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antagonistic cross-talk amongst the major stress-respons ive phytohormones which
include SA, JA, ABA, and ET along with other signaling phytohormones like BR,
CKs, AUXs, GAs, strigolactones, and nitric oxide play a vital role in plants to adapt
and develop tolerance to the ambient freezing temperature (Shi et al.
2015; Peleg and
Blumwald 2011).
11 Signaling Molecules in Medicinal Plants Response to Cold Stress 181
Abscisic
Acid
ABA,
isoprenoid
horm
o
ne
produced during plastidal 2-C methyl
D-erythritol-4-
phosphate pathway hold a prominent role in seed dormancy, stomatal regulation,
biogenesis of lipids and storage proteins, regulation of ambient/environmental
stresses, hydration status, and others. Due to low temperature the levels of ABA
significantly shoot up due to the activation and overexpression of ABA-regulated
genes via two cis-acting elements known as ABA-responsive elements (ABREs),
activated by AREB/ABF transcription factors and CRT/D RE, activated by MYB96–
HHP complexes, ICE1, and CAMTA (Yoshida et al. 2010; Eremina et al. 2016). As
a part of the defens e mechanism, ABA accentuates the genesis of ROS consequen-
tially leading to the accumulation of several antioxidant and non-antioxidant defense
mechanisms leading facilitating cold acclimation (Xue-Xuan et al.
2010). Panax
ginseng, (aka. Asian or Korean ginseng) a traditional herb utilized in Korea and
China for the medicament of inflammatory diseases, CVDs, Diabetes, Cancer, etc.
The active secondary metabolite that elates its therapeutic property is called
ginsenoside whose biosynthesis is greatly influenced by the biotic and abiotic
stresses (Kim
2018). T. hemsleyanum (Aka. Natural plant antibiotic), a traditional
Chinese herbal medicine taken in the form tea or dietary supplement play a predom-
inant role in the treatment of fever, convulsion, pneumonia, asthma, rheumatism,
hepatitis, PCOD, scrofula, etc. Alkaloids, polysaccharides, and flavonoids released
in both tubers and areal parts of the plant are of substantial interest (Peng et al.
2021).
High-altitude Himalayan Alpine Herbs Bistorta affinis (Aka. Border jewel) and
Sibbaldia procumbens are perennial plants investigated widely for their curative
effects for cold, cough, GIT disorders, and other conditions (Rahman et al. 2019).
The pharmaceutically active medicaments in these plants are greatly infl uenced by
cold stress and cold-stress induced desiccation. In this regard, ABA act as a distant
signalling molecule and elicits antioxidant systems preventing oxidative damage,
modifies stomatal oscillations and reduces transpiration and regulates the osmotic
homeostasis.
Gibberellin
Oxidative enzymes like GA 20-oxidases, KAO-oxidases, and GA 3-oxidases from
trans-geranylgeranyl diphosphate produce bioactive GAs in plants and they are
catabolized via GA 2-oxidases and feedback inhibition of the biogenetic pathway.
The catabolic pathways of GA are further governed by negative signaling regulators,
DELLA belonging to the GRAS protein family that act immediately after the
interaction of GA to its soluble receptor (GID1) and bring about its degradation
via Ubiquitin proteasomal pathway (Schwechheimer
2012). Also, they are
repressors of PIF3, PIF4, BRI1, and BES1/BZR1 transcription factors, play a key
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role in the anabolic pathways of bioactive GA (Feng et al. 2008; Li et al. 2012). This
bioactive hormone plays a critical role in the regulation of membrane fluidity,
osmolyte homeostasis, activation/over-expression of genes encoding vital enzyme
systems, etc., and thereby facilitates plant defense against cold stress (Emamverdian
et al.
2020). Species belonging to the genus Nicotiana are well known for their
catastrophic effects on health which has led to the negligence of traditional pharma-
copeia. Back in the day, the concoctions of the green leaves and roots of Tobacco
were widely employed in traditional Ayurvedic therapies for the treatment of
ulcerated abscesses, fistulas, sores, polyps, purgative, and narcotics (Charlton
2004). Lycopene, a non-provitamin A carotenoid is one the active SMs released
by tomatoes manifesting anti-cancer, anti-inflammatory, and anti-diabetic properties,
and potential usage in traditional medicines (Collins et al. 2022
). The release of
medicinally prominent SMs from both these plants are predominantly influenced by
environmental stresses. During cold stress overexpression of CBFs (mainly CBF1)
promote the activity of catabolic effectors i.e., DELLA RGL3 and anabolic
inhibitors like GA2oxy resulting in the increased hydroxylation and inactivation of
the bioactive GA. Such an inactivation in GA establishes ross-talk with other
hormones like Auxins, Jasmonic acid and Brassinoste
roids to exert their growth-
promoting signals (Eremina et al. 2016).
