sathish acknowledges ICMR, New Delhi (No.3/1/2/102/2018-Nut.) for fellowship support. We would also like to thank, University Grants Commission-Special Assistance Programme (UGC-SAP) and Department of Science and Technology-Fund for Improvement of S&T Infrastructure in Higher Educational Institutions (DST-FIST) for the financial support to carry out this research.

Author contributions

SS and SS designed, executed the experiments and prepared the manuscript. HPS and VR contributed substantially to the research in experimental and statistical analysis. RS mobilized the funds and critically evaluated the manuscript.

Compliance with ethical standards

Conflict of interest

All authors read, approved the manuscript and declare that there is no conflict of interest.

Supplementary material

11240_2019_1663_MOESM1_ESM.docx (348 kb)
Supplementary material 1 (DOCX 347 kb)


  1. Ahloowalia BS, Maluszynski M, Nichterlein K (2001) Induced mutation: a new paradigm in plant breeding. Euphytica 118:167–173CrossRefGoogle Scholar
  2. Barthel M (2008) U.S. Patent Application No. 11/666,177Google Scholar
  3. Beligni MV, Lamattina L (2002) Nitric oxide interferes with plant photo-oxidative stress by detoxifying reactive oxygen species. Plant Cell Environ 25:737–748CrossRefGoogle Scholar
  4. Canter PH, Thomas H, Ernst E (2012) Bringing medicinal plants into cultivation: opportunities and challenges for biotechnology. Trends Biotechnol 23:180–185CrossRefGoogle Scholar
  5. Carmona F, Pereira AMS (2013) Herbal medicines: old and new concepts, truths and misunderstandings. Rev Bras Farmacogn Braz J Pharmacogn 23:379–385CrossRefGoogle Scholar
  6. Chavan JJ, Nalawade AS, Gaikwad NB, Gurav RV, Dixit GB, Yadav SR (2014) An efficient in vitro regeneration of Ceropegia noorjahaniae: an endemic and critically endangered medicinal herb of the Western Ghats. Physiol Mol Biol Plants 20:405–410CrossRefGoogle Scholar
  7. Choudhuri RK, Ghoshal S (1971) Xanthones of Canscora Decussata Schult. Phytochem 10:2425–2432CrossRefGoogle Scholar
  8. Correa-Aragunde N, Graziano M, Lamattina L (2004) Nitric oxide plays a central role in determining lateral root development in tomato. Planta 218:900–905CrossRefGoogle Scholar
  9. Correa-Aragunde N, Graziano M, Chevalier C, Lamattina L (2006) Nitric oxide modulates the expression of cell cycle regulatory genes during lateral root formation in tomato. J Exp Bot 57:581–588CrossRefGoogle Scholar
  10. Dubey NK, Kumar R, Tripathi P (2004) Global promotion of herbal medicine: India’s opportunity. Curr Sci 86:37–41Google Scholar
  11. El-Beltagi HS, Ahmed OK, Hegazy AE (2015) Molecular role of nitric oxide in secondary products production in Ginkgo biloba cell suspension culture. Not Bot Horti Agrobot Cluj-Napoca 43(1):12–18CrossRefGoogle Scholar
  12. Gao Z, Lin Y, Wang X, Wang X, Wei M, Yang F, Shi Q (2012) Sodium nitroprusside (SNP) alleviates the oxidative stress induced by NaHCO3 and protects chloroplast from damage in cucumber. Afr J Biotechnol 11:6974–6982Google Scholar
  13. Ghosal S, Saini KS, Sinha BN (1983) Diffutin, a new adaptogenic glucosyloxyflavan from Canscora diffusa. J Chem Res Synopses 12Google Scholar
  14. Gouvêa CMCP, Souza JF, Magalhães CAN, Martins IS (1997) NO releasing substances that induce growth elongation in maize root segments. Plant Growth Regul 21:183–187CrossRefGoogle Scholar
  15. Han X, Yang H, Duan K, Zhang X, Zhao H, You S, Jiang Q (2009) Sodium nitroprusside promotes multiplication and regeneration of Malus hupehensis in vitro plantlets. Plant Cell Tiss Organ Cult 96:29–34CrossRefGoogle Scholar
  16. Huang AX, She XP (2003) Effect of SNP on rooting of hypocotyls cutting from mung bean seedling. Acta Bot Boreal-Occident Sin 23:2196–2199Google Scholar
  17. Jhanji S, Setia RC, Kaur N, Kaur P, Setia N (2012) Role of nitric oxide in cadmium-induced stress on growth, photosynthetic components and yield of Brassica napus L. J Environ Biol 33:1027–1032Google Scholar
