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A review on in vitro propagation of medicinal plants

Research · December 2018

DOI: 10.13140/RG.2.2.22551.42401

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 Journal of Pharmacognosy and Phytochemistry 2018; 7(6): 2228-2231

E-ISSN: 2278-4136
P-ISSN: 2349-8234
JPP 2018; 7(6): 2228-2231 A review on in vitro propagation of medicinal
Received: 19-09-2018
Accepted: 21-10-2018 plants
Naveen Gaurav
SGRR University, Patel Nagar Naveen Gaurav, Komal, Preeti Juyal, Manjusha Tyagi, Neha Chauhan
Dehradun, Uttarakhand, India and Arun Kumar
Komal
SGRR (PG) College Dehradun, Abstract
Uttarakhand, India Justicia adhatoda, usually known as malabar nut, Adulsa, Adhatoda, Vasa, Vasaka. In tribal name of
Justicia adhatoda known as KALA BASA (Encyclopedia, 2013). It is a medicinal plant native to Asia
Preeti Juyal widely used in ayurveda, homeopathy, unani and siddha medicines. Curing of diseases through medicinal
SGRR University, Patel Nagar plants is one of the oldest human practices and several traditional clinical procedures have been deputed
Dehradun, Uttarakhand, India using many plant species. From the last few decades, several hurdles related to the research for novel
drugs from natural products have been overcome through the use of new technologies, from
Manjusha Tyagi pharmaceutical approaches, including the integration of meta-bolomics and genomics approaches to
SGRR University, Patel Nagar
develop traditional methods of studying. The use of the plant for therapeutic purpose, whether in the
Dehradun, Uttarakhand, India
treatment or natural nutritional products. This has allowed the development of herbal medicines with
Neha Chauhan safety, efficacy and quality which stand out in the international pharmaceutical market. In the mid of
SGRR University, Patel Nagar 1981 and 2010, 34% of novel herbal drugs and medicines approved by US Food and Drugs
Dehradun, Uttarakhand, India Administration were based on molecules or direct derivatives from natural products, including statins,
anticancer, antimicrobial and immunosuppressant. Globally, the herbal medicines market trades
Arun Kumar spreading around $20 billion every year. The plant's range includes Sri-Lanka, Nepal, Bangladesh, India,
SGRR University, Patel Nagar Pakistan, Indonesia, Malaysia and China as well as Panama where it is thought to have been introduced.
Dehradun, Uttarakhand, India
Keywords: Propagation, medicinal plants

Introduction
The term tissue culture also referred to as cell grown in vitro or sterile culture media, has great
significance in both basic and applied studies. Primarily it is widely used in a wide sense to
include in vitro culture of plant cells, tissues as well as organs. Generally MS media is used for
tissue culture derived its name from the researcher Murashige and Skoog and their co-workers
(1962) (Gaurav, et al., 2015) [6]. In 1902, Haberlandt, was the first to invented cell culture and
for the idea of growing plant cells in an artificial media he is now famous as “father of plant
tissue culture”. Rapid advances in diverse aspect of plant tissue culture have been made during
the last few years and plant tissue culture techniques have been broadly used in agriculture and
industry. Plant tissue culture broadly refers to the in vitro cultivation of all plant parts, whether
a single cell, tissue or an organ under aseptic and optimal conditions (Ahmad, 2013) [2].
The underline principle involve in plant tissue culture in which a plant part from the mother
plant with an appropriate environment in which it can express its intrinsic or induced potential,
must be carried out aseptically. It has been found the plant can reproduce whole plant from
fragments of plant materials when given a MS media capable of supporting growth and
appropriate hormone control. This nutrient MS media used in plant tissue culture is an agar
media with macro and micro nutrients mixed in it. Unlike those plants which is growing from
a seed, tissue culture require a supply of carbon in an organic form such as sugars. They also
require amino acids, vitamins and growth hormones. The composition of the media will vary
with the plant material being cultured and some plants also successfully propagated in vitro in
B5 medium (Gaurav, et al., 2018) [8].
Plant tissue culture also used in research for geneticist, biochemist plant breeders and plant
pathologists and other researchers. Plant tissue culture has also proved more efficient in the
production of secondary metabolites and has been used in the commercial production of the
napthoquinones, pigments and shikonin. Plant tissue culture has also played a significant role
in the production of sweeteners, flavours, natural colorants and pharmaceuticals (Gaurav, et
al., 2016) [7].
Correspondence The term tissue culture was compose by American Pathologist Montrose Thomas Burrows.
Naveen Gaurav Tissue Culture is an important and very effective tool for the study of the biology of the cells
SGRR University, Patel Nagar from multicellular organisms. Plant tissue culture is a
Dehradun, Uttarakhand, India
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Journal of Pharmacognosy and Phytochemistry

