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156 CHAPTER 9 Rhodiola imbricata
https://t.me/med1917
Table 9.4 Amino acid content in root extract of Rhodiola imbricata (Tayade
et al., 2017).
S. no. Amino acid Abbreviation Content (mg/g)
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
13.
14.
15.
16.
17.
18.
19.
20
21
BDL, below detection limit; ND, not detectable.
L-2-amino-n-butyric acid Abu ND
L-Alanine Ala 1142.33 11.02
L-Arginine Arg Arg 214.67 7.09
L-Aspartic acid Asp 434.67 8.74
L-Cystine Cys 239.33 8.39
L-Cystine HCl Cys HCl 1136.33 11.72
L-Glutamic acid Glu 320.67 7.77
L-Glycine Gly 1640.67 11.85
L-Histidine His 1434.33 10.02
L-Isoleucine Ile 91.33 7.77
L-Leucine Leu 928.67 10.79
L-Lysine Lys 1329.33 11.55
L-Methionine Met 736.67 8.02
L-Nor Leucine Nor Leu 1038.67 10.21
L-Ornithine Orn ND
L-Phenylalanine Phe 855.33 9.02
L-Proline Pro 1263.67 10.50
L-Serine Ser 839.67 10.97
L-Threonine Thr 1015.67 8.02
L-Tryptophan Trp ND
L-Valine Val BDL
Table 9.5 Mineral content of Rhodiola imbricata root (Tayade et al., 2017).
S. no. Minerals Symbols Content (mg/kg)
1. Calcium Ca 11034.17 332.04
2. Chromium Cr 7.27 0.32
3. Cobalt Co 2.98 0.06
4. Copper Cu 3.49 0.12
5. Iron Fe 1441.17 27.98
6. Magnesium Mg 581.99 17.40
7. Manganese Mn 75.78 2.21
8. Molybdenum Mo 2.65 0.05
9. Nickel Ni 4.89 0.21
10. Phosphorous P 376.72 11.87
11. Potassium K 2143.25 65.37
12. Sodium Na 109.75 3.32
13. Zinc Zn 16.27 0.54
9.4 Pharmacological properties 157
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9.4.1 Adaptogenic and antifatigue activity
Adaptogens are substances that enable the standardization of physiologic responses
to various stresses, increase the stress tolerance of the body, and enhance work performance (Darbinyan et al., 2000; Grace et al., 2009). Rhodiola extracts have great
utility in treating esthetic conditions which develop after intense physical or intellectual strain, including a decline in work performance, sleep difficulties, irritability,
poor appetite, high blood pressure , headaches, and fatigue. Thus, it is effective in
preventing oxidative stress following exhaustive exercise.
9.4.2 Elongation of life span and antiaging activity
An animal study found that Rhodiola extract could inhibi t the death of thymic
T cells, which is important as thymus function decreases with age. It reversed
D-galactoseeinduced aging effects in neural and immune systems, improved motor
activity, increased memory latency time, and enhanced lymphocyte mitogenesis and
interleukin-2 production (Jafari et al., 2007; Mao et al., 2010a,b).
9.4.3 Antioxidant properties
Singlet oxygen scavenging, H2O2scavenging, hypochlorite scavenging, ferric
reducing, ferrous chelating, and protein thiol protection activities were noted
(Chen et al., 2008). Salidroside reduced hydrogen peroxideeinduced intracellular
Reactive Oxygen Species production in human erythrocytes. Salidroside also
increased cell survival and prevented human erythrocytes from undergoing eryptosis
or erythroptosis mediated by H
(Qian et al., 2012).
2O2
9.4.4 Antidepressant
It was noticed that orally administering salidroside for 2 weeks increased olfactory
bulbectomyeinduced hyperactivity in an open-field test and reduced immobility
time in a forced swimming test. Reduction in TNF-a and IL-1b levels in the hippocampus was also noted. Salidroside also increased glucocorticoid receptor and
brain-derived neurotrophic factor expression in the hippocampus of rats. In addi tion,
salidroside attenuated corticotrophin-releasing hormone expression in the hypothalamus and the levels of serum corticosterone (Yang et al., 2014).