  18. Kadota M, Niimi Y (2003) Effects of cytokinin types and their concentrations on shoot proliferation and hyperhydricity in in vitro pear cultivar shoots. Plant Cell Tiss Organ Cult 72:261–265CrossRefGoogle Scholar
  19. Kalra C, Babbar SB (2010) Nitric oxide promotes in vitro organogenesis in Linum usitatissimum L. Plant Cell Tiss Organ Cult 103:353–359CrossRefGoogle Scholar
  20. Kalra C, Babbar SB (2012) Stimulatory and period-specific effect of nitric oxide on in vitro caulogenesis in Albizzia lebbeck (L.) Benth. Acta Physiol Plant 34:387–392CrossRefGoogle Scholar
  21. Kaviani B (2014) Micropropagation of ten weeks (Matthiola incana) and lisianthus (Eustoma grandiflorum) (two ornamental plants) by using Kinetin (Kin), naphthalene acetic acid (NAA) and 2,4-dichlorophenoxyacetic acid (2,4-D). Acta Sci Pol Hortorum Cultus 13(1):141–154Google Scholar
  22. Khurana A, Khurana JP, Babbar SB (2011) Nitric oxide induces flowering in the duckweed Lemna aequinoctialis Welw. (Syn. L. paucicostata Hegelm.) under noninductive conditions. J Plant Growth Regul 30:378–385CrossRefGoogle Scholar
  23. Kogje K, Gahane R (2012) Studies on propagation, in vitro regeneration and commercial exploitation of Canscora decurrens Dalzell—a potential medicinal plant. Indian J Biotechnol 11:470–475Google Scholar
  24. Kohmura H, Araki H, Imoto M (1995) Micropropagation of ‘Yamatoimo’ Chinese yam (Dioscorea opposita) from immature leaves. Plant Cell Tiss Org Cult 40:271–276CrossRefGoogle Scholar
  25. Krasylenko YA, Yemets AI, Blume YB (2010) Functional role of nitric oxide in plants. Russ J Plant Physiol 57:451–461CrossRefGoogle Scholar
  26. Lamattina L, Garcia-Mata C, Graziano M, Pagnussat G (2003) Nitric oxide: the versatility of an extensive signal molecule. Annu Rev Plant Biol 54:109–136CrossRefGoogle Scholar
  27. Leshem YY (1996) Nitric oxide in biological systems. J Plant Growth Regul 18:155–169CrossRefGoogle Scholar
  28. Leterrier M, Valderrama R, Chaki M, Airaki M, Palma JM, Barroso JB, Corpas FJ (2012) Function of nitric oxide under environmental stress conditions. In: Khan NA, Nazar R, Iqbal N, Anjum NA (eds) Phytohormones and abiotic stress tolerance in plants. Springer, Berlin, pp 99–113CrossRefGoogle Scholar
  29. Madhavan V, Yoganarasimhan SN, Gurudev MR (2008) Pharmacognacy studies on Shankhpushp. (Convolvulusmicrophyllus, Sieb. ex Spreng. And Evolvulusalsinodes (L.) L. Indian J Trad know 7:529–541Google Scholar
  30. Malik SK, Chaudhary R, Kalia RK (2005) Rapid in vitro multiplication and conservation of Garcinia indica: a tropical medicinal three species. Sci Hort 106:539–553CrossRefGoogle Scholar
  31. Mitra R (1985) Bibliography on pharmacognosy of medicinal plants. National Botanical Research Institute, Lucknow, pp 549–551Google Scholar
  32. Mungole A, Day S, Kamble R, Kanfade H, Chaturvedi A, Zanwar P (2010) Active phytochemical and antibacterial potentiality of in vitro regenerated plantlets of Canscora decurrens (Dalzell). Indian J Sci Technol 3:679–683Google Scholar
  33. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:173–197CrossRefGoogle Scholar
  34. Nagasawa A, Finer JJ (1989) Plant regeneration from embryogenic suspension culture of Chinese yam (Dioscorea opposita Thunb.). Plant Sci 60:263–271CrossRefGoogle Scholar
  35. Ötvos K, Pasternak TP, Miskolczi P, Domoki M, Dorjgotov D, Szucs A, Bottka S, Dudits D, Feher A (2005) Nitric oxide is required for, and promotes auxin-mediated activation of, cell division and embryogenic cell formation but does not influence cell cycle progression in alfalfa cell cultures. Plant J 43:849–860CrossRefGoogle Scholar
  36. Pagnussat GC, Simontacchi M, Puntarulo S, Lamattina L (2002) Nitric oxide is required for root organogenesis. Plant Physiol 129:954–956CrossRefGoogle Scholar
  37. Pagnussat GC, Lanteri ML, Lamattina L (2003) Nitric oxide and cyclic GMP are messengers in the indole acetic acid-induced adventitious rooting process. Plant Physiol 132:1241–1248CrossRefGoogle Scholar
