Effects of different growth harmones on shoot initiation of Vekhand (Acorus calamus L.) by using rhizomes

Introduction

The perennial herb known as the sweet flag, Acorus calamus L (also known as Bach), is a member of the Araceae family. On the northern temperate, subtropical, and hot regions of the Indian subcontinent, it can be seen in swampy soil, shallow water, and lake edges. Starch, resin (2.5%), volatile oil (1.5-3.5%), and tannins (1.5%) are all present. Asarone, one of calamus oil's primary ingredients. Acorus calamus is said to have antibacterial and antifungal properties in both its leaves and tuber. There has been a demand for the plant because of its many uses. Rarely does the herb yield seeds; instead, it is typically replicated vegetatively. The medical industry and the preservation of germplasm would both profit significantly from the in vitro techniques of clonal reproduction of Acorus calamus. Currently, using tissue culture technologies, micropropagation is a crucial assist in rapid clonal replication (Altaf et al., 2010) [1] . The Acorus calamus rhizome's crude extract is utilised in drinks and cosmetics. Seed production is scanty in this herb; hence, rhizomes are largely used as propagule for its traditional cultivation, which has a slow rate of micropropagation. As a result, in vitro procedures may offer a quicker method of mass micropropagation for the creation of highquality, disease-free quality seeds as well as a way to conserve genetic material (Quraishi et al., 2017) [7] . Conventional methods of micropropagating this plant is insufficient to supply the plant material needed for large-scale growth and to satisfy the expanding demand of the pharmaceutical companies, according to Hettiarchil et al (1997). As a result, new methods for micropropagating this plant species must be devised. One method for resolving these issues is plant multiplication through tissue culture. Plants that are genetically similar to their parent plants are produced by clonal propagation. Taking into account the value and viability of plant tissue culture technologies. Rhizome fragments are cultured in a manner that is limited to proliferation, differentiation, and regeneration, and this method has been employed for a wide range of therapeutic plant species (Nisa, 2018) [6] .

Material and Method

The research work was carried out in the Department of Plant Biotechnology, College of Agricultural Biotechnology, Loni, and Affiliated by MPKV, Rahuri. The Acorus calamus plant was identified and purchased from Dhanwantari Nursery of MPKV, Rahuri. The fresh Rhizomes were collected, washed thoroughly under running tap water for 15min to remove any adhering dirt, followed by distilled water, excised rhizome explants were washed with savolon soap solution for 10 min, followed with distilled water for 3 min. Then the washed explants were taken into aseptic room and rinsed with double distilled water for 3-4 times. Explants were surface disinfected with 0.1%(w/v) Aqueous mercuric chloride (HgCl 2 ) solution for 1min and rinsed with double distilled water for 3 to 4 times. And then soaked in 70% ethanol for 30 seconds. The surface sterilized rhizomes were cultured on MS medium sterilized by autoclaving at 121 0 C tempt, 15psi for 20 min. After inoculation all the cultures were grown under photo period of 16 hrs light (with white fluorescent tubes) at a temp of 25±3 0 C with different concentration and combinations of auxins for shoot formation. Data was recorded after 10 and 20 days of initial culture. The cultures having multiple shoots were excised and subcultured onto new medium. The MS medium 2.5 mg/l BAP + 1.0 mg/l IAA showed best growth in the Initiation stage therefore it was used again for the subculturing and multiplication stage. Similarly, for rooting, MS medium containing different concentrations of IBA and IAA from 1mg/l to 5 mg/l were first tested. Apart from single hormonal concentration, a combination of IBA and NAA was also tested for in vitro rooting of Acorus calamus.

Results and Discussion

Naturally grown, fresh rhizomes were cut into small pieces of about 1-3cm, were used as explants for shoot regeneration and multiplication. Regeneration of shoot from rhizome explant was observed at different hormone concentrations. The highest length of shoot was recorded at 2.5 mg/l BAP of around 8.125cm. and from the all other tested combinations of BAP+NAA and BAP+IAA, MS medium containing 2.5mg/l BAP and 1.0mg/l IAA showed highest average length of shoot of about 6.35cm

Conclusion

It can be stated that all tested media for initiation could be used for initiation of Vekhand. Different growth hormones were used in tissue culture of Acorus calamus with variable results for comparative studies. When rhizome segment was cultured on MS medium having concentration of 2.5 mg/l BAP, maximum shoot formation was obtained. The MS Media containing 2.5 mg/l BAP was further tested along with various combinations of NAA and IAA. The best growth was obtained in the MS media containing 2.5 mg/l BAP along with 1 mg/l IAA.

References

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