Plants in Vitro in Food & INDUSTRI

Plants in vitro propagation with its applications in food, pharmaceuticals
and cosmetic industries; current scenario and future approaches

Ammarah Hasnain, 1 , * Syed Atif Hasan Naqvi, 2 , * Syeda Iqra Ayesha, 1 Fatima Khalid, 1 Manahil
Ellahi, 1Shehzad Iqbal, 3 Muhammad Zeeshan Hassan, 2 Aqleem Abbas, 4 Robert Adamski, 5 Dorota
Markowska, 5 , *Alaa Baazeem, 6 Ghulam Mustafa, 7 Mahmoud Moustafa, 8 , 9 Mohamed E.
Hasan, 10 and Mohamed M. A. Abdelhamid 11
Author information Article notes Copyright and License information PMC Disclaimer
This article has been corrected. See Front Plant Sci. 2023 April 17; 14: 1197747.
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Abstract
Plant tissue culture technique employed for the identification and isolation of bioactive
phytocompounds has numerous industrial applications. It provides potential benefits for
different industries which include food, pharmaceutical and cosmetics. Various agronomic crops
i.e., cereals, fruits, vegetables, ornamental plants and forest trees are currently being used
for in vitro propagation. Plant tissue culture coupled with biotechnological approaches leads
towards sustainable agricultural development providing solutions to major food security issues.
Plants are the rich source of phytochemicals with medicinal properties rendering them useful
for the industrial production of pharmaceuticals and nutraceuticals. Furthermore, there are
numerous plant compounds with application in the cosmetics industry. In addition to having
moisturizing, anti‐ageing, anti‐wrinkle effects; plant-derived compounds also possess
pharmacological properties such as antiviral, antimicrobial, antifungal, anticancer, antioxidant,
anti-inflammatory, and anti-allergy characteristics. The in vitro propagation of industrially
significant flora is gaining attention because of its several advantages over conventional plant
propagation methods. One of the major advantages of this technique is the quick availability of
food throughout the year, irrespective of the growing season, thus opening new opportunities
to the producers and farmers. The sterile or endangered flora can also be conserved by plant
micro propagation methods. Hence, plant tissue culture is an extremely efficient and cost-
effective technique for biosynthetic studies and bio-production, biotransformation, or
bioconversion of plant-derived compounds. However, there are certain limitations of in-
vitro plant regeneration system including difficulties with continuous operation, product
removal, and aseptic conditions. For sustainable industrial applications of in-vitro regenerated
plants on a large scale, these constraints need to be addressed in future studies.
Keywords: plant tissue culture, explants, secondary metabolites, industry, pharmaceuticals,
medicines, cosmetics
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1 Introduction
Plant tissue culture is a technique in which fragments of tissues from a plant (explants) are
developed in vitro in an artificial medium under aseptic conditions. It involves culturing explants
(such as shoot tip, root tip, callus, seed, embryo, pollen grain, ovule or even a single cell)
isolated from mother plant on asterile nutrient medium which leads to cell multiplication and
plant regeneration (Ravishankar and Venkataraman, 1990; Veeresham and Chitti, 2013; Kaya
and Huyop, 2020; Vidyagina et al., 2021). The commonly used medium in plant tissue culture is
Murashigeand Skoog (MS) basal medium (Murashige and Skoog, 1962) supplemented with the
required amounts of plant hormones which include auxins, cytokinins, abscisic acid,
gibberellins, ethylene, and growth regulators with similar metabolic effects (Gaspar et al.,
1996; Arab et al., 2014; FargosoMonfort et al., 2018; Kaya and Karakütük, 2018).
The first reports on tissue culture came from the early twentieth century when Gottlieb
Haberlandt (Haberlandt, 1902) undertook experiments to sustain mesophyll cells in culture
based on postulates establishing the “toti potentiality of plant cells.” Since then, progress has
been steady, with hundreds of results and publications on the use of tissue culture techniques
in breeding programmes, genetic biodiversity conservation, and biopharmaceutical
manufacture being reported each year (García-Gonzáles et al., 2010). Initially, plant tissue
culture was utilized as a research tool to study the development of isolated fragments of plant
cells and tissues; however, the advent of recent molecular biology techniques significantly
broadened the avenue of experimental investigations and applications (Chen et al., 2016; Tegen
and Mohammed, 2016).
Plant tissue culture plays a significant role in basic research in the areas of plant pathology, plant
physiology, plant metabolites and conservation (Espinosa-Leal et al., 2015; Chandran et al.,
2020; Vidyagina et al., 2021; Taalat et al., 2021). Plant micro-propagation has extensively been
used to have an insight into plant pathology studies which include factors influencing
penetration, infection and multiplication of pathogens, the nature of irregular cell division or
growth, and the morphogenetic potential of the diseased cell (Maheshwari, 1969; Köhl et al.,
2019). The employment of plant tissue culture in plant pathology is not only restricted to use
plant tissues as a substrate for the pathogens, but it provides the basic understanding of various
characteristics of pathological growth, pathogens’ attack weapons, and the host response to an
infection caused by the invading organism (Maheshwari, 1969). Plant physiology and plant
morphogenesis requires the capability to grow plants in vitro that might be best accomplished
with plant tissue culture procedures (Pullaiah and Subba, 2009). Furthermore, the conservation
of plant biodiversity is indispensable for future crops safety due to increasing challenges of
biotic and abiotic stresses (Alexandratos and Bruinsma, 2012; Gupta et al., 2016). In this
regard, in vitro techniques permit improvement in various traits associated with plant growth
and yield that can, later, be used for ex-situ conservation (Lavanya et al., 2014). The active plant
compounds obtained from rare or endangered species can be manufactured by in
vitro techniques without adverse environmental effects and in agreement with the bio-
sustainability matters that the market demands (Figures 1, 2).
Figure 1
An overview of tissue culture process (A, B) small explant develops callus which then produces
shoots a few weeks after being placed into tissue culture media (C) “A to I” shows complete
procedure from single cell placement to MS media to development of a complete plant (D) How
all phases in plant tissue culture from initiation, multiplication, root formation, shoot formation
and acclimatization occurs.
Figure 2
In vitro plant propagation of plants at Tissue Culture Lab (A, B) Roots are fully developed prior
to moving plants to pots of soil.
The in vitro plant propagation has not only made a significant contribution in the knowledge of
basic research, but it also offers potential applications as it guarantees a sustainable industry
that relies on commercial production of plant-derived compounds (Kumari et al.,
2021; Chandran et al., 2020). The culture of plant tissues is an effective instrument for the
isolation and processing of active compounds, including secondary substances and engineered
molecules, from economically important plants (Espinosa-Leal et al., 2018; Kumari et al., 2021).
Due to advancement in contemporary techniques, several protocols have been developed for
the production of a wide variety of plants secondary metabolites on a commercial scale
(Shasmita and Naik, 2018). Furthermore, plant tissue culture coupled with biotechnological
approaches is applicable to the development of genetically modified plants as well as embryo
rescue procedures (Altpeter et al., 2016; Tegen and Mohammed, 2016; Sadiku et al., 2018). It
plays a pivotal role in vector-mediated or vector-independent gene-delivery into plant genome
for the production of transgenic plants with improved traits (Hussain et al., 2012; Gleba et al.,
2014; Hasnain et al., 2020).
