J Plant Growth Regul (2013) 32:216–232
DOI 10.1007/s00344-012-9272-x
Role of Secondary Metabolites and Brassinosteroids in Plant
Defense Against Environmental Stresses
Arti Bartwal • Rakesh Mall • Pushpa Lohani •
S. K. Guru • Sandeep Arora
Received: 10 June 2011 / Accepted: 5 March 2012 / Published online: 13 May 2012
 Springer Science+Business Media, LLC 2012
Abstract Being sessile, plants are subjected to a diverse
array of environmental stresses during their life span.
Exposure of plants to environmental stresses results in the
generation of reactive oxygen species (ROS). These acti-
vated oxygen species tend to oxidize various cellular bio-
molecules like proteins, nucleic acids, and lipids, a process
that challenges the core existence of the cell. To prevent
the accumulation of these ROS and to sustain their own
survival, plants have developed an intricate antioxidative
defence system. The antioxidative defence system com-
prises various enzymatic and nonenzymatic molecules,
produced to counter the adverse effect of environmental
stresses. A sizable number of these molecules belong to the
category of compounds called secondary metabolites.
Secondary metabolites are organic compounds that are not
directly involved in the growth and development of plants
but perform specialized functions under a given set of
conditions. Absence of secondary metabolites results in
long-term impairment of the plant’s survivability. Such
compounds generally include pigments, phenolics, and so
on. Plant phenolic compounds such as flavonoids and
A. Bartwal
R. Mall
Department of Biochemistry, G. B. Pant University of
Agriculture & Technology, Pantnagar, Uttarakhand,
India 263145
P. Lohani
S. Arora (&)
Department of Molecular Biology and Genetic Engineering,
G. B. Pant University of Agriculture & Technology, Pantnagar,
Uttarakhand, India 263145
e-mail: plantstress@gmail.com
S. K. Guru
Department of Plant Physiology, G. B. Pant University of
Agriculture & Technology, Pantnagar, Uttarakhand,
India 263145
123
lignin precursors have been reported to accumulate in
response to various biotic and abiotic stresses and are
regarded as crucial defence compounds that can scavenge
harmful ROS. Another important category of plant
metabolites, called brassinosteroids, exhibit stress regula-
tory and growth-promoting activity and are classified as
phytohormones. Elucidation of the physiological and
molecular effects of secondary metabolites and brassinos-
teroids have catapulted them as highly promising and
environment-friendly natural substances, suitable for wider
application in plant protection and crop yield promotion.
The present review focuses on our current understanding of
how plants respond to the generation of excessive ROS and
the role of secondary metabolites and brassinosteroids in
countering the adverse effects of environmental stresses.
Keywords Environmental stress
Reactive oxygen
species
Antioxidative defence
Secondary metabolites

Brassinosteroids
Introduction
During the course of development, plants are exposed to a
variety of environmental stresses that alter the basic
metabolism of the affected plants. To successfully com-
plete their life cycle, plants must cope with environmental
constraints that include pathogen attack, extreme temper-
ature fluctuations, water deficit, and high light intensity.
When these parameters exceed a critical limit, plants are
said to be under stress. This shifting of environmental
conditions then leads to reduced growth rate and lower
productivity, and, in extreme circumstances, may even
prove fatal. Environmental stresses cause losses worth
billions of dollars in crop yield, affecting the magnitude of
J Plant Growth Regul (2013) 32:216–232
productivity on a year-to-year basis (Apel and Hirt 2004).
Further, beyond the single-plant level, fluctuations in
environmental conditions can also select for short- and
long-term shifts in ecological traits, affecting ecosystem
functioning, carbon sequestration, and biological diversity.
Environmental stresses are divided into two major
groups: biotic and abiotic. Biotic stresses include various
pathogenic microorganisms, weeds, fungi, and other pre-
dators. Sources of abiotic stress include air pollutants,
extreme temperatures (including freezing), drought, high
light intensity, salinity, and mechanical damage (Fig. 1)
(Vickers and others 2009). Rapidly rising temperatures,
drought, and pollutants are the most salient phenomena
associated with global climate change. This unprecedented
climate change now threatens the existence of plants.