9.4.5 Skin treatments
Salidroside inhibited UVB-induced hyperpigmentation in brown guinea pig skin by
reducing the number of DOPA-positive melanocytes in the basal layer of the
epidermis and reducing tyrosinase activity and melanin synthesis in melanocytes
(Peng et al., 2013). R. rosea extract, salidroside, and tyrosol may be effective
skin-whitening agents.
158 CHAPTER 9 Rhodiola imbricata
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9.4.6 Analysis of heavy metal
Studies have shown that the concentration of heavy metal in aqueous extracts of
R. imbricata, except chromium, was less than the maximum permissi ble ranges proposed by the World Health Organization.
9.4.7 Radioprotective efficacy
In a study conducted on protection, lethal gamma irradiation (10 Gy) induced mortality in Swiss albino strain “A” mice. Preirradiation administration of extracts produced 83% to more than 90% survival, beyond the 30 days of observation period.
The studies suggested that R. imbricata is a suitable radioprotector of herbal origin
(Goel et al., 2006).
Hydroalcoholic rhizome extract of the plant showed antihemolytic capacity by
preventing radiation-induced membrane degeneration of human erythrocytes. The
study showed that R. imbricata rhizome renders in vitro and in vivo radioprotection
via multifarious mechanisms (Arora et al., 2005).
9.4.8 Adjuvant activity
Antiinflammatory or immunosuppressive effect of R. imbricata rhizome was tested
in adjuvant-induced arthritis model. The study suggested that the plant rhizome has
adjuvant/immunopotentiating activity in terms of cell-mediated as well as humoral
immune response (Mishra et al., 2010).
9.4.9 Mechanism and action of poststress caused by hypothermia
induced by cold, hypoxia, and restraint (C-H-R) stress
Doses of extract administered 30 min prior to induced C-H-R stress and hypothermia induction decreased or maintained tissue glycogen and enzyme activities,
viz., PFK, CS, G6-PD, and HK, in liver blood and liver, on attaining T(rec) 23
and recovery. The resu lts suggest that R. imbricata extract treatment in rats shifted
anaerobic metabolism to aerobic, during C-H-R exposure and poststress recovery
(Gupta et al., 2009).
Other physiological effects of Rhodiola and Salidroside include the following
(Chiang et al., 2015):
• Antiinflammatory
• Protection against neuron damage
• Liver protection
• Reduction of oxidative stress in cardiovascular diseases
• Used in diabetes mellitus
• Obesity
• Antiviral
• Used against lung cancer
C
9.5 Cultivation and propagation of Rhodiola imbricata 159
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Salidroside protects human erythrocytes by its antioxidant activity and caspase-3
inhibition in a dose-dependent fashion (Qian et al., 2012). It also protects hematopoietic stem cells from oxidative stress by activating PARP1, a DNA repair enzyme
actively involved in cell apoptosis (Li et al., 2014). Results of various clinical trials
have revealed that Salidroside possesses various functions such as anticold, antifatigue, antianoxic, antivirus, antimicrowave radiation, and antit umor. It also possesses various medicinal properties such as preventing illness associated with old
age, strengthening attention spans, delaying senility, and improving work efficiency.
Due to its environmental acclimation activity, it plays important roles in healthcare,
military, sports, and aerospace.
In the phytochemical study, the content of salidroside, rosavin, and its derivatives
in Rhodiola plants depends on the morphological parts of the plant as if extracted
from roots or rhizomes. In addition, it also depends on the age and sex of the plant,
where the male rhizomes of the plants accumulated higher amounts of salidroside in
R. rosea than their female counterparts (Platikanov and Evstatieva, 2008; Weglarz
et al., 2008). The location and timing of collection also influence the salidroside con-
tent of the plants (Bykov et al., 1999). To date, document data related to the chemical
profile of Rhodiola plants have shown that samples taken from natural sources have
a higher salidroside and rosin content than in the cultivated fields.