Various crops with superior traits have been developed using this technology with enhanced
nutritional value and biotic/abiotic stress resistance that leads to increased crop yield
(Ravishankar and Venkataraman, 1990; Rishirumuhirwa, 2007; Ragavendran and Natarajan,
2017). Different transcription factors which regulate nutrient assimilation pathways have been
over expressed in staple crops that may improve crop yield (Kurai et al., 2011; Hasnain et al.,
2020). Plant propagation using tissue culture techniques have also provides a valuable
commercial prospect in the industrial manufacture of ornamental plants, vegetable and fruit
plants with economically important products (Hussain et al., 2012). The current review gives
insights into the research that has been conducted successfully on in-vitro plant regeneration in
order to obtain valuable bioactive phytocompounds having several applications. The review also
focuses on major constraints associated with the use of plant tissue culture technique that need
to be addressed for sustainable industrial applications. Various useful products can be obtained
as food products, pharmaceutical products and cosmetics via in vitro plant regeneration
(Niazian et al., 2017; Chandran et al., 2020). However, plant tissue culture studies are needed to
be conducted on a large scale in order to ensure food security and meet the demand of food
supply of increasing human population (Ragavendran and Natarajan, 2017). In this review we
have discussed micro-propagation of some plants species to have a better understanding of
significance of pant micro-propagation in different industries.
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2 Plant propagation types
The two types of plant propagation are mentioned as follows
2.1 Macro-propagation
Macro-propagation is a method used in a shed or in the field. It entails producing suckers from
clean planting material by eliminating apical dominance. Macro-propagation techniques are
divided into two types: field-based techniques that rely on total or partial decapitation, and
detached corm techniques that are done in a shed. Traditional methods of offset planting,
rhizome planting, cutting roots, and layering are used in macro-propagation (Njukwe et al.,
2007).
2.2 Micro-propogation
Plant micro propagation, also known as plant tissue culture, is a technique that isolates,
sterilizes, and incubates cells, tissues, or organs of chosen plants in a growth-promoting aseptic
environment to create a large number of plantlets. The isolated cloning technique revealed that,
given the right conditions, somatic cells may develop into a complete plant (Njukwe et al.,
2007).
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3 Plant tissue culture methods
Several methods are available for plant tissue culture. In organogenesis, the commonly used
method,organ formation can occur directly from meristems, or indirectly from dedifferentiated
cells (callus). The resultant cultures can then be utilized to mass produce plants (micro
propagation) or to develop specific organs (e.g., roots in hairy root culture) (Espinosa-Leal et al.,
2018).
3.1 Organogenesis
Organogenesis is the production of plant organs from a specific tissue in order to develop
complete plants. It is characterized by being polar, which means that just one aerial organ or
root is released and a new complete plant is generated from this. Simultaneously,
organogenesis can be direct in which the organogenic shoot is produced directly from the
explants, or indirect, in which the organogenic process happens from previously created callus
in the original explants (Gupta et al., 2020).
3.2 Somatic embryogenesis
Somatic embryogenesis is the process of producing embryos from somatic plant cells (any non-
sexual cell) in order to produce a whole plant. In contrast to organogenesis, this is a polar
process in which the aerial structures and roots of plants develop from the somatic embryo. It
can either be direct or indirect, depending on whether the process begins with the original
explants or with previously produced callus (Gupta et al., 2020).
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4 Types of tissue culture
4.1 Callus culture
Callus is an undifferentiated mass of tissue that forms on explants after a few weeks on growth
medium with appropriate hormones. Callus development is the result of a well-known process
known as de-differentiation or re-differentiation. To stimulate callus induction and
development, several growth hormones are employed. Callus induction and development were
increased in Cephaelis ipecacuanha by 2,4-D, NAA, and kinetin. Organogenesis allows for the
effective regeneration of new plants from callus (Ujjwala, 2007; Espinosa-Leal et al., 2018).
4.2 Suspension culture
In vitro suspension cultures are created when friable calli are grown on liquid medium in a
suitable container and regularly agitated to provide free cell suspension. Conical flasks are
utilized because of its enormous surface area, which aids in the retention of liquid medium and
the constant exchange of gases. Suspension cultures are classified as batch or continuous
cultures. At regular intervals, a part of the original cell suspension is collected and sub-cultured
on to fresh medium in batch cultures. In continuous cultures, new media is introduced to the
same culture on regular basis, and surplus cell suspensions are discarded. Suspension cultures
are commonly utilized in large-scale synthesis of secondary metabolites. Chemostat bioreactors
are the devices particularly built for large-scale continuous culturing (Smith and Drew,
1990a; Smith and Drew, 1990b; Baezas-Lopez, 1995; Kodym and Zapata-Arias, 2001; Ahloowalia
et al., 2004; Bimonte et al., 2011; Sidhu, 2011; Chen et al., 2015; Chen et al., 2016a; Titos et al.,
2016; Suman, 2017).
4.3 Meristem culture
The culture in which tiny excised shoot apices are cultivated, each having an apical meristemetic
dome with one or two leaf primordia. Typically, the shoot apex is developed to produce a single
shoot (Ahluwalia et al., 2016).
4.4 Protoplast culture
Protoplasts are plant cells with cell walls removed by enzymatic or mechanical methods.
Protoplasts are obtained by immersing plant cells in a hypertonic solution, which causes the
plasma membrane to shrink off the cell walldue to water efflux. Cell wall may now be removed
using either enzymatic digestion with pectinase and cellulose, or by mechanical techniques
(Sidhu, 2011).
4.5 Shoot tip culture
Shoot tip culture is developed from excised shoot tips/buds larger than the shoot apices (used
for meristems cultures), and had several leaf primordia. These shoot apices are often cultivated
such that each one generates several shoots (Espinosa-Leal et al., 2018).
4.6 Lateral bud node culture
Lateral bud node culture is carried out on a short piece of stem tissue where stem portions
carrying single or many nodes may be cultivated. Each bud is cultivated to produce a single
shoot (Sidhu, 2011).
4.7 Isolated root culture
In isolated root culture, a branching root system can be generated by growing roots that are not
attached to shoots (Ahluwalia et al., 2016).
4.8 Embryo culture
Embryo culture has fertilized or unfertilized zygotic (seed) embryos dissected from maturing
seeds or fruits and cultivated in vitro until seedling formation. Embryo culture is not the same as
somatic embryogenesis (Gupta et al., 2020).
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5 Advantages of in vitro plant propagation
Plant micro-propagation with different explants like seeds, embryos, calli, anthers, protoplasts,
and meristemetic tissues of root/shoot tips is used for large-scale production of industrial
products (Mali and Chavan, 2016; Shasmita and Naik, 2018). Conventionally, somatic
hybridization, which produces interspecific and intergeneric hybrids, was commonly used as an
essential method for plant breeding. The procedure entails the fusion of two somatic
protoplasts followed by the selection of desirable hybrid cells and then regeneration of hybrid
plants (Lambardi et al., 2008). Protoplast fusion is an effective method of transferring genes
with desirable traits from one species to another with a great impact on crop development.