Physiological and molecular changes at the cellular level
that are associated with various abiotic stresses include
turgor loss, changes in membrane fluidity and composition,
changes in solute concentration, and protein-protein and
protein-lipid interactions (Chaves and others 2003).
Reduction in photosynthetic activity, accumulation of
organic acids and specific osmolytes, and changes in car-
bohydrate metabolism are typical responses to various
abiotic stresses. The reduction in photosynthetic activity is
a consequence of numerous coordinated events such as
stomatal closure, inefficient electron transport, and reduced
activity of photosynthetic enzymes.
Fig. 1 Plant subjected to
various environmental stresses
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Very few plant responses are stress-specific and most of
the stresses elicit generic responses like production of
excessive reactive oxygen species (ROS) (Apel and Hirt
2004). One of the primary responses of plants exposed to
drought, salinity, or osmotic stresses is the synthesis of
osmoprotectants, also known as compatible solutes. These
osmoprotectant molecules accumulate in plants exposed to
different environmental stresses and help maintain cellular
homeostasis and detoxification of ROS (Tafaeizadeh 1998;
Rontein and others 2002). These molecules include amino
acids, polyols, quaternary ammonium, and tertiary sulfo-
nium compounds. The cellular balance of ROS is normally
kept under tight control. However, when this dynamic
control is lost, consequential damage occurs. ROS cause
direct damage to plant cells through oxidation of biological
components like nucleic acids, proteins, and lipids. Plants
have developed an intricate defence response network of
lipophilic and hydrophilic antioxidant compounds and
enzymes that provide protection against conditions of
excessive oxidative damage. Direct defence reactions
quench and remove ROS whereas indirect responses
include hormone-mediated signalling to upregulate an
array of stress defensive genes (Kwak and others 2006).
Measuring abiotic stress responses and attributing mecha-
nistic behaviour can be cumbersome because of the com-
plexity of the ROS response network, which makes it
difficult to distinguish the cause-and-effect relationships.
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Fig. 2 Hydroxyl radical-
mediated peroxidation of
linolenic acid in lipids
The defence network is linked intricately between lipo-
philic and hydrophilic phases. The dynamic balance
between the accumulation of ROS under stress and its
detoxification by the cellular antioxidative defence
machinery holds the key to the stress tolerance attributes of
a plant.
Generation of ROS and Associated Regulatory
Mechanisms
The oxidation of water by the photosystem-II complex
results in the production of molecular oxygen. However,
oxygen can also act as a potential electron acceptor,
resulting in the formation of superoxide radicals. Super-
oxide radicals must be eliminated immediately by the
antioxidative defense system. In addition, significant
amounts of hydrogen peroxide (H2O2) are also produced in
cells because of various metabolic reactions. Although
much of this H2O2 is catabolized by catalase and peroxi-
dases, some chemical decarboxylation of keto acids by
H2O2 occurs. ROS formation is initiated by the univalent
reduction of molecular oxygen using either one, two, or
three electrons generating superoxide, hydrogen peroxide,
or hydroxyl radical, respectively, or by the formation of
singlet oxygen by transfer of excess excitation energy to
molecular oxygen. ROS are divided into two main classes
consisting of nonradical species (H2O2) and free radical
forms. Accumulation of higher levels of ROS is potentially
detrimental to plant cells, causing damage to critical bio-
molecules like DNA, proteins, lipids, chlorophyll pig-
ments, and membrane components (Jaleel and others
2009). Oxidation of organic substrates may proceed by two
possible reactions: (1) addition of OH• to an organic
molecule or (2) removal of a hydrogen atom from organic
molecules. The addition of OH• to organic substrate forms
a hydroxylated product that is converted to a stable
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J Plant Growth Regul (2013) 32:216–232
oxidized product on further oxidation by Fe3?, O2, or other
agents. The dismutation of hydroxylated product can lead
to the formation of crosslinked products. In the removal
reaction, the OH• radical oxidizes the organic substrate by
forming water and an organic radical. The organic radical
has a single unpaired electron and thus can react with
oxygen in the triplet ground state. The addition of triplet
oxygen to the organic radical leads to the formation of a
peroxyl radical, which can readily remove hydrogen from
another organic molecule leading to the formation of a
second organic radical. The hydrogen removal reaction of
the hydroxyl radical is best demonstrated by peroxidation
of linolenic acid in cell membranes. The lipid peroxides
(ROOH) are highly unstable in the presence of Fe2? or
other reduced metal ions (such as Cu?), as they participate
in the Fenton reaction leading to the formation of a reactive
alkoxy radical. The alkoxy radical so formed is as dam-
aging as the hydroxyl radical and starts a cascade of oxi-
dative reactions (Figs. 2, 3).
