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(Commiphora Wightii (Arnott)) Under Invitro Conditions Int.J.Curr.Microbiol.App.Sci (2021) 10(03): 1896-1902 International Journal of Current Microbiology and Applied Sciences ISSN: 2319-7706 Volume 10 Number 03 (2021) Journal homepage: http://www.ijcmas.com Original Research Article https://doi.org/10.20546/ijcmas.2021.1003.239 Influence of Antioxidants on the Explants of Guggul (Commiphora wightii (Arnott)) under invitro Conditions K. Shekhawat1*, S. Kumawat1, R. Kumari2 and M. L. Jakhar1 1Department of Plant Breeding and Genetics, S. K. N. Agriculture University, Jobner, Rajasthan, India 2Department of Plant Breeding and Genetics, Bihar Agriculture University, Sabour, Bhaglpur-813210, Bihar, India *Corresponding author ABSTRACT Shoot apex and nodal segment explants of guggul were placed on K eyw or ds Murashige and Skoog Medium (MS medium, 1962) supplemented with different concentration of activated charcoal (20, 40, 60, 80 and 100 mg/l) Guggul, Callus induction, Tissue and Polyvinylpyrrolidone (2, 4, 6, 8 and 10 mg/l) for shoot bud induction. culture, in vitro Activated charcoal (20 - 100 mg/l) was added in the basal medium with responsive level of plant growth regulators (3.0 mg/l BAP) in nodal Article Info segment, maximum number of shoot bud (1.55) was observe at 60 mg/l Accepted: activated charcoal with no browning intensity in the culture medium. 18 February 2021 Available Online: Maximum shoot bud proliferation in nodal segment explants followed by 6 10 March 2021 mg/l level of polyvinylpyrrolidone. Shoot apex explant was gave minimum response in activated charcoal and polyvinylpyrrolidone. Introduction the family Burseraceae and having the chromosome number 2n = 26 (Sobti and India has a treasure of well recorded and Singh, 1961) (4). It is an important medicinal traditionally well practiced knowledge on plant of herbal heritage of India (5). It is medicinal plants (1). More than 6000 plants known by various names like guggul in Hindi, are used in our traditional, folk and herbal gukkulu and maishakshi in Tamil, guggulu in system of medicine (2). India is endowed with Sanskrit and Indian bdellium in English (6). a rich genetic resource of medicinal plants and The genus Commiphora is widely distributed is rightly called the “Emporium of Medicinal in tropical regions of Africa, Madagascar, Plants” (3). Commiphora wightii (Arnott) is a Asia, Australia and the Pacific Islands (Good, medicinally important plant which is now 1974) (7). In India, it is found in arid, rocky considered as critically endangered species of tracts of Rajasthan and Gujarat Maharashtra 1896 Int.J.Curr.Microbiol.App.Sci (2021) 10(03): 1896-1902 and Karnataka (Kumar and Shankar, 1982) (20). It is not yet clear as to why plants die (8). In Rajasthan it is found in districts namely after tapping (21). Jaisalmer, Barmer, Jodhpur, Jalore, Sirohi, Ajmer, Sikar, Churu, Jhunjhunu, Pali, Seeds of guggul are the major propagation Udaipur, Alwar (Sariska Tiger Reserve), source in nature. In Rajasthan and nearby arid Jaipur (Ramgarh, Jhalana area), Bhilwara and regions flowers and seeds are constantly Rajsamand (9). Guggul is a woody shrub with produced by C. wightii except in winter knotty, crooked, sping brown bracties, leaves season. However, the germination of seeds are 1-3 foliate leaf lets, sessile with serrated poor thus large scale plantation is not possible margin (10). Fruit is drops red, ovate with two through this natural method. Therefore, celled stone (11). Flowers are small, brown, considerable efforts are still required to find pink flowers unisexual small, brownish red, out efficient in vitro methods for the the fascicles polygamous sexual distribution regeneration of this critically endangered bisexual, female and male flowers (12). Their medicinal plant. The in vitro propagation 3 - 4.5 mm long, usually red white pinkish, on method can be used for clonal propagation of the flowers individually or in groups appears selected germplasm, genetic improvement, in the 2 or 3 Fruits are red drupe, elliptical, production of active compound in cell culture. tapered - shaped, 2 - cell - type store, rarely In vitro propagation in C. wightii has been four your valve voice, when ripe will be red attempted through organogenesis and somatic and divides into two (13). The ash coloured embryogenesis methods by various bark comes off in flakes exposing the under researchers. bark which also peels off in thin papery tolls (14). The shrub defoliates in winter and Materials and Methods reserves for guggul gum extraction are high during April - May (15). This prominent The present investigation entitled species of the arid tracts of Rajasthan and “Micropropagation in Guggul (Commiphora Gujarat states (northwest India), Commiphora wightii (Arnott)) was carried out at the wightii is now on the verge of extinction over Department of Plant Breeding and Genetics, S. much of its Indian range and is listed as K. N. College of Agriculture, Jobner. The endangered (IUCN 2010) (16). The details of material and methods used in the predominant reasons for its fast diminishing present investigation are