laboratory technique abundantly refers to the in- vitro horticulture, the achievements and limitations of tissue culture
cultivation of the plant whether a single cell, tissue or an and some insights into current and possible future
artificial nutrient media under aseptic conditions. It is known developments. With rapid population growth, the total
as in vitro culture or micropropagation which generally acreages of fruits, vegetables and various ornamental plants
involves five distinct stages: have not been able to meet the needs of people in the
a) Selection of mother plant developing countries
b) Initiation of cultures K Tiwari, et al., (2000; 2010) [16, 17] A mass in vitro
c) Shoot multiplication propagation system devoid of growth regulators has been
d) Rooting of in vitro grown developed. MS medium supplemented with an antibiotic
e) Acclimatization (trimethoprim) or a fungicide (bavistin) supports direct shoot
bud induction on inter-node and leaf explants. Bavistin
It is based on the principle of tot potency of the cell. The use showed a marked cytokinin like activity thus optimum
of tissue culture technique was originated by German plant adventitious shoot buds induction occurred on MS medium
physiologist in an attempt to cultivated Mengkultarkan hair supplemented with 14μM bavistin. MS medium supplemented
cells of the mesophyll tissue of the leaf of monocot plant. with 0.44μM BAP and 1.14μM IAA (indole-3- acitic acid)
Plant tissue culture becomes a thrust area during the last few found most suitable for shoot elongation. In vitro derived
decades due to renewed emphasis it has received in all areas shoots exhibited better rooting response on MS medium
of crop improvement programmers (Shrivastava and Rajani, containing 4.9μM IBA. Regenerated rooted plantlets of
1999) [15]. Buddleja davidii franch were successfully acclimatized in
Tissue culture is mostly used in obtaining disease free plants, soil.
rapid propagation of plants those are difficult to propagate, Shashikanta Behera, et al., (2015) did work on an efficient
somatic hybridization, genetic-improvement of commercial plant regeneration protocol through two-stage culture process
plants, obtaining haploid plants from pollen cultures or of nodal segment has been reported for a significant medicinal
anther-culture for breeding programmes. plant, Buddleja davidii Franch. Multiple shoots with large
Techniques for cultivation of plant cells under defined number of shoot buds were induced from nodal explant on
conditions were learned through demonstration of tot potency Murashige and Skoog’s medium fortified with (1.0-5.0 mg L-
by R.J. Gautheret and P.R. White. Progressively novel 1) N6 Pennell through two-stage culture of nodal segments
techniques were soon available that leads to the application of and ex vitro acclimatization. Surface sterilization is most
tissue culture to five broad areas, namely:- important step before inoculation of explant. Different steps
1. Cell behavior (including cytology, nutrition, metabolism, have been employed for treatment of ex-plant for sterilization.
morphogenesis Scientists, have described sterilization treatment of B. davidii,
2. Pathogen-free plants and germplasm storage which includes use of 0.1% Mercuric chloride (w/v) for 2 min
3. Clonal propagation followed by rinsing thoroughly with distilled water. Different
4. Product (mainly secondary metabolite) formation, sterilization treatment was followed by Mathur and Kumar,
starting in the mid-1960s. Currently the various 1998, in which leaves and stem explants were shaken for 10
applications of genetic engineering are implemented in minutes in Tween-20 (Ranbaxy) and Savlon (Johnson &
medicinal plants to increase the production of secondary Johnson) in water for 10 minutes, rinsed in running water for
metabolites embryogenesis and pathology) 30 minutes, treated with 0.1% Mercuric chloride for 3-4
5. Plant modification and improvement,”.60s. minutes and washed several times with distilled water.
Tiwari et al. (2000) [16], suggested that for micropropagation
Akin-Idowu, P. E, Ibitoye, D. O and Ademoyegun, OT (2009) of Centella asiatica, plants were washed thoroughly for 30
[3]
. Over century ago, Haberlandt imagine the concept of plant minutes under running tap water followed by removal of
tissue culture and provided the ground work for the leaves, which was followed by soaking in the mixture of 1%
cultivation of plant cells, tissues and organs in culture. cetrimide solution containing 150 mg/l Bavistin (fungicide)
Initially plant tissue cultures arose as a research tool and and 50mg/l Trimethoprim for 25-30 minutes. The explants
focused on attempts to culture and study the development of were finally treated with 0.1% mercuric chloride for 3-4 minutes
small, isolated cells and segments of plant tissues. At the peak followed by rinsing in sterile distilled water for 4-5 times.
of the plant tissue culture era in the 1980s, in a relatively short Tiwari, et al., (1998) [16] did work on callus derived from
time, many commercial laboratories were established around nodal explants cultured on MS medium containing 0.5 mg/l
the world to capitalize on the potential of micropropagation 2,4-dichlorophenoxyacetic acid (2,4-D), when sub-cultured on
method for mass production of clonal plants for the MS medium containing 0.1 or 0.5 mg/l BAP or 0.2 mg/l 2,4-
horticulture industry. D + 0.1 or 0.5 mg/l kinetin, developed somatic embryos. The
Today plant tissue culture applications encompass much more somatic embryos germinated either on the MS basal medium,
in clonal propagation. The range of routine technologies has and the resulting plantlets were successfully transplanted to soil.
expanded to include somatic embryogenesis, somatic Rani, et al., (2003) [13] reported that callus induction in
hybridization, virus elimination as well as the application of Withania somnifera (L.) Dunal was observed from hypocotyl,
bioreactors to mass propagation of plants. Perhaps the greatest root, and Cotyledonary leaf segments, grown on Murashige
value of these tissue culture technologies lies not so much in and Skoog medium supplemented with various concentrations
their application to mass clonal propagation but rather in their and combinations of 2,4-dichlorophenoxyacetic acid (2,4-D)
role underpinning developments and applications in plant and kinetin (Kn). Maximum callusing (100%) was obtained
improvement, molecular biology and bio processing, as well from root and Cotyledonary leaf segments grown on MS
as being a basic research tool. Plant tissue culture technique medium supplemented with a combination of 2 mg/_M) 2,4-D
though an underutilized tool in Nigeria, it can be extensively and 0.2 mg/lM) Kn. When hypocotyl segments were used as
applied in horticulture to increase crop production. This paper explants, callus induction was noticed in 91% of cultures,
highlights some of the applications of plant tissue culture to which showed shoot regeneration on MS medium
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Journal of Pharmacognosy and Phytochemistry

