In vitro propagation of Hedychium gardnerianum Sheppard ex Ker Gawl., an important ornamental plant

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Stadwelson Dohling

Department of Botany, Lady Keane College, Shillong (Meghalaya) 793001, India



This study was conducted to develop an efficient protocol for mass propagation of Hedychium gardnerianum. Explants from rhizome buds were cultured on Murashige and Skoog (MS) medium supplemented with 6-Benzylaminopurine (BAP) and indole 3-acetic acid (IAA) alone (0.1 to 5 mg l-1) or a combination of BAP and IAA. MS medium supplemented with a combination of 0.5 mg l-1 BAP and 0.1 mg l-1 IAA produced the highest mean number of shoots (5) and highest number of roots (3.88) per explant as compared to other concentrations. Thus, combined effects of BAP and IAA improved significantly the shoot growth and proliferation. The proliferated shoots were green and healthy in appearance. Finally, healthy and complete plants with well developed roots were hardened, acclimatized and planted in the field successfully.

Keywords Hedychium gardnerianum, micropropagation, in vitro, acclimatization, ornamental plants.


Zingiberaceae is the largest family in the order Zingiberales with 53 genera and over 1200 species (Kress et al., 2002). Some plants of the family are rhizomatous herbs found throughout tropical and subtropical regions with its main distribution in Asia (Khatun et al., 2003). Hedychium is a genus of perennial plants belongs to Zingiberaceae native to tropical Asia and the Himalayas, commonly growing to between 1-2 m tall. Common names include garland flower, ginger lily, and kahili ginger. There are 50 species of Hedychium in tropical Asia (Kirtikar and Basu, 1984), 37 species in India and 8 species in Western Himalaya. Some species are grown as ornamental plants and one of the potential ornamental values is Hedychium gardnerianum.

H. gardnerianum commonly known as Kahili Ginger is an Eastern Himalaya plant which grows upto 2 m tall with long, bright green leaves clasping the tall stems. It has a fragrant, bright yellow flowers held in dense spikes (Fig. 1a). It has the typical sweet tropical scented gardenia like fragrance of the species but flowers much earlier and longer that is in the months of July-September and grows at an altitude of 4-8000 ft. It is one of the most popular ornamental Hedychium in North East India, but it is fast depleting and rare to find in the wild due to increasing deforestation and ruthless exploitation. Hence, it is necessary to develop methods for it mass propagation as there is no micropropagation procedure available for this species.

Materials and Methods

Newly sprouted rhizomatous shoots of H.gardnerianum were thoroughly washed in running tap water for 30 min along with few drops of surface disinfectant, Tween-20. These were then surface sterilized with 0.10% sodium hypochloride solution (4% available chlorine) for 5 min followed by 0.1% (w/v) mercuric chloride solution for 3 min. the explants were rinsed several time with sterile distilled water to removes all traces of surface disinfectant. Aseptic cultures were raised from embedded shoot bud of size 5-10 mm. Single explants was inoculated in 20×150 mm test tube containing sterilized MS (Murashige and Skoog, 1962) initiation medium supplemented with BAP (0.5 mg l-1) containing 3% (w/v) sucrose and 0.8% (w/v) agar. The pH of the medium was adjusted to 5.8, prior to autoclaving for 15 min at 1.06 kg cm-2 (121 oC). The cultures were incubated at a temperature of 23±2 oC under 12 h daily illumination with white fluorescent light of 50 µmoles-2 sec-1 intensity. The in vitro shoots obtained in the induction medium were used for further multiplication.

The rhizomatous shoots obtained from the induction medium were cut into 5-10 mm sizes and inoculated in MS medium supplemented with a range of growth regulators, viz., BAP and IAA singly and in combination (Table 1). Culture conditions and media used were similar to the induction medium. Observations were made on the shoot number and root number per explants after 4-5 weeks of culture. After rooting the complete plantlets were transferred to soil field pots for acclimatization. The data was subjected to statistical analysis using one way ANOVA and comparisons between the mean values of treatments were made by Fisher’s LSD test (Fisher, 1935).

Results and Discussion

The effect of BAP and IAA on in vitro induction of shoot and root of H. gardnerianum is summarized in Table 1. All the concentration used singly in MS medium showed varying response with the highest number of shoot was observed in BAP at the concentration of 0.5 mg l-1 with an average number of 4.39 shoots/explants (Fig. 1b), while the root number was highest in IAA at 0.1 mg l-1 with an average number of 3.99 roots/explants. It was observed with an increased of BAP alone in the medium the number of shoots and roots decreased which is in contrast to the finding in Zingiber zerumbet (Faridah et al., 2011). The role of BAP in shoots proliferation has been reported in other Zingiberaceae species (Ikeda and Tambe, 1989; Balachandran et al., 1990; Smith and Hamil, 1996; Rout et al., 2001; Panda et al., 2007; Mohanty et al., 2011; Abdelmageed et al., 2011).

