Plant Cell Culture Techniques in Desmodium Gangeticum: a Mini Review

Sakshi Sharma1, Shivika Sharma2, Vikas Sharma1*

1*Author for correspondence: Dr Vikas Sharma, School of Bioengineering and Bioscience

LPU-Jalandhar

2 Dr Shivika Sharma, Asst Prof. DAV University, Jalandhar

Abstract

Plant based extraction of natural products demand is increasing with time. Plants belonging to the family of Fabaceae mainly desmodium species are most widely employed with desmodium gangeticum being most in demand. Plant cell culture techniques were used in this case for novel extraction of bioactive product from desmodium gangeticum. It was seen that desmodium gangeticum contain different types of products like pterocarpens, flavonoids etc. Gangetin was extracted and its phytochemical profile was enhanced. It was seen that it has an effect on fertility of females also act as an anti-inflammatory and analgesic.

Keywords : medicinal plants, desmodium gangeticum, invitro micropropagation

Introduction

Plant Biotechnology is involving a set of techniques used to adopt plants for specific needs or opportunities. Basically, it involves the usage of plants for the benefit of mankind. With the increasing problems faced due to chemical constituents used in foods and cosmetic industry it has been a great deal to find out some natural plant - based products. Natural plant -based products tend to be less toxic and have no adverse effects with their prolong usage.

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© 2019 JETIR January 2019, Volume 6, Issue 1 www.jetir.org (ISSN-2349-5162) The use of natural products for cosmetics, food and medicines have been in trend since long time. The use of plants for the extraction of natural products either in the form of pure compunds or extracts have always been useful. Currently the plant based extraction is being exploited for a manifold of applications. In the Public Health; to deal with many diseases natural extraction from plants can also prove beneficial. In India there are huge varieties of medicinal plants ranging from bael, tulsi, peppermint , desmodium etc. They vary on the basis of altitude variation and each part of plant can be used for extraction. Ranging from woody plant species we use bark whereas in the case of herbaceous species the use of bark, leaves etc can be taken for medicinal purposes. Like pinus roxburghii was found to be the most density tree in subtropical forest and rhobodendron arboretum was found in temperate region [ 1 ] .

Out of many varieties of medicinal plant present we had selected desmodium and its species .

Desmodium species are tend to be found all over ranging from sub tropical to tropical regions which include Asia, Afghanistan, India, Kashmir. It can never be formed in open grassy lands but in orchard weeds. In India it is grown commonly in waste lands, forests and plain areas.

Desmodium is the genus of the flowering plant family Fabaceae. It is regarded as inconspicuous legumes which have bright purplish flowers. Shalparni which is desmodium gangeticum is the most widely available plant which is of great use. It is 2 to 4 feet in height is a high erect undershrub with a woody stem and has branches covered with soft hairs. Seeds are compressed and are kidney bean shaped. It has leaves which are unfoliate, ovate and are upto 15 to 18 cm long. Fowering occurs mostly in two regions which is March to June and

September to October . Flowers are pink -violet in colour. The climate it requires for its growth is tropical and sub-tropical climate. It grow on loam to clay loam soil at approximately alkaline ph [ 2].

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© 2019 JETIR January 2019, Volume 6, Issue 1 www.jetir.org (ISSN-2349-5162) With the growing trend of natural products for cosmetics, food etc plant cell culture techniques use have been increasing in the last 10 years. Plant tissue culture is basically the collection of techniques used to maintain or grow plant cells, tissues or organs under sterile conditions on a nutrient culture. There are many different types of plant tissue culture techniques which are based on the explants used, these are callus culture ( which involves the culture of differentiated tissues from explant which differentitates in vitro to form callus). Organ culture is using embryo, seed, roots, endosperm etc. [3]. Cell culture is basically culturing of cells in vitro in nutrient medium which contains all macro and micronutrients, auxins and gibbrellins.

Mostly cells devoid of cell wall i.e. protoplast are being used. These plant cell culture techniques can be used for production of somatic embryo i.e. germplasm banks, for crop improvement, extraction of natural products etc. Plant cell culture techniques have been the most reliable and productive system for the production of bioactive products.

Demsodium species have exhibited a wide spectrum of pharmacological activities. Out of all the species desmodium gangeticum is one of the most useful species with enhanced profile so we have selected that. There are very huge varieties of compounds being isolated from desmodium these are like to contain alkaloids such as phenyl ethyalamines , pterocarpans and flavonoid glycosides like gangetin, desmodin, desmocarpin, o-beta-d-glycopyranoside etc. desmodium gangeticum is used as a tonic, febrifuge, anticatarrhal and anti-inflammatory etc.[4].