supplemented with 2 mg /l, 2,4-D and 0.2 mg /l Kn. response but at higher concentration of the same hormones
A.H. A. Abdelmageed, et al., (2012) [1] did work on callus of shuts the callus growth. The concentration of BAP and 2, 4- D
M. champaca could be induced on MS media supplemented also shows good callus response in higher concentration
with 0.0 - 2.0 mg/l IAA, 0.0 - 2.0 mg/l 2,4-D, and 0.0 - 2.0 whereas low concentrations of these hormone combination
mg/l BAP, respectively. All growth regulators that were used shows nil effect. The morphology of callus differs upon the
in this study are capable of inducing callus from explants. hormonal concentration from green to white and green to
0.10 mg/l IAA and 0.10 mg/l 2, 4-D were found to be the brown with various textures. This protocol paves the way for
suitable concentrations for the induction of callus. The the development of in vitro regeneration for cotton and
survival rate of explants in the media supplemented with IAA consequently will promote the application of plant tissue
was found to be 81%. MS Media supplemented with different culture.
concentrations of BAP combined with IAA and 2, 4-D were
tested for callus proliferation. The callus growth showed Discussion
positive response to all growth regulators combinations tested Medicinal plants are potential renewable natural resources
in this study, except the combination BAP/IAA with 0.2/0.10 and are generally considered to play a beneficial role in
mg/I which exhibited low response with dark brown callus. human health care. The medicinal value of these plants
M. Priya Dharishini1, et al., (2014) did work on Callus and contains phytochemicals such as alkaloids tannins, saponins,
shoot induction was observed from leaf, node and internode flavonoids, and phenolic compounds as a secondary
explants of Buddlejadavidii Franch when grown in MS media metabolites. The most important is vasicine, a quinazoline
amended with varying concentrations of auxin and cytokinin. alkaloids are produced through PTC methods. In India, north
Concentration at 0.2 mg and 0.4 mg of 2,4-D showed 20 and part is under strategic geographical location and possesses an
33% of callus formation from leaf ex-plant. In vitro callus invaluable treasure of medicinal plants holding a major share
was induced from nodal explants in media amended with in cultivation and export.
napthyl acetic acid (NAA) (0.2 mg) and 6-benzyl amino
purine (BAP) (0.1 mg) wherein the growth response was References
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