The presence of BAP and IAA in the medium markedly improved the number of proliferating shoots. The results in combination for shoot elongation and rooting indicate that MS media with lower concentration of BAP and IAA (0.5 mg l-1 BAP+ 0.1 mg l-1 IAA) showed higher growth of shoots with an average of 5 shoots/explant and rooting with average of 3.88 roots/explants (Fig. 1c). Similar type of shoot growth from axillary bud was recorded from Hedychium spicatum (Koul et al., 2005) and Curcuma haritha, (Bejoy et al., 2006). Bejoy et al. (2006) also reported that the best shoot multiplication and root system were achieved on MS medium supplemented with 1.0 mg/l of BAP and 0.5 mg/l of IAA. This result confirmed this study, which indicated the importance of IAA hormone in the induction of roots. It was observed that cytokinin was required in optimal quantity for shoot proliferation in some species of Zingiberaceae, but inclusion of low concentration of auxins along with cytokinin triggered the rate of shoot proliferation (Rout and Das, 1997; Sharma and Singh, 1997). It was observed that IAA in combination with BAP promotes roots growth, while when IAA is used singly the higher the concentration there is a decrease in roots number. This result indicates that IAA promoted the growth of roots. Higher concentration of auxin, in the range that normally stimulates elongation of shoots, causes a significant inhibition of root growth (Hopkins and Hüner, 2004) Isolated long plantlets (Fig. 1d) were thoroughly washed with tap water and hardening of the well rooted plantlets was done in the potting mixtures of soil and sand (Fig. 1e, f) and kept under polyhouse condition for survival and growth. The present study therefore indicates that this protocol can be easily adopted for large scale cultivation of this species with higher multiplication rate.

Table 1. Effect of BAP and IAA on the morphogenetic response of H. gardnerianum

Growth regulators (mg l-1)

Mean no. of shoots/explants ±SE

Mean no. of roots/explants ±SE


























































Values are mean ± S.E. Means followed by same letter in the column are not significantly different as indicated by Fisher’s LSD (p = 0.05)


Fig 1:
a, Hedychium gardnerianum a closed view
b, multiple shoot formation in BAP (0.5 mg l-1)
c, multiple shoot formation and root formation in BAP (0.5 mg l-1) and IAA (0.1 mg l-1)
d, Isolated plants ready for transplantation
e,f, Hardened plantlets


  1. Abdelmageed, AHA., Faridah, QZ., Norhana, FMA., Julia, AA and Kadir, MA (2011) Micropropagation of Eltingera elatior by using axillary bud explants. J. Med. Plants Res. 5(13).
  2. Balachandran, SM., Bhat, SR and Chandel, KPS (1990) In vitro multiplication of tumeric (curcuma sp.) and ginger (Zingiber officinale Rosc.). Plant cell Rep. 8: 521-524.
  3. Bejoy, M., Dan, M and Anish, NP (2006) Factors affecting the in vitro multiplication of the endemic zingiber-Curcuma haritha Mangaly and Sabu.- Asian J. Plant Sci. 5: 847-853.
  4. Faridah, QZ., Abdelmageed, AHA., Julia, AA and Nor Hafizah, R (2011) Efficient in vitro regeneration of Zingiber zerumbet Smith (a valuable medicinal plant) plantlets from rhizome bud explants. Afri. J. Biotechnol. 10(46):9303-9308.
  5. Fisher, RA (1935) The Design of Experiment-Oliver and Boyd, Edinburgh-London.
  6. Hopkins, WG and Hüner, NP (2004). Introduction to Plant Physiology. 3rd Edn., John Wiley and Sons Inc., New York, pp. 576.
  7. Ikeda, IR and Tambe, MJ (1989) In vitro subculture application for ginger. Hort. Sci., 24: 142-143.
  8.  Kress, WJ., Prince, LM and Williams, KJ (2002) The Phylogeny and a New classification of ginger (Zingiberaceae): evidence from molecular data. Am. J. Bot. 89 (10): 1682-1696.
  9. Khatun, A., Nasrin, S and Hossain, MT (2003). Large scale multiplication of ginger (Zingiber officinale Rosc.) from shoot-tip culture. OnLine J. Biol. Sci. 3(1): 59-64.
  10. Kirtikar, KR and Basu, BD (1984). Indian medicinal plants. Bishan Singh, Mahendra Pal Singh, Dehradun. pp. 824-826.
  11. Koul, S., Raina, V and Sharma, SK (2005) Conservation and Propagation of High Altitude Medicinal and Aromatic Plant: Hedychium spicatum. J. Plant Biochem. & Biotech.14, 57-59.
  12. Mohanty, S., Panda, MK., Sahoo, S and Nayak, S (2011). Micropropagation of Zingiber rubens and assessment of genetic stability through RAPD and ISSR markers. Biol. Plant, 55(1): 16-20.
  13. Murashige, T and Skoog, F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant, 15: 473–497.
  14. Panda, MK., Mohanty, S., Subudhi, E., Acharya, L and Nayak, S (2007) Assessment of genetic stability of micropropagated plants of Curcuma longa L. by cytophotometry and RAPD analysis. Int. J. Integr. Biol. 1: 189-195.
  15. Rout, GR and Das, P (1997). In vitro organogenesis in ginger (Zingiber officinale Rosc.). J. Herbs, Spices Medicinal Plants, 4: 41-51.
  16. Rout, GR., Samantaray, S, and Das, P (2000). In vitro manipulation and propagation of medicinal plants. Biotechnol. Adv. 18: 91-120.
  17. Sharma, TR and Singh, BM (1997). High frequency in vitro multiplication of disease-free Zingiber officinale Rosc. Plant Cell Rep. 17: 68-72.
  18. Smith, MK and Hamil, SD (1996) Field evaluation of micro-propagated ginger in subtropical Queensland. Aust. J. Exp. Agric. 36: 347-354.

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