Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

In vitro regeneration in Dierama erectum Hilliard

  • 244 Accesses

  • 17 Citations


The genus Dierama comprises plants with a potential to be developed as ornamentals. D. erectum seeds were decontaminated and germinated on 1/10th strength Murashige and Skoog (Physiol Plant 15:473–497, 1962) (MS) media without plant growth regulators or sucrose. In an experiment investigating the effects of 6-benzyladenine (BA), meta-Topolin (mT), kinetin (KIN) and zeatin (Z) with or without α-naphthaleneacetic acid (NAA), the highest shoot number per hypocotyl (4.20 ± 0.51) was obtained from MS medium supplemented with 1.0 μM Z after 8 weeks. This was followed by a combination of 2.0 μM KIN and 2.0 μM NAA with 3.67 ± 0.81 shoots per explant. BA treatments produced 3.20 ± 0.22 shoots per hypocotyl explant when 2.0 μM was combined with 1.0 μM NAA, while mT gave 3.09 ± 0.99 shoots per explant when 2.0 μM mT was combined with 2.0 μM NAA. Adventitious shoot regeneration was optimised when shoots were grown under a 16-h photoperiod at 100 μmol m−2 s−1 on MS medium supplemented with 1.0 μM BA. This resulted in an average of 12.73 ± 1.03 shoots per hypocotyl explant. Various concentrations of ancymidol, activated charcoal and sucrose did not promote in vitro corm formation of this species. Plants rooted successfully after 8 weeks on MS medium supplemented with 1.0 μM indole-3-butyric acid (IBA) and had an average root number of 2.73 ± 0.40. After 2 months of acclimatisation, plants had formed corms. The largest corms (of diameter 0.45 ± 0.03 cm) were produced in plants pre-treated with 0.5 μM IBA. The highest plant survival percentage of 73% was also associated with this treatment.

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2a–c
Fig. 3
Fig. 4a–e



Analysis of variance




Duncan’s multiple range test


Hydrochloric acid


Indole-3-acetic acid


Indole-3-butyric acid




Murashige and Skoog medium




α-naphthaleneacetic acid


Plant growth regulator


Potassium hydroxide


Sodium hypochlorite


Photosynthetic photon flux density




  1. Ascough GD, Erwin J, Van Staden J (2007) In vitro propagation of four Watsonia species. Plant Cell Tiss Organ Cult 88:135–145

  2. Ascough GD, Novák O, Pěnčik A, Rolčik J, Strnad M, Erwin JE, Van Staden J (2009) Hormonal and cell division analyses in Watsonia lepida seedlings. J Plant Physiol 166:1497–1507

  3. Bairu MW, Stirk WA, Doležal K, Van Staden J (2007) Optimizing the micropropagation protocol for the endangered Aloe polyphylla: can meta-topolin and its derivatives serve as replacement for benzyladenine and zeatin? Plant Cell Tiss Organ Cult 90:15–23

  4. Bairu MW, Kulkarni MG, Street RA, Mulaudzi RB, Van Staden J (2009) Studies on seed germination, seedling growth, and in vitro shoot induction of Aloe ferox Mill., a commercially important species. HortScience 44:751–756

  5. Bentley-Mowat JA, Reid SM (1968) Investigation of the radish leaf bioassay for kinetins, and demonstration of kinetin-like substances in algae. Ann Bot 32:23–32

  6. Bogaert I, Van Cauter S, Werbrouck SPO, Dolezal K (2006) New aromatic cytokinins can make the difference. Acta Hort 725:265–270

  7. Cabaleiro C, Economou AS (1991) Effect of light on rooting in vitro of Petunia microshoots. Acta Hort 300:189–195

  8. Daniel E (1997) The temperature dependence of photoinhibition in leaves of Phaseolus vulgaris (L.). Influence of CO2 and O2 concentrations. Plant Sci 124:1–8

  9. Dantu PK, Bhojwani SS (1995) In vitro corm formation and field evaluation of corm-derived plants of Gladiolus. Sci Hortic—Amsterdam 61:115–129

  10. De Klerk G-J, Ter Brugge J, Marinova S (1997) Effectiveness of indoleacetic acid, indolebutyric acid and naphthaleneacetic acid during adventitious root formation in vitro in Malus “Jork 9”. Plant Cell Tissue Organ Cult 49:39–44

  11. De Klerk G-J, Van Der Krieken W, De Jong JC (1999) The formation of adventitious roots: new concepts, new possibilities. In Vitro Cell Dev Biol Plant 35:189–199

  12. Economou AS, Read PE (1986) Influence of light duration and irradiance on micropropagation of a hardy deciduous azalea. J Am Soc Hort Sci 111:146–149

  13. Economou AS, Read PE (1987) Light treatments to improve efficiency of in vitro propagation systems. HortScience 22:751–754

  14. Fogaça CM, Fett-Neto AG (2005) Role of auxin and its modulators in the adventitious rooting of Eucalyptus species differing in recalcitrance. Plant Growth Regul 45:1–10

  15. Hilliard OM, Burtt BL (1991) Dierama: the hairbells of Africa. Acorn Books, Johannesburg

  16. Hussey G (1976) In vitro release of axillary shoots from apical dominance in monocotyledonous plantlets. Ann Bot 40:1323–1325