Gangetin is being regarded as a pterocarpenoid which was isolated from desmodium gangeticum. It tends to have a role in fertility of females, it was seen that the weight of sex organs could also be reduced after the gangetin treatment. It could also be checked to have anti-inflammatory, analgesic properties also. [5] Chromatographic techniques were used for identification and characterization of the properties of the compound present.

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The plant cell culture based extraction is based on the generation and establishment of a culture which could be generated by differentiated cells and dedifferentiated cells. Firstly, a suitable plant material is selected which is healthy and not suffering from any disease then sterilization, induction, callus generation etc. were performed.[6]. Surface sterilisation of plant material is needed to be done which is the most crucial step which is done by using mercuric chloride solution 2% or washing it with ethanol then repeated washing with distilled water.[7].

Each plant part could contain the properties but only that particular plant or organ type is taken which is based on the biochemical product extraction which we want to do. Growth regulators like phytohormones, auxins also are taken into consideration. Roots of desmodium gangeticum are taken as these are highly productive for the extraction of natural product. Selection of optimum plant material is needed because the production of natural products cannot occur if there are some cells which have loss their capacity to dedifferentiate.[8] For the production of natural products the most important requirement after the selection of plant material is the culture condition optimization and establishment. There are some essential plant cell culture mediums these are MS medium (Murashige and skoog medium), white ‘s medium and gamborg medium. But the most used medium is MS medium, but the growth and extraction can be enhanced by providing the medium with good sources of energy, growth hormones, nitrogen and phosphate regulators etc. [9].

It was seen that the production of plant cell growth and natural products were inversely related.

So we can say that the production of natural products would occur as a source of enzymes in secondary metabolism phase of growth. So we have to transfer the plant material into cell culture growth medium into a primary production media which basically involves the reduction

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As for the extraction of product from desmodium we culture the explants into the MS mesium which could be supplemented with either IBA or IAA [10].

Elicitation of plant based natural production could also occur in response to stress and elicitation thus that can also be induced by plant pathogen. It was seen that gangetin that was present in the roots of desmodium gangeticum was seen to be enhanced in the presence of auxin that was seen when supplementing the effect of gangetin in rats. [11]

Permeabilization and extraction of natural products

Extraction and permeabilization of natural products is important, permeabilization involves the removal or extraction of natural product without disturbing the growth and viability of cells. This technique basically involves removal of natural product either with treatment with solvent like dimethylsufoxide and isopropanol. The most crucial step in the production of natural products is there extraction, this include pre-washing, drying, freeze-drying, homogenisation step. The extraction can be done on the basis of liquid liquid system, insitu filteration etc.[12]. Another technique that could be employed was known as biotransformation which could be used in production of chemicals in plant based extraction. Cambial meristematic cells can be used as culturing and application of natural products. Scale up for the production of natural products can also be done. These large scale culturing of plant can be done in bioreactors that is stirred tank, column bioreactor.

Identification and characterization of these natural products can be done by chromatographic techniques like TLC, HPLC and Gas chromatography these can be used to identify pure compound and biochemical activity and characterization of this pure compound and structure can be determined by this. Desmodium gangeticum tends to contain huge amount of compounds TLC can be used to determine the identity of a compound in a mixture by JETIRDW06096 Journal of Emerging Technologies and Innovative Research (JETIR) www.jetir.org 619

© 2019 JETIR January 2019, Volume 6, Issue 1 www.jetir.org (ISSN-2349-5162) determining the retention factor [13]. Gangetin was a pterocarpenoid isolated from the roots of desmodium gangeticum, after its phytochemical profile was determined it was seen that it has an effect on the fertility of females, it was determined that it could have an effect on 87.50

% antiimplantation effect on female rats. The weight of the sex organs could also be decreased after the gangetin treatment [14]. It was seen that it could suppressed the fertility. Also it was seen that gangetin has specific properties like analgesic, anti-inflammatory etc.

Conclusion

Here we had documented the use of plant cell culture techniques for the extraction of natural products from Desmodium gangeticum which is a potential source of extraction of natural products. Thus plants have evolved the capacity for the production of natural, bioactive products which has more benefit as compared to the chemical. The use of plant cell culture techniques has increased the capacity of manufacturing of natural products with less energy utilization and easy process. It was seen that natural products since a long time have been in demand and thus its use have been increased and thus production is increasing in demand.

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© 2019 JETIR January 2019, Volume 6, Issue 1 www.jetir.org (ISSN-2349-5162) 5. K.K. Purushothaman, V.M. Kishore, V. Narayanaswami, J.D.N. Connolly, “ The structure and stereochemistry of gangetin a new ptero-carpon from Desmodium gangeticum (Leguminaceae),” J. Chem. Soc, pp. 2420-2422, 1971.

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