Somatic hybrids created from rice and ditch reed by electrofusion exhibited better results
against salt stress (Hussain et al., 2012). The most recent feature of plant cell and tissue culture
is genetic transformation, which allows for the transfer of genes with desirable traits into host
plants and the recovery of transgenic plants (Hinchee et al., 1994). This approach offers a high
potential for the development of agricultural plants with novel traits. The genetically modified
plants exhibit agronomically significant features such as greater yield, improved nutritional
quality, improved pest and disease resistance (Sinclair et al., 2004).
Recent breakthroughs in plant cell culture, molecular biology, enzymology, and fermentation
technology indicate that these systems are a viable source of synthesis ofimportant secondary
metabolites. Plants infected with an engineered virus generate relatively significant amounts of
desired chemicals, and these plants can sustain steady levels of protein synthesis without extra
intervention (Sajc et al., 2000). Large-scale plant tissue culture has been shown to be an
appealing alternative approach to traditional plantation methods since it provides a regulated
supply of biochemicals independent of plant availability (Sajc et al., 2000). Kieran et al.
(1997) examined the effects of several engineering parameters on cell suspension cultures for
the recovery of important metabolites (Vanisree et al., 2004). Tissue culture technology
advancements show that transcription factors are effective new molecular tools for plant
metabolic engineering to boost the synthesis of important chemicals (Gantet and Memelink,
2002; Hasnain et al., 2020). In vitro cell culture has the inherent advantage of producing
therapeutic proteins such as monoclonal antibodies, antigenic proteins that act as immunogens,
human serum albumin, interferon, immuno-contraceptive proteins, antihypertensive drug
angiotensins, and human haemoglobin in certain situations (Wongsamuth and Doran, 1997).
Tissue culture technique offers several advantages over plant propagation under natural
conditions (Bhojwani and Dantu, 2013; Chandran et al., 2020). It is a rapid procedure as
thousands of seedlings can be produced from small fragments of plants in a short period of time
in contrast to conventionally propagated flora (Mattick, 2018). This also helps to accelerate the
production process of new crop varieties with superior traits as tissue culture experiments
require less time and space compared to in-vivo plant growth (Krasteva et al., 2020). Tissue
culture can be used to propagate perennial plants; irrespective of weather or season (Ahmad
and Anis, 2007). It also helps in the development of pathogen-free micro-plants saved from
various diseases and the new plants produced by tissue culture under aseptic conditions are
also sterile (Chiipanthenga et al., 2012; Bayoudh et al., 2015; Tegen and Mohammed, 2016).
Furthermore, several plant species generate resistant seeds that cannot be retained for
extended periods of time; in this situation, tissue culture can be utilized for plant conservation
in vegetative state, generally under slow growth conditions (Lambardi et al., 2008), or for
cryopreservation (García-Gonzáles et al., 2010). In some cases, inter-specific and inter-generic
hybrids can be obtained using embryo rescue technique which is not possible through
conventional methods (Mattick, 2018). Tissue culture has been extensively utilized in breeding
programs for over 50 years. The hybrids of such crosses are often sterile due to embryo abortion
but can be ‘rescued’ by means of culturing or transplanting the embryos. The media conditions
used for the cell culture can be modified accordingly to provide more acceptable results for the
performed experiments (Pant, 2014).
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6 Applications of plant tissue culture
Since earliest times, humanity has been dependent on plants for food, flavors, medicines and
many other uses (Rai et al., 2011). Because the spectrum of phytochemicals is larger than that
of any other class of creature, plants are the most plentiful source of herbal remedies in the
food and cosmetics sectors (Barbulova et al., 2014). Plant tissue culture can be used for a wide
range of purposes with various applications in research and industry (Rai et al., 2011; Niazian
et al., 2017; Chandran et al., 2020). The major commercial applications which in vitro plant
propagation offers are in the following major industries (Table 1).
Table 1
Applications of plant tissue culture technique in different industries.
Industry Products Applications Limitations References
and
advantages
Food Rice Gives high Produce (Roy et al.,
yield less 2000; Sparks
irrespective of resilient and Jones,
seasonal plants, Less 2014; Yaqoob
requirements acceptance et al.,
and require towards 2016; Hasnai
less area for tissue n et al.,
production cultured 2018; Hasnai
Wheat Wheat food plants n et al., 2020)
varieties are in general
resistant public, no
against need of
different herbicide
diseases with and
increased yield pesticide
and high application
nutritional
value
Pineapple (Smooth Cayenne) High yield,
Gives efficient
and fast
production of
required flora
Banana Cheap and
sterile planting
materials
ultimately
giving a high
yield during
whole year
Chocolate Unique taste
and aroma
Pharmaceutica Codeine To relieve pain Less (Czyz et al.,
l as well as production 2014; Hayden
and
advantages
coughing of et al.,
Atropine Treat the secondary 2014; Mali
symptoms of metabolites and Chavan,
decreased , difficult to 2016)
heart rate remove
secondary
Reserpine Treat high metabolites
blood pressure from the
Hyoscyamine Treat a range culture
of
gastrointestina
l disorders
Digoxin Used to treat
heart failure
Scopolamine Avoid nausea
and vomiting
induced by
motion
sickness
Morphine Pain killer
Cosmetics Rubusidaeushydrosoluble Anti- It is difficult (Ebile et
extract inflammatory to maintain al.,2018)
activity in skin aseptic
cells conditions
Nicotianasylvestriscell wall Collagen resulting
preparation synthesis and high cost.
protection in
skin cells
Coffeabengalensishydrosolubl Epidermal
e extract hydration and
collagen
synthesis in
skin cells
Dolichosbiflorushydrosoluble Anti-
extract inflammatory
and
advantages
Malusdomesticus whole activity and
lysate UV damage
protection
Reversion of
aging signs
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6.1 Food industry
Plants are main source of life on planet earth for living creatures and exclusively providing
nutrition to humans and being the source of phyto-constituents like starches, proteins, dietary
fibers, minerals and cancer preventive agents (Khosroshahi et al., 2006; Nasr et al., 2007). Plant
tissue culture is a powerful tool of agricultural improvement and offers tangible solutions to
major crop problems that arise due to constant threat of biotic and abiotic stresses minimizing
the crop yield (Ragavendran and Natarajan, 2017; Eibl et al., 2018). Plant tissue culture coupled
with different biotechnological approaches leads to a sustainable agricultural ensuring food
productivity and safety (Alexandratos and Bruinsma, 2012; Bhojwani and Dantu, 2013). Various
transgenic plants including Arabidopsis, wheat and tobacco are developed through genetic
engineering and plant tissue culture conferring resistance against different environmental
stresses (Voytas and Gao, 2014; Hasnain et al., 2020). Since plant tissue culture is simple, low-
cost and environment friendly, it is imperative to employ this technique for the development of
sustainable agriculture in order to meet the food demand of increasing human population
(Gahakwa et al., 2012; Hussain et al., 2012; Altpeter et al., 2016). The greatest value of plant
cell/tissue culture rests not so much on their application to mass clonal propagation (micro
propagation), but also in their involvement in plant improvement and bio-processing. Its uses
extend well beyond crop cultivation and productionas it has an enormous potential of bridging
the gap between the research institutes and industry. The agro-industry based projects are
crucial to achieve agricultural sustainability enabling the ultimate target, the stakeholder, reap
benefit of extensive research being done across the globe (Suman, 2017). In this review we have
discussed micro-propagation of some fruits and crops with high nutritional value to have a
better understanding of role of plant tissue culture towards sustainable agriculture (Figure 3).