  17. Hutchings A, Scott AH, Lewis G, Cunningham AB (1996) Zulu medicinal plants: an inventory. University of Natal Press, Pietermaritzburg

  18. Kamínek M, Vaněk T, Motyka V (1987) Cytokinin activities of N6-benzyladenosine derivatives hydroxylated on the side-chain phenyl ring. J Plant Growth Regul 6:113–120

  19. Kamínek M, Motyka V, Vaňková R (1997) Regulation of cytokinin content in plant cells. Physiol Plant 101:689–700

  20. Madubanya LA, Makunga NP, Fennell CW (2006) Dierama luteoalbidum: liquid culture provides an efficient system for the ex situ conservation of an endangered and horticulturally valuable plant. S Afr J Bot 72:584–588

  21. Martin RC, Mok MC, Mok DWS (1999) A gene encoding the cytokinin enzyme zeatin O-xylosyltransferase of Phaseolus vulgaris. Plant Physiol 120:553–558

  22. McAlister BG, Jäger AK, Van Staden J (1998) Micropropagation of Babiana spp. S Afr J Bot 64:88–90

  23. Mok MC, Mok DWS, Armstrong DJ (1978) Differential cytokinin structure–activity relationships in Phaseolus. Plant Physiol 61:72–75

  24. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497

  25. Niimi Y, Onozawa T (1979) In vitro bulblet formation from leaf segments of lilies, especially Lilium rubellum Baker. Sci Hortic—Amsterdam 11:379–389

  26. Nissen SJ, Sutter EG (1990) Stability of IAA and IBA in nutrient medium to several tissue culture procedures. HortScience 25:800–802

  27. Page YM, Van Staden J (1985) In vitro propagation of Dierama latifolium. HortScience 20:1049–1050

  28. Palavan-Ünsal N, Çağ S, Çetin E, Büyüktunçer D (2002) Retardation of senescence by meta-topolin in wheat leaves. J Cell Mol Biol 1:101–108

  29. Pierik RLM, Steegmans HHM (1975) Effect of auxins, cytokinins, gibberellins, abscisic acid and ethephon on regeneration and growth of bulblets on excised bulb scale segments of hyacinth. Physiol Plant 34:14–17

  30. Plessner O, Ziv M, Negbi M (1990) In vitro corm production in the Saffron crocus (Crocus sativus L.). Plant Cell Tiss Organ Cult 20:89–94

  31. Pooley E (1998) A field guide to wild flowers of KwaZulu-Natal and the eastern region. Natal Flora Publications Trust, Durban

  32. Remotti PC (1995) Primary and secondary embryogenesis from cell suspension cultures of Gladiolus. Plant Sci 107:205–214

  33. Remotti PC, Löffler HJM (1995) Callus induction and plant regeneration from Gladiolus. Plant Cell Tiss Organ Cult 42:171–178

  34. Scott-Shaw CR (1999) Rare and threatened plants of KwaZulu-Natal and neighbouring regions. KwaZulu-Natal Nature Conservation Service, Pietermaritzburg

  35. Skoog F, Armstrong DJ (1970) Cytokinins. Annu Rev Plant Physiol 21:359–384

  36. Smulders MJM, Van De Ven ETWM, Croes AF, Wullems GJ (1990) Metabolism of 1-naphthaleneacetic acid in explants of tobacco: evidence for release of free hormone from conjugates. J Plant Growth Regul 9:27–34

  37. Takayama S, Misawa M (1982) Regulation of organ formation by cytokinin and auxin in Lilium bulbscales grown in vitro. Plant Cell Physiol 23:64–74

  38. Van Staden J, Crouch NR (1996) Benzyladenine and derivatives—their significance and interconversion in plants. Plant Growth Regul 19:153–175

  39. Watt JM, Breyer-Brandwijk MG (1962) The medicinal and poisonous plants of southern and eastern Africa: being an account of their medicinal and other uses, chemical composition, pharmacological effects and toxicology in man and animal. E. & S. Livingstone, London

  40. Werbrouck SPO, Strnad M, Van Onckelen HA, Debergh PC (1996) Meta-topolin, an alternative to benzyladenine in tissue culture? Physiol Plant 98:291–297

  41. Whitty CD, Hall RH (1974) A cytokinin oxidase in Zea mays. Can J Biochem 52:789–799

  42. Williams RR, Mungall ND, Taji AM (1992) Acclimatization: changes in photosynthetic competence. Acta Hort 314:131–136

Download references


We thank the National Research Foundation, Pretoria, and the National Manpower Development Secretariat (NMDS) of the Lesotho Government for their generous financial assistance. Our sincere gratitude also goes to Dr. Gary Stafford for helping with the seed collection.

Author information

Correspondence to J. Van Staden.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Koetle, M.J., Finnie, J.F. & Van Staden, J. In vitro regeneration in Dierama erectum Hilliard. Plant Cell Tiss Organ Cult 103, 23–31 (2010). https://doi.org/10.1007/s11240-010-9749-x

Download citation


  • Cytokinins
  • Dierama erectum
  • Micropropagation
  • Photoperiod
  • Threatened species