9.5 Cultivation and propagation of Rhodiola imbricata
As the highest medical importance has been associated with secondary metabolites
present in the roots of the plant, according to the culture experiments performed in
southern Finland, Rhodio la spp. can be efficiently grown using organic plant culture
methods (Tayade, 2015). For the natural stratification of winter, the seeds should be
planted in the fall to produce seedlings. For approximately 1 year before transplant,
these seedlings should be kept in pots due to the slow growth of the plants during the
first 2e3 years. After 4 years of planting, the first root yield is harvested. The weight
of the root and the yield of the roots of the plants depend to a great extent on the age
(Fig. 9.1).
R. imbricata plant is propagated by two methods, namely sowing seeds and
the cutting of rootstocks. In the general practice performed in tissue culture labs ,
60%e65% of the germination of the seed is done in the fields, while in the case
of the plantation of rootstocks, the scientists achieve a very high percentage of
survival ranging between 85% and 90%. Therefore, better rates of dispersion have
been achieved through the division of rootstocks. The plants of 3e5 years are
considered ideal for the plantation of appropriate rootstocks (Sharma, 2016).
For micropropagation of R. imbricata, generally the proportionate concentration
and combinations of Auxin, Cytokinin, and Gibberellic acid (like Indole-3-butyric
acid, Kinetin, 6-Benzylaminopurine, GA
cessful of these being MS media supplemented with BAP (1 mg/L) þ IBA (2 mg/L)
for multiple shoot culture (Pundir et al., 2019 )(Figs. 9.2e9.4), while BAP (2 mg/
L) þ IBA (4 mg/L) was found good for root and shoot induction and growth
(Sharma, 2016).
, and Thidiazuron) are used. The most suc-
3
160 CHAPTER 9 Rhodiola imbricata
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FIGURE 9.1
Rhodiola imbricata from Trans-Himalayan cold desert of Ladakh region, India (Tayade,
2015).
FIGURE 9.2
Micropropagated plants of Rhodiola imbricata growing in MS media supplemented with
BAP (1 mg/L) þ IBA (2 mg/L) (Pundir et al., 2019).
9.5 Cultivation and propagation of Rhodiola imbricata 161
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FIGURE 9.3
Roots induced in micropropagated plant growing in MS media supplemented with BAP
(2 mg/L) þ IBA (4 mg/L) (Sharma, 2016).
FIGURE 9.4
Callus of Rhodiola imbricata liquid MS media with BAP (1 mg/L) þ IBA (2 mg/L) (Pundir
et al., 2019).
162 CHAPTER 9 Rhodiola imbricata
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Tasheva and Kosturkova (2010), developed efficient schemes for the regenera-
tion and micropropagation of R. rosea. Zhao et al. (2012), developed a modified temporary immersion bioreactor with forced ventilation that reduces the rate of
hyperhydration and improves the quality of outbreaks and multiplication rates in
R. crenulata. The similar approach was used for R. imbricata where plant was
micropropagated in liquid suspension culture containing MS media supplemented
with BAP (1 mg/L) þ IBA (2 mg/L) (Pundir et al., 2019). Th e idea behind using
liquid suspension lies in short growth time and ease in multiplication of plant.
Zych et al. (2005), showed a successful encapsulation of differentiated callus and
axillary buds in calcium alginate beads in Rhodiola kirilowii. These capsules were
able to grow in shoots and seedlings in solid basal MS medium. This is yet to be
explored in R. imbricata species.
Another significantly important approach used to seize the medical viability of
plant in an efficient and immediate basis is using its callus culture for either directly
attaining potential secondary metabolite or using callus as substitute of explants for
micropropagation. For this purpose, MS media supplemented with TDZ (1 mg/L) is
found most appropriate (Pundir et al., 2019) Fig. 9.5.
In recent years, a number of approaches have been used in order to increase content of pharmaceutically important secondary metabolites. Kapoor et al. (2018) ,
studied the potential effect of light quality on biomass accumulation and production
of industrially important secondary metabolites in callus cultures of R. imbricata.
The results indicated blue light as promising light source for the enhanced production of flavonoids, secondary metabolite salidroside, and phenolics content in callus
cultures.
FIGURE 9.5
Callus of plant growing in MS media supplemented with TDZ (1 mg/L) (Pundir et al.,
2019).