Figure 3
Recent methods used for industrial production of bioactive compounds via plant tissue culture.
6.1.1 Banana tissue culture
Common bananas are cultivars of Musa acuminate, native to South Asia. They are one of the
essential fruits that are being consumed as a staple food in the developing countries (Njuguna
et al., 2003; Dubois et al., 2013). Banana is also regarded as an important export produce in
tropic regions of the world; however, its production has been reduced during the last few years
(Macharia et al., 2010; Liu et al., 2019). Because banana is an essential food crop and the
second most significant fruit crop after mango, considerable care has been given to recording
the stages needed in effective micro propagation of banana. Despite the banana crop’s high
nutritional and economic value, the major production restriction is due to lack of dependable
and safe planting material. Diseases, pests, and a lack of planting materials have all been
blamed for the drop in production. Planting materials obtained by traditional means (suckers)
are insufficient to fulfill the growing demand for planting and are of low quality. Tissue culturing
is an efficient method for addressing problems pertaining to its low production (Strosse et al.,
2004). The challenge involved with conventional banana breeding is better understood by the
fact that no new commercially suitable banana cultivar has been generated over 60 years of
continuous breeding efforts (Rowe, 1984). In fact, bananas are one of the few crops that are
grown using exclusively clones produced from natural somatic mutations.Each mother plant
produces one to eight suckers in line, which is insufficient (Ko et al., 2009; Rossmann et al.,
2012). The solution is to propagate cheap and sterile planting materials using tissue culture
technique whilethe micro propagation of bananas will not only lead to fast multiplication of
desirable banana cultivars butproduces disease-free planting material may also be made
available throughout the year (Dubois et al., 2013; Liu et al., 2019; Selvakumar and Parasurama,
2020).
Agricultural diversification to satisfy our future demands necessitates the implementation of
innovative agricultural technology. The finest cultural methods, excessive fertilizers, and pest
control procedures will not yield the desired results unless the best planting material is used.
Tissue culture is now widely being used as a viable horticultural propagation technology, and it
has changed the horticultural business. This approach is used to achieve mass proliferation and
the creation of disease-free stock material (Suman, 2017).
Various studies depict that micro propagated bananas are capable of outperforming compared
to conventional planting material techniques (Smith and Drew, 1990a; Dubois et al.,
2013; Suman, 2017). However, the prevalence of somaclonal variations, referred to as off-types
in the industry, remained the most serious difficulty that developed to limit widespread
acceptance of micropropagated banana planting material. Plantings from commercial
laboratories have been found to include up to 90% off-types (Walduck et al., 1988), with
dwarfism being the most prevalent off-type detected. Dwarfs are more prone to ‘choke-throat,’
a physiological condition in which the bunch fails to emerge entirely from the plant, and their
hands are more densely packed, making dehanding more difficult (Smith, 1988).These
difficulties can be overcome by altering the type and concentrations of phytohormones in
culture media, the number of subcultures, and selecting different types of explantse.g. axillary
buds are more stable than adventitious buds. One of the most important intrinsic determinants
is the genetic stability of the cultivar or genotype. While there is little that can be done to
control intrinsic variables, micro propagation, is comparatively useful in modulating the intrinsic
factors by controlling external factors that would be, otherwise, difficult under natural
conditions (Walduck et al., 1988; Hasnain et al., 2020). Commercial laboratories are now using
tissue culture protocols that minimize somaclonal variations, and working on creating accurate
screening and selection approaches for early detection of off-types.Although somaclonal
variation is deleterious to quick clonal multiplication, some off-types have been discovered that
have significant agronomic utility. ‘Mons Mari,’ a micropropagated Cavendish cultivar, with
extra-long finger length for all hands and fruit (2-3 cm) longer than typicalis one of the
examples. This selection has the potential to increase profits through extra-large fruit sales. A
dwarf form with no visible choke-throat difficulties, larger, more open hands, and may exhibit
higher wind resistance than ‘Williams’ isanother option. These plants are now being replicated
for field testing in a variety of Queensland locations (Walduck et al., 1988; Bestoso et al.,
2006; Chen et al., 2016b; Reinhardt et al., 2018) (Figure 4).
Figure 4
In vitro propagation of banana (A) Callus formation to roots development (B) Maturation of
plants in media (C) Plant sown in pot (D) Plant sown in soil in controlled conditions,
acclimatization (E) Sowing of plant in the field (F) Tissue culture produced Banana field.
6.1.2 Pineapple tissue culture
In tropical places, the pineapple is a popular vegetatively propagated crop; nevertheless,
pineapple production is limited due to a lack of planting materials (Be and Debergh,
2006; Nikumbhe et al., 2013). Suckers and slips were traditionally used to propagate pineapple,
resulting in a limited number of seedlings that did not meet farmer demand. Tissue culture
provides a practical technique to produce the needed amount of desired flora in this regard
(Roy et al., 2000). Explants produced from crown-tip meristems are sterilized and cultivated on
MS media enriched with cytokinins in an efficient and cost-effective procedure for commercial
micro-propagation of Smooth Cayenne (a spine-free variety of pineapple). Direct organogenesis
is used to regenerate several shoots, which are then rooted in auxins-supplemented medium.
After that, the rooted shoots are planted in a sterilized soil substrate (Usman et al., 2013; Akin-
Idowu et al., 2014; Reinhardt et al., 2018). In other cases, auxiliary buds are taken from slips or
suckers and put on different medium to promote explant establishment followed by single
shoot growth, shoot proliferation, and finally root initiation. Preliminary studies onin
vitro propagation of pineapple by Geoge and Sherrington (1984) demonstrated that
multiplication rates of 30 to 50 per month were obtained on medium containing
benzylaminopurine (BAP), compared to 4-5 per year with traditional propagation. The use of
large quantities of cytokinins in multiplication media, on the other hand, has been linked to the
induction of somaclonal variations (Geoge and Sherrington, 1984). Plantings from commercial
laboratories with a multiplication rate of four per month contained a maximum of 5% variations
(predominantly rough-leaved types) from a total of ten thousand plants. This indicates an
appropriate amountgiven the benefits of nursery establishment.Moreover, pineapple plants in
their first generation of in vitro propagation have shown a high number of slips and suckers. This
has been beneficial for increasing planting material in nursery blocks (Figure 5).
Figure 5
In vitro propagation of pineapple (A, B) Initiation and multiplication stage (C–E) Root and shoot
formation, plants are fully grown to move to pots of soil.