given below under populations are over - exploitation (tapping of separate headings: woody shoots for its oleo-gum - resin), poor natural germination rate and slow growth rate Plant Material (17). The resin extracted from stem is considered by some to have tremendous value The present research work was conducted on as cholesterol reducing agent and hence a Commiphora wightii. Nodal segments and favorite of the ayurvedic medicine industry shoot apexes were used as explants and (18). This has resulted in widespread obtained from healthy trees grown in S. K. N. indiscriminate tapping for the resin (19). The College of Agriculture, Jobner. magnitude of the conservation problem facing C. wightii through this exploitation is greatly Culture Medium exacerbated by the fact that a plant after being tapped through deep cuttings, usually dies All chemicals used in the present study were within two to six months of a single tapping of analytical grade. Murashige and Skoog episode (Bhatt et al., 1989, Paliwal, 2010) Medium (1962) was used throughout the 1897 Int.J.Curr.Microbiol.App.Sci (2021) 10(03): 1896-1902 course of investigation. After preparing the minutes with vigorous shaking. After washing stock solutions of various components, the with detergent, explants was again washed medium was prepared by mixing these stock with running tap water to remove any trace of solutions in a manner so as to maintain the detergent for 5 minutes. Finally explants were final concentration of each. Stock solution of surface sterilized with 0.1 per cent HgCl2 in a plant growth regulators were prepared by laminar air flow cabinet for 3 - 4 minutes. adding desired concentration of auxins and cytokinins. Culture conditions Glass ware All cultures were incubated at 25+20C under fluorescent light in a 14: 10 hour’s The borosilicate glasswares were used for all photoperiod. the experiments. Oven dried (2500C) Erlenmeyer flasks (conical flask), round Levels of antioxidants bottom flask, flat bottom flasks, pipettes, Petri dishes, beaker, measuring cylinders (50 ml, Antioxidants viz., activated charcoal and 100ml, 500 ml, 1000 ml and 2000 ml) and test polyvinylpyrollidone were used to control the tubes were used for media preparation. accumulation of phenolic compounds in the culture medium and enhance the rate of Autoclaving Micropropogation was worked out at most responsive level of plant growth regulators. Media were sterilized in autoclave. Distilled water, micronutrient and other stable mixtures Activated charcoal (20, 40, 60, 80 and 100 were autoclaved. The culture media contained mg/l) in glass containers sealed with cotton plugs and covered with aluminum foils were Polyvinylpyrrolidone (2, 4, 6, 8 and 10 mg/l). autoclaved at 15 psi. and 1210C for 15 - 40 minutes. Results and Discussion Sterilizing culture rooms and transfer area When different antioxidants incorporated singly in MS medium supplemented with Initially, the culture rooms were cleaned by responsive level (Nodal segment, 3.0 mg/l gently washing all the floors and walls with a BAP) of plant growth regulators for shoot detergent soap followed by daily cleansing multiplication elicited different response for with phenyls. Transfer area was sterilized by shoot bud induction because it controls the exposure to UV light. Aseptic condition of accumulation of inhibitory substances transfer area was maintained by installing an (phenolic compounds) in the growth medium. HEPA filter ventilation unit. Laminar airflow hoods were sterilized by wiping the working Effect of activated charcoal surface with 95 per cent ethyl alcohol. When activated charcoal (20 - 100 mg/l) was Explant preparation and sterilization added in the basal medium with responsive level of plant growth regulators (3.0 mg/l Explant was washed thoroughly in running tap BAP) in nodal segment, it induced shoots at water for 20 minutes, these were again washed all the level of activated charcoal ranging from with liquid detergent (RanKleen) for ten 1.37 – 1.55 within 16 – 18 days of incubation. 1898 Int.J.Curr.Microbiol.App.Sci (2021) 10(03): 1896-1902 Maximum number of shoot bud (1.55) was medium (Dumas and Monteuuis (1995), (b) observed at 60 mg/l activated charcoal with no adsorption of inhibitory substances, produced browning intensity in the culture medium. All by either the media or explants (Fridborg and other level showed low browning except 60, Eriksson, 1975, and Fridborg et al., 1978), (c) 80 and 100 mg/l AC (Table 1, Fig. 1). adsorption of plant growth regulators and other organic compounds (Constantin et al., Effect of polyvinylpyrrolidone 1977, and Weatherhead et al., 1978) and (d) the release of substances naturally present in Addition of polyvinylpyrrolidone in culture or adsorbed by activated charcoal (Johansson vessels with responsive level of plant growth et al., 1990). It was either beneficial or regulator induced shoot bud from nodal harmful effects on the culture, depending upon segment explants at all levels. Lower level of the medium, and tissue used under in vitro polyvinylpyrrolidone showed low browning in culture (Pan and Staden, 1999).
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