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

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Authors

Stadwelson Dohling

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

e-mail:  staddohling@yahoo.co.in

Abstract

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.

Introduction

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

IAA

 

 

0.1

2.864±0.25d

3.996±0.23a

0.5

3.598±0.4c

3.066±0.10b

1.0

3.132±0.13cd

3.264±0.38ab

2.5

3.064±0.13cd

1.796±0.17c

5.0

2.464±0.20d

1.464±0.25c

BAP

 

 

0.1

2.864±0.17d

2.93±0.12cb

0.5

4.396±0.12b

2.73±0.13c

1.0

2.664±0.23d

1.864±0.17c

2.5

2.464±0.17d

1.198±0.08c

5.0

1.664±0.28e

-

BAP+IAA

 

 

0.1+0.1

2.33±0.28d

3.33±0.16ab

0.5+0.1

5.00±0.23a

3.88±0.30a

1.0+0.1

3.44±0.17c

3.22±0.22ab

0.1+0.5

2.11±0.20d

3.00±0.16b

0.5+0.5

2.44±0.24d

3.11±0.38ab

1.0+0.5

2.22±0.27d

3.00±0.28b

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)

Figures

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

References

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