182 G. K. Dugganaboyana et al.
Methyl Jasmonates
Jasmon
ates are bioactive lipid soluble hormones derived from linolenic acid via the
COX, LOX, and Cytochrome p450 pathways. The biosynthesis of jasmonic acids is
widely influenced by ambient temperature and other stress conditions. Under cold
stress, oxygenase-mediated JA biosynthesis from PUFA is greatly disturbed and
further genesis is ceased. As a result, during cold acclimation JA tackles the cold
injury by promoting the overexpression of certain COR genes that translates into
effector signaling molecules that could rewire/repair the detrimental effects of cold
injury in plants (Zhao et al.
2005). The primary responses are the accumulation of
antioxidant enzyme systems that could minimize the oxidative damage and protects
the membrane homeostasis. Jasmonate Zim Domain (JZD) i.e., JZD1 and JZD2 in
conjugation with the repressors ICE1 and ICE2 repress or cease the expression of JA
responsive genes and also influence the CBF activity. Inactivation of this complex
and abundant biogenesis of JA is another downstream signal transduction mecha-
nism elicited in response to cold stress.
Gymnema sylvestre (Aka. Gurmar), a reputed herb in Ayurveda is widely known
for its SMs like triterpene saponins, gymnemic acid, gurmarin, etc. all of which are
envoys in suppressing sugars in the blood and maintain sugar homeostasis. The herb
also evinces several curative properties against anemia, osteoporosis, infections,
bowel disorders, cholesterol, lipid storage disorders, etc. Activation of Jasmonate
responsive genes during cold stress consequently triggers the enzymes needed for
the biogenesis of gymnemic acid along with several other SMs like alkaloids,
flavonoids, phenylpropanoids, etc. and acclimate the herbal plant to cold stress and
protect it from the repercu ssions of cold injuries (Chodisetti et al.
2015).
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11 Signaling Molecules in Medicinal Plants Response to Cold Stress 183
Salicylic Acid
Ubiquitously distrib uted SA has comprehensive roles in regulating the development,
growth, pathogen infection, and defense mechanisms against abiotic and biotic stress
signals. It is one of the components of Choristmate that accumulate during cold
stress and results in hypersensitive response (HR). Primarily, SA induces free
radicals or ROS in response to chilling or freezing temperatures. ROS further
activate the antioxidant and non-antioxidant enzyme systems promoting the cold
acclimation in plants. Large-scale omics studies have been well established to
determine changes in the expression and accumulation of SA in the Genus Jasminum
grown in cold temperatures. Jasminum offcinale, pharmacologically active herb
releases several SMs like anthocyanins, saponins, steroids, emodine, terpenes,
tannins, coumarins, flavonoids, etc. with certain antimicrobial, antioxidant, insecti-
cidal and dermatologic benefits (Al-Snafi
2018). Jatropha curcas L. and
Andrographis paniculate are amongst other herbal plants in which SA acid under
cold stress conditions promote a dramatic increase in MDA leading to electrolyte
leakage and oxidative stress. This greatly disrupts the biogenesis and accumulation
of principal SMs. The accumulated MDA elicits the antioxidant systems and in turn
also increases the photosynthetic rates in these plants leading to cold accli mation.
Further, the cross-talk between the SA and GA ceases plant growth and development
in during cold injury and prioritizes the activation of repair and stress tolerant
mechanisms during cold acclimation (Eremina et al. 2016).
Ethylene
Ethylene is a multi-functional phytohormone synthesized from S-adenosyl-
L-Methi-
onine to form aminocyclopropane-1-carboxylic acid (ACC), a rate-limiting step
catalyzed by ACC synthase. The role of ethylene in plants is a balanced contrast
i.e., based on the ambient conditions, time of application and/or diversity in the
species ethylene either promote growth or foster senescence. Unlike the other plant
hormones, the role of ethylene during cold stress is often debated as it is species
specific and not certain.
Medicago sativa (Aka. Alfalfa), deep-rooted leguminous plant o f Fabaceae
family is widely used in traditional medicine for its phytochemical constitution.
Isoflavanoids, flavones, phenolic acids, tannins, alkaloids, terpenes, carotenoids,
phytoestrogens, and glycosides are some of the prominent SMs effective against
infections, CVDs, diabetes, oxidative stress, liver diseases, Rheumatoid arthritis,
neurodegenerative diseases and so on (Bora and Sharma
2011). On the other hand,
Rheum australe (Aka. Himalayan Rhubarb), an endangered Himalayan medicinal
herb propertied with diverse SMs like stilbenes, flavonoids, anthraquinones, etc. are
well known for numerous ethnomedical applications for the treatment of arthritis,
gastritis, liver disorders, bronchitis, dysentery, flatulence, scabies, swelling and so on
(Pandith et al.