6.1.3 Wheat tissue culture
Wheat (Triticum aestivum L.) belonging to family poaceae is one of the most important staple
crops worldwide as it plays a crucial role in meeting human nutritional requirements (Rashid
et al., 2012; Contardo-Jara et al., 2018). To fulfill daily nutritional requirements of growing
human population, wheat production should be increased to an annual rate of 2% (Sparks et al.,
2014). Wheat tissue culture is one of the alternatives of conventional breeding strategies that
may lead to the development of wheat varieties resistant against different diseases (Sparks and
Jones, 2014; Hasnain et al., 2018; Hasnain et al., 2020). The transgenic wheat lines conferring
resistance against take-all disease which is damaging to plant roots (Liu et al., 2013) and sharp
eyespot (Chen et al., 2008) have been developed using tissue culture and biotechnological
approaches. Callus culture is widely being used to develop plants conferring resistance against
various diseases responsible for drop in crop yield (Efferth, 2019). Over the last few decades,
one of the main research objectives is to enhance wheat production in order to meet the
demands of increasing human population. The use of different biotechnological approaches is
quite promising in attaining these objectives (Contardo-Jara et al., 2018; Hasnain et al., 2018).
The transcription factors involved in carbon and nitrogen metabolism pathways are over
expressed in wheat varieties, using tissue culture technique, which proved to be promising in
increasing wheat yield (Pena et al., 2017; Hasnain et al., 2020) (Figure 6).
Figure 6
Wheat tissue culture (A) Inoculation of single cell to tissue culture media (B) Root and shoot
development (C, D) Roots are fully developed prior to moving plants to pots of soil.
6.1.4 Rice tissue culture
Rice (Oryza sativa L.) has been cultivated for more than 7000 years as a major food crop and
presently more than 50% of world’s human population is dependent on it (Afolabi et al.,
2008; Ahmad et al., 2013). Rice is a rich source of nutritional carbohydrates supplying 50-80% of
daily calorie requirements of human population (Jan et al., 2001; Afrasiab and Jafar, 2011). Rice
breeders have mostly focused on utilizing natural diversity using hybridization and
recombination methods. Some of the rice cultivars including Taichung Native-I, IR-8, Mashuri,
IR-36, and other products of are well-known around the world for their better performance
against stresses. Despite this, breeders’ attention is occasionally drawn to technologies that
provide faster solutions to a wide variety of challenges (Raina, 1989). Among these
advancements, plant tissue culture techniques have received the most interest since they
promise to provide a variety of approaches to crop improvement difficulties.Tissue culture of
rice is one of the approaches of increasing rice production via efficient plant regeneration
method (Yaqoob et al., 2016; Kaya and Karakütük, 2018). Different explants are used to initiate
rice calli which include immature embryos, mature embryos, root segments, coleoptiles and leaf
bases that play a significant role in attempts to increase rice yield (Seraj et al., 1997; Li et al.,
2007; Benlioglu et al., 2015) (Figure 7).
Figure 7
Rice tissue culture (A–C) Single cell to callus formation (D, E) Sub culturing and regenerated
plants are ready to move to pots.
6.2 Pharmaceutical industry
Plants are used to treat and prevent particular illnesses and diseases in human beings since the
time immemorial (Chandran et al., 2020). Egyptians, Romans, and Chinese give strong proofs of
the usage of medicinal plants for the treatment different human ailments (Stafford,
1991; Cowan, 1999). Plants which possess healing metabolites with useful pharmacological
effects are referred to as medicinal plants (Atanasov et al., 2015). They are rich in
phytochemicals which have the spectacular capability to treat diseases and may be used for the
industrial production of pharmaceuticals and nutraceuticals (Chandran et al., 2020; Pant et al.,
2021). The medicinal properties of this flora are due to the heterogeneous group of herbal
metabolic merchandise called secondary metabolites which can be divergent of their structure
and metabolic pathways (Shasmita and Naik, 2018). The extensive research on plant cell culture
has caused a surge in the use of this technique in the pharmaceutical industry (Czyz et al.,
2014; Hayden et al., 2014; Mali and Chavan, 2016). The manufacturing of pharmaceuticals using
culture systems of plants can provide remarkable benefits including cost reduction, quick
production, and scalability (Hesami et al., 2017; Espinosa-Leal et al., 2018). Plants are abundant
sources of pharmaceutically significant compounds; however, there is a need to manufacture
these compounds within stringent laboratory conditions (Table 2).
Table 2
Plant-derived pharmaceuticals and their uses (Espinosa-Leal et al., 2018).
Plant-Made Plant Uses Manufacturer/notes
Pharmaceutical
ELELYSO™ Carrot or tobacco cell Enzyme Protalix, Carmiel, Israel and
(taliglucerasealfa) culture replacement Pfizer, USA, ProCellEx® Stable
Expression, First plant-made
human recombinant
therapeutic protein approved
(2014)
Vaccine (NDV) Tobacco suspension Against Dow Agrosciences, LLC,
cultures Newcastle Indianapolis, USA, First
disease virus tobacco cell-based vaccine
approved by the FDA against
Newcastle disease virus in
poultry
VEN150 Rice seeds For HIV- Ventria Bioscience, Junction
associated City, KS, USA, Express Tec
chronic Stable Expression Scale Cost
inflammation
Moss-GAA Moss Pompe disease Greenovation Biotech GmbH,
Heilbronn, Germany
Moss Physcomitrella patens-
based Broytechnology
Speed Scale and Customized
Moss-GBA Moss Gaucher’s Greenovation Biotech GmbH,
Moss-AGAL Nicotianabenthamiana disease; Fabry Heilbronn, Germany,
Alfalfa disease; Moss Physcomitrella
Vaccines Influenza, patens based Broytechnology,
Rabies Speed Scale and Customized
Rotavirus Medicago, Québec, QC,
Canada
Proficia™ Transient
Expression; Stable Expression
Antibody Duckweed leafy For non- Synthon, Nijmegen, The
biomass Hodgkin’s Netherlands, LEX system
lymphoma Stable expression, Speed
quality
Antibody Tobacco leaves HIV Fraunhofer IME, Aachen,
Germany, Stable Nuclear
Expression
Scale Cost
Serum albumin Rice seed Healthgen, Wuhan, Hubei,
Plant-Made Plant Uses Manufacturer/notes
Pharmaceutical
China, Stable Expression,
Quality Scale
CaroRx Tobacco leaves Dental caries PlanetBiotechnology,
Hayward, CA, USA, Stable
Expression
Quality Scale
PBI-220 Tobacco leaves Antibody for PlanetBiotechnology,
anthrax Hayward, CA, USA, Stable
DPP4-Fc Coronavirus Expression
infection Quality Scale
6.2.1 Secondary metabolite production from plant cultures
The tissue culture experiments on medicinal plants conducted by Veeresham and Chitti
(2013) revealed that various secondary metabolites having medicinal values can be obtained
from plant cell culture (Bhattacharyya et al., 2017; Shasmita and Naik, 2018). Biotechnological
approaches associated with plant tissue culture have increased the scope of medicinal plants
along with traditional agriculture used for the industrial production of bioactive metabolites
(Matsuura et al., 2018; Pant et al., 2021). Micro-propagation is a valuable technology since
many secondary plant metabolites cannot be manufactured chemically (Caldentey and Inze,
2004; Bhattacharyya and Van-Staden, 2016).
