Effect of Culture Medium on In vitro Callus Induction of Guggul [Commiphora wightii (Arnott)] -An Endangered Medicinally Important Desert Plant

Effect of Culture Medium on In vitro Callus Induction of Guggul [Commiphora wightii (Arnott)] -An Endangered Medicinally Important Desert Plant

Swarnlata Kumawat^* , Mohan Lal Jakhar , Komal Shekhawat , Rekha Choudhary

Department of Plant Breeding and Genetics, SKN Agriculture University, Jobner, Jaipur, Rajasthan, India

Corresponding Author Email: kumawatswarnlata.sk@gmail.com

DOI : http://dx.doi.org/10.53709/CHE.2020.v01i01.021

Abstract

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Indian traditional system of medicine, guggul has been used for thousands of years to treat arthritis, inflammation, gout, rheumatism, obesity, and disorders of lipids metabolism [1]. Commiphora wightii (Arnott) is a medicinally valuable flowering plant now considered an endangered Burseraceae family species with chromosome number 2n = 26 [2]. Commiphora derives from the Greek words kommi (meaning ‘ gum ‘) and phero (meaning ‘ to bear ‘). It is known in the Indian languages by various names such as Hindi Guggul, Tamil Gukkulu and Maishakshi, Sanskrit Guggulu, and English Indian bdellium. The Commiphora genus is widespread in tropical regions of Africa, Madagascar, Asia, Australia and the Pacific Islands [3]. In India, it is found in arid, rocky tracts of Rajasthan and Gujarat, Maharashtra and Karnataka [4]. It is present in Rajasthan in Jaisalmer, Barmer, Jodhpur, Jalore, Sirohi, Ajmer, Sikar, Churu, Jhunjhunu, Pali, Udaipur, Alwar (Sariska Tiger Reserve), Jaipur (Ramgarh, Jhalana), Bhilwara and Rajsamand, among others. Commiphora wightii is a tiny shrub or tree. It is a slow – growing plant that takes 8 to 10 years to reach 3 to 3.5 meters high. The plant is dimorphic, one having male and bisexual flowers and the other having female staminodes flowers. The plant with only male flowers has also been reported [5]. The fruits are green berry-like drupe. The size of the fruit varies from 6 to 8 mm and 5 mm in diameter. Parts of fruit exposed to the sun develop a rosy tinge. Sowings reveal polyembryonic character [6].

            Guggul contains a fragrant gum resin called gugal, guggul or gugul, which is used in ayurvedic medicine in the incense and traditional remedy. Guggul gum is a mixture of 61 per cent resin, 29.3 per cent gum, 6.1 per cent water, 0.6 per cent volatile oil and 3.2 per cent foreign matter [7].

            Guggul was first presented to the scientific world in 1966 by G. V. Satyavati, an Indian Medical Researcher [8]. Guggul is considered to be endangered in India and is classified as ‘ Data Deficient ‘ in the IUCN Red Data List [9] due to a lack of information on its conservation status as well as excessive and unscientific taping methods to increase the yield of oleo-gum resin causes plant mortality leading to the species ‘ extinction. The wild population of this species has decreased by more than 80 per cent over the past 84 years (three-generation lengths) due to habitat loss and destruction, combined with uncontrolled harvesting and tapping of oleo-gum resin. Many species develop a fragrant oleo-gum-resin after damage to the bark [10]. Therefore this species is critically endangered [I1]. Over-exploitation, limited occurrence, small area of occupancy, severe fragmentation of populations, very low regeneration and invasion of alien species mean that Commiphora wightiiis facing a high extinction risk [12].

            Natural regeneration and cultivation are nearly negligible compared to their depletion. Slow growing nature associated with poor germination of the seeds is also the primary cause of Commiphora wightii. Efforts are required to preserve this species in nature and to propagate it extensively. The in vitro propagation method can be used for clonal propagation of selected germplasm, genetic improvement, production of active compounds in cell culture. Therefore, in vitro propagation method would be the promising option for multiplication and conservation of Commiphora wightii.

MATERIAL AND METHODS

            The present research was carried out at the Department of Plant Breeding and Genetics, S. K. N. College of Agriculture, Jobner, under the title “Effect of Culture Medium on In vitro Callus Induction of Guggul [ Commiphora wightii (Arnott) ] -An Endangered Medicinally Important Desert Plant.”

PLANT MATERIAL

A current investigation on guggul was conducted. Leaves were used as explants and obtained from herbal gardens kept under the Department of Plant Breeding and Genetics, S.K.N. Agricultural College, Jobner.