2018). In both these cases the expression levels of SAM synthase
2 (SAM2) and ACC oxidase is upregulated and facilitates the increased biogenesis
of ethylene. Due to the cold stress, it is understood that, increase in ethylene
terminate the growth of the plant and initiate the expression of several COR genes
and species-specific TFs like CBF1–3/DREB which in turn initiate the biogenesis of
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several enzymatic systems (specifically, peroxidase, superoxide dismutase, and
catalase) and osmolytes to fight the oxidative damage and increase freeze tolerance
(Echeverria et al. 2021).
184 G. K. Dugganaboyana et al.
The negative regulation of
cold stress by ethylene is more predominant than its
positive defense mechanisms. Ethylene-arbitrated hypersensitivity to cold stress in
Medicago truncatula is one of the notable examples that evince contrasting and
species-specific roles of ethylene during cold stress. The expression of MtCBF1,
MtCAS15 and MtCBF3, (Cold Acclimation Specific genes) is greatly compromised
by a negative signal transduction pathway. Ethylene due to cold stress binds to ETR1
(Ethylene Responsive gene 1) and activates CTR1 (Constitutive Triple Response 1)
which phosphorylates the downstream transcription factors like EIN2 (ethylene
insensitive 2) which further upregulate EIN3 and ein3-like 1 (EIL1). These tran-
scription factors suppress CBFs expression leading to the hypersensitivity of plants
to cold stress (Eremina et al. 2016).
11.2.1.6 Secondary Metabolites: Prerequisites for the Sanative Actions
of Medicinal Plants
Recent studies estimated roughly about 80% of the global population has switched
from synthetic drugs to herbal therapies as they are natural nutraceutical products
that succor to overcome ailments and promote healthy aging (Pant et al. 2021).
Secondary plant metabolites (SMs) are the conspicuous bioactive metabolic
by-products or signaling molecules derived from the primary catabolic pathways
in the plant cells. Polyphenolics, alkaloids, saponins, terpenes, coumarins,
flavonoids, lipids, carbohydrates, etc. derived from different plant parts (root,
stem, leaves, flowers, fruits, and seeds) have been procl aimed to underpin various
pharmacological benefits in herbal plants (Hussein and El-Anssary 2018). In addi-
tion to the therapeutic properties, SMs are of equal prominence in plant defense
mechanisms. Therefore, the biogen esis, distribution, and accumulation of these
bioactive molecules rely on several biotic and abiotic stresses owning to herbal
predators and environmental influences. The Intergovernmental Panel on Climate
Change (IPCC) has announced that temperature variations have a significant impact
on the SM profile in medicinal plants resulting in their amendment. Previous sections
address certain striking effects of cold on the biogenesis of certain pharmaco-
logically pronounced SMs and underlying cold acclimation/tolerance mechanisms
adopted by medicinal plants during cold stress and their repercussions on their
therapeutic value. It is essential to understand the role of certain SMs themselves
in achieving cold acclimation/tolerance.
Brassica rapa (Aka. Turnip) is one of the classic medicinal herbs known to have
curative effects on several diseases that are part-and-parcel of human life.
2-phenylethyl isothiocyanate, phenyl propionitrile, brassica phenanthrene A,
flavonoids, and phenolics are the most prominent SMs with pharmacopeial
investigations in the treatment of headaches, angina, rheumatisms, STDs, hyperten-
sion, diabetes, neurodegenerative diseases, brain disorders and so forth (Paul et al.
2019). The phenolic extracts of this plant are widely known to possess anti-oxidant
mechanisms that facilitate in cold accli mation. O-glycoside, prominent flavanol in
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Role during cold acclimation Reference
turnip govern the growth and development of radical and plumule during seed
germination. Chilling and freezing temperature induced physiological alterations
dormant the seed growth leading to their death. The indigenous gene makeup in
these plants enable the expression of COR genes that encode several SMs predomi-
nantly, polyphenols and flavonoids that promote several carbohydrate anabolic
pathways, protein folding in the ribosomes, activation of the photosynt
hetic systems
in chloroplast, upregulation of antioxidant enzymes, and others. Facilitating the plant
to get away from cold-induced injuries (Ilyas et al.
2022).
11 Signaling Molecules in Medicinal Plants Response to Cold Stress 185
Table 11.1 Concerted role of diverse secondary metabolites released in Rhodiola semenovii under
cold stress
Secondary
metabolite
Site of
biosynthesis
β, γ- Sitosterol Roots Integral component of the plasma membrane.