6.2.2 Plant cell suspension cultures
An extensive research is being carried out on exploring the biosynthetic properties of plant cell
cultures over the last decade (Siahsar et al, 2011). Cell suspension culture methods are
currently being employed for large-scale plant cell culture from which secondary metabolites
are extracted. A suspension culture is created by moving the comparatively friable component
of the callus into liquid media and maintaining it under appropriate physical conditions of
aeration, agitation, light, temperature, and other physical factors (Chattopadhyay et al.,
2002; Rao and Ravishankar, 2002). Cell cultures not only produce defined standard
phytochemicals in huge quantities, but they also reduce the presence of interfering substances
found in field-grown plants. The primary benefit of cell cultures is the production of bioactive
secondary metabolites in a controlled environment that is independent of climate and soil
conditions (Chattopadhyay et al., 2002).
6.2.3 Hairy root cultures
In the last two decades, this technique based on Agrobacterium rhizogenesinoculation has
gained popularity as a means of creating secondary metabolites generated in plant roots
(Palazon et al., 1998). Organized root cultures can contribute significantly to the generation of
secondary metabolites. Hairy root disease is caused by Agrobacterium rhizogenes in plants. The
neoplastic (malignant) roots, generated at rapid growth rate, by A. rhizogenes infection are
genetically stable and are developed in hormone-free conditions (Hu and Du, 2006). Hairy roots
high stability (Giri and Narasu, 2000) and productivity allow them to be used as a powerful
instrument for the recovery of important secondary metabolites (Pistelli et al., 2010). Generally,
substances released by roots serve as a stockpile of defense chemicals that plants can use to
strengthen their inherent defense mechanisms or defend themselves against pathogenic
attacks. In order to improve and optimize the number and quality of medicinal plant
metabolites, many plant tissue culture techniques have been extensively investigated (Pant,
2014). The following are some of the most often utilized plant-derived metabolites as
medicines.
6.2.4 Pharmacologically important plant secondary metabolites
2004; Bhattacharyya and Van Staden, 2016).
Tissue culture has been extensively utilized in breeding programs for over 50 years. The hybrids
of such crosses are often sterile due to embryo abortion but can be ‘rescued’ by means of
culturing or transplanting the embryos. The medium utilized for cell culture can be optimized
for the production of desirable products (Atanasov et al., 2015; Pant et al., 2021). Various plant
tissue culture systems have been extensively studied to improve and enhance the production
and quality of plant metabolites produced by the medicinal plants (Pant, 2014).
Alkaloids are the structurally diverse group of secondary metabolites which possess significant
biological activities (Moreira et al., 2018). Plants make them as a defense mechanism in
response to biotic and abiotic stressors (Taha et al., 2009), hence, several plants produce them
in reaction to caterpillars that feed on them, and they are potentially dangerous (Hussain et al.,
2018; Peng et al., 2019). The invasion species’ parasympathetic nervous system is blocked by
these alkaloids. Plant tissues would collect very little or no alkaloids if this sort of aggression did
not exist (Apone et al, 2010). The following are some of the most commonly utilized alkaloids in
the pharmaceutical business:
The isoquinoline alkaloid berberine (BBR) is extracted from the roots, stems, and rhizomes of
Coptis chinensis, Coptis japonica, and various other herbal plants and used in oriental
medication for hundreds of years ( Peng et al., 2019 ). Acupuncture, Chinese herbal medicines,
oriental nutrition, and dietary therapy, and tuina or oriental bodywork are the five major
branches of oriental medicine, which are the oldest codified system of medicine (Chen et al.,
2017; Yu et al., 2018). They restore health and balance by treating illness through five major
branches of oriental medicine, which include acupuncture, Chinese herbal medicines, oriental
nutrition and dietary therapy, and tuina or oriental bodywork. Plant extracts are used to make
Oriental remedies (berberine chloride) (Zhang et al., 2017). Moreover, berberine is typically
used as an oral medicine for the treatment of excessive cholesterol, high blood pressure,
diabetes, or excessive levels of lipids in the blood (Kawano et al., 2015; Jing et al., 2018). It is
also used as a topical remedy for canker sores and burns (Cui et al., 2018).
Valepotriates is the name given to a set of compounds determined to have tranquilizing results,
in addition to proof of having antitumor and cytotoxicity effects (Boss et al., 2002; Patocka and
Jakl, 2010). Valerianaceae, consisting of herbs and rarely shrubs, has been used for decades to
obtain medicinal drugs. Traditionally, valepotriates has been used to treat spastic colitis and
gastrointestinal pain (Becker and Chavadej, 1988; Andreatini and Leite, 1994). The tissues
of Valerianaceae,placed in a growth medium, are used to investigate which species produced
better levels of the desired compounds and are resistant against various stresses. A lot more
research is being done on the micro-propagation of Valerianaceae to explore the compounds
with medicinal value (Boss et al., 2002; Jarema, 2008) (Figure 8).
Figure 8
Structures of (A) Berberine (B) Valepotriates (C) Taxol.
Another compound is Taxol which was generically known as paclitaxel, was granted FDA (Food
and Drug administration) approval in 1992 (Khosroshahi et al., 2006). The drug is administered
to ovarian cancer patients and is often used as a secondary treatment when chemotherapy is
failed in these patients (Badi et al., 2015). Taxol is mainly extracted from the bark of wild Taxus
brevifolia trees, however, due to its increased demanda as an anti-cancerous drug; researchers
are primarily interested in its in vitro propagation of taxol producing flora for a more rapid and
environment friendly technique (Walker et al., 2004; Nasr et al., 2007). As a result, plant tissue
culture is a viable technology for producing desirable bioactive chemicals from plants. Plant
tissue culture is also used to help save endangered species, as many therapeutic plants are on
the verge of extinction due to overuse (Rout et al., 2000; Bestoso et al., 2006; Chen et al.,
2016c; Vidyagina et al., 2021). Further investigations on various plant species having medicinal
values is needed to explore as most of the natural flora is still unidentified. The development of
plant tissue culture techniques will expand the long-term use of therapeutic plants in the future
(Balunas and Kinghorn, 2005).
6.3 Cosmetic Industry
Over the last ten years, the enticing trend of natural cosmetics production has ushered in a new
era of plant cell culture technology and during this time, more than 50 cosmetic products based
on extracts of plant cell cultures have been developed, the bulk of which are made with plant
cell suspension cultures (Charles et al., 2017; Krasteva et al., 2020). Plant cell culture cosmetic
production is not dependent on appropriate seasonal conditions; thus, it requires less time and
energy. Cosmetic extracts derived from plant cell cultures suit the market’s increasingly
stringent safety requirements. In addition to being free of pathogens, pollutants, and pesticide
residues, plant cells generated under aseptic laboratory conditions rarely include any malignant
compound or potential allergen, which would otherwise destroy the majority of the plant
extracts obtained (Schmid et al., 2008; Trehan et al., 2017; Eibl et al., 2018). Plant cell
suspension cultures are cultivated in single-use wave-mixed bioreactors or renewable stainless
steel stirred bioreactors in commercial production today. Single-use wave-mixed bioreactors are
perfect for personalized goods due to their modest operational capacity, improved security, and
more quick and simple operation (Barbulova et al., 2014; Krol et al., 2020).