CULTURE MEDIUM

Various media such as Murashige and Skoog, Nitsch & Nitsch, Woody Plant Medium, Schenk and Hildebrant Medium, White’s Medium and Khudson Solution – C have been checked for callus induction at most sensitive plant growth regulator rates.

AUTOCLAVING

            The culture media in glass containers sealed with cotton plugs and covered with aluminium foils were autoclaved at 15 psi and 121 ⁰C for 15- 40 minutes. Exposure time depends on the volume of the liquid to be sterilized. After autoclaving, the media were stored in the dark for 48 hours at 25 + 2 ⁰C.

EXPLANT PREPARATION AND STERILIZATION

            Leaf Explants of guggul were washed thoroughly in running tap water for 20 minutes. Again, these were cleaned with liquid detergent (Rankleen) for 10 minutes with vigorous shaking. After washing with detergent, explants were again washed with running tap water to remove any trace of detergent for 5 minutes. Explants were surface sterilized with 0.1 per cent HgCl₂ in a laminar air flow cabinet. Leaves explants were sterilization for 1 minute. These were thoroughly washed four to five times with sterilized double distilled water and inoculated on the different culture media supplemented with callus induction at most responsive level of plant growth regulators.

INOCULATION OF EXPLANT

            After sterilization, the explants were inoculated on different culture media aseptically. For inoculation, explants were transferred to sizeable sterile glass petriplates with the help of sterile forceps under strict aseptic conditions.

 CULTURE CONDITIONS

All cultures were incubated at 25+ 2 ⁰C under fluorescent light in a 14: 10 hour’s photoperiod with light intensity of 3000-3500 lux.

RESULTS

           Effect of different culture media on induction of callus mainly six types of culture media (MS medium, Woody Plant medium, Nitsch and Nitsch medium, Schenk and Hildebrant medium, White^’s medium and khudson solution C medium) were used. Different culture media were supplemented with most responsive level of auxin (2.0 mg/l 2,4-D) of callus induction in leaf explants. When leaf explant was supplemented in woody medium callus induction was initiated within 20 days of incubation. Maximum light pinkish brown, semi-compact callus inductions were observed (1.02 g) at Woody Plant Medium with 100 per cent frequency followed by MS medium (0.92 g) light brown, compact callus with 100 per cent frequency and White^’s medium (Table 1. and Fig. 1). Minimum yellow, compact callus proliferation was observed (0.55 g) with 80 per cent frequency in Nitsch and Nitsch medium. In Schenk and Hildebrant medium and Khudson solution C medium) inhibit callus induction.

Discussion

The composition of growth medium is an essential factor affecting growth and morphogenesis of plant tissues. Plant tissue culture medium consists of macronutrients, micronutrients, vitamins, amino acids or other nitrogen supplements, carbon sources, organic supplements, solidifying agents and growth regulators. [13] the most commonly used medium in plant tissue culture. The B₅ [14], N₆ [15] and [16] have also been widely used for many plant species. Moreover, for culture of woody species, the DKW [17] and the WPM medium [18-19] are used. The growth medium is selected on the basis of purpose of tissue culture and for the plant species [20-22].

Perusal of Table 1 revealed that different callus induction was observed in various culture media at 2.0 mg/l 2, 4-D. Maximum callus induction was observed in Woody Plant medium with 100 per cent frequency. Similar observation was reported by [23] in pomegranate.

Table 1: Effect of different media supplemented with 2.0 mg/l 2,4-D on callus induction from leaf explant of guggul

S. No.	Media	Days taken for callus initiation	Colour of callus	Texture of callus	Morphogenetic response (%)	Fresh weight (mg)
1	Murashige and Skoog Medium	20.1	Light brown	Compact	100	928
2	Woody Plant Medium	19.3	Light pinkish brown	Semi-compact	100	1025
3	Nitsch and Nitsch Medium	22	Yellow	Compact	80	556
4	Schenk and Hildebrant Medium	–	–	–	–	–
5	White’s Medium	24	Yellow	Semi-compact	100	670
6	Khudson Solution-C	–	–	–	–	–

Conclusion

From the present study it is concluded that woody Plant medium supplemented with 2.0 mg/l 2,4-D was found best medium for callus induction in leaf explants than other media (MS medium, Woody Plant medium, Nitsch and Nitsch medium, Schenk and Hildebrant medium, White medium and Khudson solution C medium). Maximum light pinkish brown, semi-compact callus induction was observed in Woody Plant Medium with 100% frequency.

References

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