9.6 Genetic diversity 163
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In order to enhance the secondary metabolite production in R. imbricata cultures,
various elicitors were tested. Jasmonic acid enhanced salidroside production, ascorbic acid content, total flavonoid content, and total phenolic in Callus Aggregate Suspension cultures. DPPH-scavenging activity and total antioxidant capacity were also
enhanced upon Jasmonic acid treatment (Kapoor et al., 2019). Besides, treatment of
shoot cultures for ultraviolet (UV) light for 30 min was found as a promising physical elicitor for providing growth along with optimized medium in R. imbricata
shoot cultures (Pundir et al., 2019) Figs. 9.2 and 9.3.
9.6 Genetic diversity
In a study conducted by Gupta (2012), techniques like RAPD and ISSR markers
were used to characterize and compare the genetic diversity in three collected populations of R. imbricata. The genetic closeness among the Khardung La and Chang
La plants can be interpreted by the high rate of commonness in their individuality.
The genetic similarity among these individuals is probably linked with their resemblance in their genomic and amplified region. AMOVA used for RAPD, ISSR, and
RAPD þ ISSR-combined markers were used for testing genetic variation. Results
tabulated below showed significant (P < .001) genetic variation within population
than among populations.
S.
no. Markers
1. RAPD 56 44
2. ISSR 78 22
3. RAPD þ ISSR 71 29
Variation within
population (%)
Variation among population
(%)
Similar reports have been made in ISSR studies of populations of R. crenulata
(Lei et al., 2006), Rhodiola chrysanthemifolia, and R. alsia (Xia et al., 2005,
2007) which may be because of isolation of populations. Yan et al. (1999), reported
Rhodiola sachalinensis had high genetic diversity within population at high altitude
than that of growing at lower altitude.
As the plant grows in the Trans-Himalayan region of Ladakh, which is situated at
more than 3000 m above mean sea level, there are numerous factors which can lead
to partitioning of total genetic variation of a plant species which is different from the
general pattern. Factors such as the low temperature, high UV radiations, the insufficient content of oxygen, short vegetation period (approximately 120 days) make
the chance of seedling recruitment difficult and rare. Seed dispersal pattern and
germination of Rhodiola at Ladakh and other Trans-Himalayan regions have irregular dispersal pattern which can be related to the effect of wind and other stressful
habitat conditions (Gupta, 2012).
164 CHAPTER 9 Rhodiola imbricata
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9.7 Omics
Secondary metabolites are often associated with having enormous curative properties. In R. imbricata, Salidrosideda Phenylethanol derivative and Rosavind
Phenylpropanoiddhas been extensively studied for its various medicinal properties.
The biosynthesis of salideroside and rosavin involves numerous important genes
which have been illustrated below. The scientific community is making considerable
conscious efforts in either stimulating or enhancing the functionality of these genes
through number of bimol ecular studies which have been highlighted in Figs. 9.6 and
9.7 defining the biosynthetic pathway of the metabolites.
FIGURE 9.6
Propounded pathway responsible for biosynthesis of Salidroside (Ma et al., 2008).
9.7 Omics 165
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FIGURE 9.7
Pathway responsible for the biosynthesis of Rosin, Rosavin, and Rosarin (Grech-Baran
et al., 2015).
9.7.1 Salidroside
• Tyramine precursor of tyrosol governed by tyrosol
glucosyltransferase (TGase)
• Increased biomass and salidroside accumulation
were achieved by medium supplementation with
tyrosol than that in Rhodiola sachalinensis
• Increased salidroside by overexpression of the
UGT73B gene in R. sachalinensis
• Uridine diphosphate (UDP)-glucosyltransferase
cDNA (UGT72B14) expression resulted in high
salidroside production in vitro and in vivo in
R. sachalinensis
• TyrDC gene isolated from Rhodiola rosea had
expression with the accumulation of salidroside
• RcTyrDC enhanced the tyramine, tyrosol, and
salidroside biosynthesis
Xu et al. (1998), Yu et al.
(2011), Zhang et al. (2011)
Zhou et al. (2007)
Ma et al. (2007)
Yu et al. (2011)
Gyo¨rgy et al. (2009)
Lan et al. (2013)
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