In recent decades, due to increased customer demand for cutting-edge cosmetic formulations
made with efficient, secure, and sustainable components, the cosmetic industry has expanded
internationally and become extremely competitive (Zappelli et al., 2016). Plant tissue cultures
are a perfect source of safe and pure components for cosmetic goods since they can be
cultivated under controlled conditions with minimum possibility of pathogen or environmental
contamination. Utilizing various extraction techniques and solvents while taking advantage of
the chemical makeup of plant cell components, plant tissue culture technology allows the
isolation of many active ingredients from a single culture (Apone et al., 2018). Discussed below
are in vitro propagated plant species specifically known for producing valuable
pyhtocompounds with potential uses in cosmetics: (Table 3).
Table 3
Popular active cosmetic ingredients derived through plant cell culture technology (Krasteva
et al., 2020).
Product Plant Active Applicatio Manufac Web page
Source compou n(s) turer
nd(s)
PhytoCell Symphyt Water Boosts the Mibelle https://
Tec™ um extract regenerativ AG mibellebiochemistry.com/
Symphytu officinal e power of Biochemi phytocelltectm-symphytum
m e L. epidermal stry
stem cells,
increases
epidermal
thickness,
improves
barrier
function
PhytoCell Saponari Water Maintains Mibelle https://
Tec™ a extract dermal AG mibellebiochemistry.com/
nunatak® pumila L. stem cell Biochemi phytocelltectm-nunatakr
vitality stry
after UV
irradiation,
improves
skin
density,
Product Plant Active Applicatio Manufac Web page
Source compou n(s) turer
nd(s)
elasticity
and
firmness
Deobiom Morinda Anti‐ Protection Vytrus https://www.vytrus.com/
e Noni citrifolia quorum and Biotech products/deobiome-noniprcf-the-
L. sensing balance of biological-deodorant/?
molecule microbiom portfolioCats=17
s; sugars e,
Metabolis
m shift
from lipids
to
polysaccha
rides,
Quorum-
sensing
inhibitors,
Skin health
and
detoxificati
on
Arabian Gossypiu Phenols Anti‐ Vytrus https://www.vytrus.com/
Cotton m and oxidants, Biotech products/arabian-cottonprcf-the-
PRCF herbace flavonoid Photoprote broad-spectrum-protector-
um L. s ctive (sun- against-photo-aging/?
block), portfolioCats=17
enhances
cell
viability,
modulates
inflammat
ory
response
in human
epidermal
progenitor
cells
6.3.1 Bean (Dolichos biflorus)
A hydrosoluble extract of Dolichos biflorus cell cultures was examined for the amount of
isoflavones such genestin and daidzen, and their glucosidic derivatives. In addition to avoiding
cellular damage, these substances also reduce the inflammation caused by ultraviolet (UV) light
in dermal and epidermal cells, known as solar erythema (sunburn), which is characterized by
skin reddening brought on by an increase in blood flow and capillary dilation (Bimonte et al.,
2014).
6.3.2 Shrub (Daphne odora)
Another illustration of the characterization of a plant cell culture extract for cosmetic uses
comes from the evergreen shrub Daphne odora, which is particularly interesting because it can
withstand low temperatures. The ability of a hydrosoluble extract from shrub cell cultures to
alleviate cutaneous irritation by cold stress was investigated (Bimonte et al., 2018).
6.3.3 Raspberry (Rubus ideaus)
The expression of genes related to skin hydration, including aquaporin 3, filaggrin, and
involucrin, was induced by the liposoluble extract made from raspberry cell cultures.
Additionally, the extract boosted the expression of the ceramide-producing enzymes
glucocerebrosidase and hyaluronic acid synthase and stimulated their activity. Additionally, it
demonstrated an extraordinary capacity to hydrate skin when tested on human skin in vivo,
indicating that it had tremendous potential as a skin care agent, particularly for dry and ageing
skin (Titos et al., 2015).
6.3.4 Apple (Malus domesticus)
The hydrosoluble and liposoluble fractions of Malus domesticus (apple) cell suspension cultures
were combined, the entire cell lysate was subjected to high-pressure homogenization, and they
were then added to the finely dispersed liposomes to create a cosmetic ingredient
(nanosomes). The expression of the antioxidant enzyme Heme Oxigenase 1 was first examined
to gauge the preparation’s anti-senescence effects. Four weeks of treatment resulted in a 16%
reduction in wrinkle depth, according to clinical studies (Schmid et al., 2008). Additionally, it has
been demonstrated that this extract is excellent at shielding human stem cells from UV rays.
6.3.5 Rose mallow (Hisbiscus syriacus)
Another example of an active ingredient created for use in skin care products comes from rose
cell suspension cultures (Di Martino et al., 2017). Flavonoids and coumarins, which are known
for having tissue-regenerating capabilities, were found in the hydroethanolic extract of H.
syriacus cellsin a chemical study (Borges Bubols et al., 2013). On both fibroblasts and
keratinocytes, the extract was examined for its capacity to hydrate wounds and promote wound
healing. Ex vivo studies of human skin biopsies from injured people confirmed these findings in
which topical applications of H. syriacus extract dramatically improved wound closure by
increasing the production of neo-epidermis (Di Martino et al., 2017)
6.3.6 Butterfly bush (Buddleja davidii)
In plant cell suspension cultures of the butterfly bush (Buddleja davidii), the chemical
verbascoside, a phenylpropanoid glycoside, renowned for its antioxidant, anti-inflammatory,
photoprotective, and chelating activities was generated in high concentrations (Vertuani et al.,
2011). In vivo investigations showed that this substance suppressed the activity of collagenases
linked to skin ageing as well as the activation of pro-inflammatory factors.
6.3.7 Coffee (Coffea bengalensis)
Bengal coffee (Coffea bengalensis) plant cell culture extract was created as an active ingredient
to support skin health and homeostasis because it did not include the alkaloid caffeine.
Caffeine’s anti-obesity properties increase its utility in cosmetics. However,it has been
investigated that oral use of caffeinedecreased the thickness of hypodermis, which is linked to
wrinkle formation.
6.3.8 Trefoil (Lotus japonicas)
In a recent study, a peptide/sugar mixture that was extracted from the cell walls of cultures
enriched with somatic embryos of Lotus japonicus plant was considered as a possible skin-
rejuvenating element (Tito et al., 2019). When the combination was chemically characterized, it
was discovered to include a significant sugar fraction with high concentrations of glucose,
galactose, mannose, and fructose. Saccharides are recognized to have advantageous effects on
hydration with anti-inflammatory effect on dermal cells. As a matter of fact, the function of
sugars in cosmetics has not been fully understood, more research needs to be done to explore
the underlying mechanism (Tolg et al., 2014 ).
Plant cell cultures are now being used for the production of ‘cosmeceuticals’, products having
cosmetic as well as therapeutic (medical or drug-like) effects that exert beneficial effects on skin
health. An extensive investigation is being done to investigate the plant sources producing
active ingredients, such as antioxidants, ingredients with antimicrobial, anti-viral, anti-
cancerous, anti-fungal, anti‐inflammatory, and anti-allergy properties along with moisturizing,
anti‐ageing, anti‐wrinkle and UV protective properties, which are crucial to cosmetics industry
(Apone et al., 2010; Tito et al., 2011; Morus et al., 2014). Most of the phytochemicals such as
polyphenols, phenolics acids, triterpenes, flavonoids, stilbenes, steroids, carotenoids, steroidal
saponins, sterols, fatty acids, polysaccharides, sugars, and peptides are extracted with relevant
solvents and utilized as active constituents in cosmetic preparations (Barbulova et al.,
2014; Furusaki and Takeda, 2017).