Prevents membrane-crystallization and
promotes fluidity
Stabilizes the damage of the thylakoid
membrane and magnifies the chlorophyll
synthesis
Shahzad
et
al. (
2021)
γ-Tocopherol Shoot Natural antioxidants that form phenoxy
radicals in conjugation with hydrogen
peroxide and transform into quinones, dimers,
and trimers
Rontani
et al. (
2007)
Linoleic and
linolenic fatty
acids
Shoot Intrinsic to plasma membrane these PUFAs
help in regulating micro viscosity,
permeability of electrolytes, deaden
excitability, and activate enzymes that are
responsible for the biosynthesis of membrane
proteins and lipids
Terletskaya
et al. (
2021)
Oxylipins Shoot and
root
Oxylipins are complex interactive molecules
that establishes a cross-talk with various
phytohormones like SA, ET, IAA, BA, GA,
ABA, etc. and bring about a cascade of
downstream hormonal interactions and
promote tolerance in plants
Pushkina
(
2020)
Phenols Shoot and
root
Prevents mitochondrial autolysis, promote
membrane stability by inhibiting/blocking the
ROS generation
Pushkina
(
2020)
Terpenes Shoot Diterpene like phytol promote chlorophyll
synthesis and triterpene like squalene are
scavengers of ROS, protectants of the LHC
from photoinhibition
Edge et al.
(
1997)
Rhodiola semenovii Boriss. is a succulent whose concoctions have been used
widely as traditional medicine in Central Asia for treating inflammatory diseases,
diabetes, cancer and many others (Terletskaya et al. 2021). It is famed for its
mult
imode acclimation strategies against abiotic stresses with rich and diversified
SMs (Table
11.1).
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186 G. K. Dugganaboyana et al.
Ocimum basilicum L. hoard a hub of mono and sesquiterpenes especially during
cold stress. At the onset of cold injury, genes encoding various terpenoids like
linalool synthase (LIS), β-myrcene synthase (MYS), geraniol synthase (GES),
germacrene D synthase (GDS), Farnesyl diphosphatase, and γ-cadinene synthase
(CDS) expression levels show a prepotent expression leading to their increased
biosynthesis. This indicates that the cold responsive genes (COR) are present within
the promoter region of the terpenoids. These compounds therefore help the plant
battle the effects of cold injury and attain acclimation or tolerance (Senji and
Mandoulakani 2018).
11.3 Conclusion
The role of medicinal plants and the efficacy of their critical metabolites in the
deterrence of diverse illnesses have been a prime focus of research since time
immemorial. The seed, fruit, tuber, root, leaf, and bark extracts housing enormous
phytochemicals have been conceded in the healing science for its holistic, natural,
and safe curative strategies. Attempt has been made globally for a substantial
cultivation of these traditional herbs, recognize the salient phytochemicals with
therapeutic properties, develop formulations, and proffer to the expanding popula-
tion. Climate change and increasing biotic and abiotic stresses primarily transcend
the overall yield and phytochemical constitution of these medicinal plants. To
prevail over these damages especially, the chilling and freezing injuries, plants
have evolved numerous defense mechanisms governed by a troop of signalling
molecules and downstream signal transduction cascades. In this chapter, attempt
has been made to understand various cold acclimation strategies adopted by medici-
nal plants to protect themselves from the desecration of cold stress. Consorted
physiological, morphological, and biochemical alterations in plants due to cold
injury evoke the expression of several COR genes and associated transcription
factors like CBF/DREB leading to the activation and accumulation of several
cryoprotectants in various parts of the plant. In addition to the primary cold-induced
injury, secondary oxidative and osmotic damage also contribute to the death of the
plants. Several Antioxidant and non-antioxidant mechanisms including enzymes,
proteins and vital phytohormones are functionally activated in plants to combat the
oxidative damage and protect the membrane integrity. Likewise, severa l sugars,
amino acids, and amines act as cryoprotectants to maintain the osmo tic homeostasis.
Secondary metabolites formed during the primary catabolic pathways play a critical
role in altering the gene expression pattern, establish cross-talk amongst the
phytohormones, ensure membrane stability, scaveng e the generated free-radicals,
promote photosynthesis, prevent dehydration , and thereby protect the plants from
cold-induced damages and death. In the recent times, engineering the COR genes
and associated cryoprotective proteins in transgenic plants have improved the cold
tolerance mechanisms and promote the biogenesis of clinically eminent secondary
metabolites.
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11 Signaling Molecules in Medicinal Plants Response to Cold Stress 187
Acknowledgement The authors are sincerely thanks to the authorities of JSS Academy of Higher
Education and Research, Mysuru, India for infrastructural
and computational facilities.
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