  38. Raskin I, Ribnicky DM, Komarnytsky S, Ilic N, Poulev A, Borisjuk N (2002) Plants and human health in the twenty-first century. Trends Biotechnol 20:522–531CrossRefGoogle Scholar
  39. Samsampour D, Sadeghi F, Asadi M, Ebrahimzadeh A (2018) Effect of nitric oxide (NO) on the induction of callus and antioxidant capacity of Hyoscyamus niger under in vitro salt stress. J Appl Bot Food Qual 91:24–32Google Scholar
  40. Sarropoulou V, Maloupa E (2017) Effect of the NO donor “sodium nitroprusside”(SNP), the ethylene inhibitor “cobalt chloride”(CoCl2) and the antioxidant vitamin E “α-tocopherol” on in vitro shoot proliferation of SideritisraeseriBoiss & Heldr. subsp. raeseri. Plant Cell Tiss Organ Cult 128(3):619–629CrossRefGoogle Scholar
  41. Sarropoulou V, Dimassi-Theriou K, Therios I (2015) Effect of sodium nitroprusside on micropropagation and biochemical parameters of CAB-6P and Gisela 6 cherry rootstocks. Turk J Biol 39(4):595–610CrossRefGoogle Scholar
  42. Schmidt BM, Ribnicky DM, Lipsky PE, Raskin I (2007) Revisiting the ancient concept of botanical therapeutics. Nat Chem Biol 3:360–366CrossRefGoogle Scholar
  43. Shahina PM, Nampy S (2014) A taxonomic revision of the genus Canscora in South India, and the erection of the new genus Canscorinella (Canscorinae, Gentianaceae) with two new combinations. Phytotaxa 164:201–225CrossRefGoogle Scholar
  44. Shekhawat MS, Kannan N, Manokari M (2015) In vitro propagation of traditional medicinal and dye yielding plant Morinda coreia Buch. Ham. S Af J Bot 100:43–50CrossRefGoogle Scholar
  45. Shirdel M, Motallebi-Azar A, Masiha S, Mortazavi N, Matloobi M, Sharafi Y (2011) Effects of inorganic nitrogen source and NH4+: NO3 ratio on proliferation of dog rose (Rosa canina). J Med Plants Res 5(18):4605–4609Google Scholar
  46. Sivanesan I, Song JY, Hwang SJ, Jeong BR (2011) Micropropagation of Cotoneaster wilsonii Nakai a rare endemic ornamental plant. Plant Cell Tiss Organ Cult 105(1):55–63CrossRefGoogle Scholar
  47. Struwe L (2014) Classification and evolution of the family Gentianaceae. Springer, BerlinCrossRefGoogle Scholar
  48. Sun BT, Jing Y, Chen KM, Song LL, Chen FJ, Zhang LX (2007) Protective effect of nitric oxide on iron deficiency-induced oxidative stress in maize (Zea mays). J Plant Physiol 164:536–543CrossRefGoogle Scholar
  49. Sung CH, Hong JK (2010) Sodium nitroprusside mediates seedling development and attenuation of oxidative stresses in Chinese cabbage. Plant Biotechnol Rep 4:243–251CrossRefGoogle Scholar
  50. Xu J, Yin H, Wang W, Mi Q, Liu X (2009) Effects of sodium nitroprusside on callus induction and shoot regeneration in micropropagated Dioscorea opposita. Plant Growth Regul 59(3):279CrossRefGoogle Scholar
  51. Yadav V (2016) Effect of gamma radiation on various growth parameters and biomass of Canscora decurrens Dalz. Int J Herb Med 4:109–115Google Scholar
  52. Yadav MK, Gaur AK, Garg GK (2003) Development of suitable protocol to overcome hyperhydricity in carnation during micropropagation. Plant Cell Tiss Organ Cult 72(2):153–156CrossRefGoogle Scholar
  53. Zamani M, Hakimi MH, Mosleh Arany A, Kiani B, Rashtian A (2014) The effects of salicylic acid (SA) and sodium nitroprusside (SNP) on physical and growth characteristics of Pinus eldarica. Bull Env Pharmacol Life Sci 3:31–35Google Scholar
  54. Zandonadi DB, Santos MP, Dobbss LB, Olivares FL, Canellas LP, Binzel ML, Okorokova-Façanha AL, Façanha AR (2010) Nitric oxide mediates humic acids-induced root development and plasma membrane H + -ATPase activation. Planta 231:1025–1036CrossRefGoogle Scholar

Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  • Sivakumar Subiramani
    • 1
    Email author
  • Sathish Sundararajan
    • 1
  • Hari Priya Sivakumar
    • 1
  • Venkatesh Rajendran
    • 1
  • Sathishkumar Ramalingam
    • 1
    Email author
  1. 1.Plant Genetic Engineering Laboratory, Department of BiotechnologyBharathiar UniversityCoimbatoreIndia

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