Applications of plants/flowers extracts in cosmetics are significant which include skin
moisturizing, whitening or tanning products, sunscreens, radical-scavenging antioxidants,
immune stimulants, and skin thickeners etc. (Ochoa‐Villarreal et al., 2016; Georgiev et al.,
2018). Because of its ability to minimize wrinkles in the crow’s feet area of the face, a liposome-
encapsulated extract of cultured apple stem cells is employed as an ideal ingredient in anti-
aging products (Satish et al., 2019; Krasteva et al., 2020; Krol et al., 2020). Chemicals isolated
from Catharanthusroseus are currently used in the manufacturing of both ordinary consumer
and professional care cosmetics, demonstrating significant potential in protecting skin from
heavy metal toxicity (Mallurwar and Pathak, 2008; Schmid and Zülli, 2012; Blum et al.,
2013; Hayta et al., 2017; Meir et al., 2018). Moreover, the supplements that lessen hair loss and
skin aging by nourishing skin and boosting body’s immune system are being produced via plant
tissue culture (Ribeiro et al., 2015; Murthy et al., 2019). The concentrates from unique or
endangered plant species can also be made accessible for cosmetic production using plant cell
culture techniques (Blum et al., 2013; Ribeiro et al., 2015; Chen et al., 2016b; Murthy et al.,
2019). Melanin is one of the most extensively dispersed pigments found in bacteria, fungi, and
animals, therefore, suppression of the melanin-producing enzyme tyrosinase is one of the most
frequent defense strategies used by the roots (Apone et al, 2018). Human skin color is largely
influenced by the amount of melanin produced by melanocytes, and tyrosinase controls this
pigment manufacture. Thus, one of the primary objectives of skin lightening substances in
cosmetics is to reduce the effects of sun exposure and age spots, which are mostly constituted
of melanin and give the skin an even tone. Sena et al. (2018) examined the depigmentation
effects of two different extracts made from the hairy root cultures of, subsp. pekinensis (Chinese
cabbage) and Brassica Rapa. Some popular plant-derived active cosmetic ingredients that are
currently available in the market as mentioned and elaborated in Table 3.
Go to:
7 Limitations
Despite of various useful applications of plant-derived compounds obtained via in vitro plant
propagation, there are certain limitations which need to be addressed in order to reap
maximum benefit of this technique. The limitations include difficulties with continuous
operation, product removal, and aseptic conditions. A few culture systems appear to have the
potential to become commercially viable because of these limitations (Goel et al., 2011).
Furthermore, the prevalence of somaclonal variation in populations formed from tissue culture
has a detrimental impact on the utilization of tissue culture and has remained a serious
concern. Somaclonal variation is variance that originates in cell and tissue cultures. At the
moment, the word somaclonal variation refers to all types of tissue culture produced variations
(Bajaj, 1990), Since Braun (1959) first observation and description of somaclonal variation; it has
been one of the key issues of many tissue cultivated plants. Plant cell development in vitro and
regeneration into full plants is an asexual process that involves just mitotic division of the cell
and, ideally, should not result in variation. Clonal multiplication of genetically homogenous
plants is the ideal scenario. Uncontrolled and unpredictable spontaneous variation throughout
the cultural process is thus an unanticipated and largely undesirable phenomenan (Ranghoo-
Sanmukhiya, 2021). This is attributed to somaclonal variation in production clones and low
secondary metabolite titers (Sharma et al., 2014). In contrast to these detrimental
consequences, its use in crop improvement through the development of new variations is
widely recognized.
The expense of culture material, electricity, and labor are other issues with in vitro tissue
cultivation. Alternative materials such as home sugar or other sugars as carbon sources, as well
as various types of starches and plant gums in place of agar, have been used in numerous
experiments to overcome this problem. Alternatives have included liquid media and cell
suspension cultures, temporal immersion systems, and reusable glass beads as support matrices
(Etienne and Berthouly, 2002; Ahloowalia and Savangikar, 2003; Sahu and Sahu, 2013; George
and Manuel, 2013). Bioreactors and robotic propagule handling, for example, have been shown
to save production costs. Another issue that occurs with plant tissue culture is the plants’
genetic stability. Somaclonal differences that occur during in vitro propagation, commercial
phytochemical synthesis, or genetically modified plants can have significant economic
consequences and are a major impediment to the practical application of plant tissue culture
techniques for the production of active metabolites (Rahman and Rajora, 2001). As a result, the
genetic constitution and stability of in vitro-regenerated plants must be monitored and
examined in order to screen somaclonal variability within a cell culture. As part of the
techniques utilized in that process, several strategies are used to analyze possible adjustments
at various levels (Bhattacharyya and Van-Staden, 2016).
Go to:
8 Conclusion and future perspectives
Plant micro propagation is a powerful technique in order to acquire plant extracts with various
commercial applications than using whole plants. The application of targeted genome
engineering, notably the previously stated genome editing mediated by CRISPR/Cas9, is one of
the most important techniques. Using this technical approach, new plant kinds can be created
without the input of foreign genes (Doudna and Charpentier 2014; Baltes and Voytas
2015; Nogueira et al., 2018). Furthermore, the cosmetics sector which introduces hundreds of
new cosmetics items every year is heavily influenced by customer demand. Plant cell culture
extracts with several particular actions for skin care, make-up, and hair care as supplement
components are gaining popularity in the cosmetics sector. Consumers desired cosmetics that
are effective, safe and natural can be obtained by exploring phytochemicals with these desired
properties under in vitro conditions. The development of efficient and appealing novel active
ingredients for cosmetics, and skin care in particular, is developing in this direction. In this
regard, plant tissue culture methods hold enormous promise. In the future, plant tissue cultures
might not only be a source of novel chemicals with uncharted biological actions, but they might
also work as alternative recombinant protein biofactories, particularly for those whose
expression might be problematic or constrained in fermenting microbes. In the next decade,
tissue culture technique should attain its full potential, thanks to new technologies like gene
editing and environmental component manipulation. However, the major constraints need to be
addressed for sustainable industrial applications of in-vitro regenerated plants on a large scale.
2004; Bhattacharyya and Van Staden, 2016).
Go to:
Author contributions
AH, SA, FK, ME, SN, AA and AB designed and wrote the manuscript. MH, GM and SI have done
graphical work. AB, RA, DM and MM helped to revise the manuscript. AH, SN, SI, AB, MH and
MA critically revised and supervised the manuscript. RA and DM provided the funds for the
publication of this review article. All authors contributed to the article and approved the
submitted version.
Go to:
Acknowledgments
We thank the Faculty of Process and Environmental Engineering Łódź University of Technology,
Poland for providing financial support to the current study.
Go to:
Conflict of interest
The authors declare that the research was conducted in the absence of any commercial or
financial relationships that could be construed as a potential conflict of interest.
Go to:
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