The scavenging of O•2 - (superoxide radical) by super-
oxide dismutase (SOD) results in the production of H2O2,
which is removed by ascorbate peroxidase (APX) or cat-
alase (CAT). However, both O•2 - and H2O2 are not as
toxic as the OH• radical, which is formed by the combi-
nation of O•2 - and H2O2 in the presence of trace amounts
of Fe2? and Fe3? by the Haber–Weiss reaction. The OH•
can potentially damage all major biological macromole-
cules, including chlorophyll, protein, DNA, and lipid
molecules, thus severely affecting plant metabolism and
limiting growth and yield. In the acute condition, it may
also lead to the death of the plant. To combat such ROS-
mediated oxidative damage, the plant employs a series of
enzymatic and nonenzymatic detoxification systems, thus
providing itself protection against oxidative damage
(Sairam 1994). For example, overexpression of SOD in
plants affects a number of physiological phenomena,
J Plant Growth Regul (2013) 32:216–232
Fig. 3 Generation and
scavenging of superoxide
radical, hydrogen peroxide,
hydroxyl-induced lipid
peroxidation, and glutathione
peroxidase-mediated fatty acid
stabilization under
environmental stresses
including the metabolism of H2O2, biosynthesis of lignin in
cell walls, auxin catabolism, generic defence responses to
wounding and pathogen or insect attack, and specific
alterations in respiratory processes. The incidence of
expression and activation of APX is directly related to the
appearance of physiological injuries caused in plants by
oxidative stress. A decrease in antioxidant capacity in
stressed tissues results in higher levels of ROS that may
contribute to further injury. Consequently, protection
against oxidative stress is an important component in
determining the survival of a plant under stress. Studies on
heat-acclimated versus nonacclimated cool season turf
grass species suggest that the former had lower production
of ROS due to enhanced synthesis of ascorbate and glu-
tathione (Xu and others 2006).
Secondary Metabolites
Secondary metabolites, also referred to as secondary prod-
ucts or natural products, are organic compounds that are not
directly involved in growth, development, or reproduction.
They do not even directly participate in respiration, trans-
location, protein synthesis, nutrient assimilation, photosyn-
thesis, or differentiation. However, the absence of secondary
metabolites may result not only in long-term impairment of
the plant’s survivability but also in immediate death. These
compounds tend to be more complex than primary metabo-
lites. Such compounds generally include pigments, antitu-
mor agents, effectors of ecological competition and
symbiosis and molecules of plant chemical defence such as
alkaloids and steroids. Although primary metabolites are
219
present in every living dividing cell, secondary metabolites
are found occasionally and are not of much significance for
primary plant life. Secondary metabolites are generally
present at low concentrations in all living organisms; how-
ever, these compounds are widely distributed in plants and
play a crucial role. Recently, such compounds have become
the subject of increasing interest because of their significant
practical use for nutritive, medicinal, and cosmetic purposes,
as well as their indisputable role in plant stress physiology.
Because plants are devoid of motility and an immune system,
they have developed alternative defence strategies that
involve a vast variety of secondary metabolites serving as
tools to overcome stress constraints, adapt to the changing
environment, and thus help in survival under suboptimal
conditions (Edreva and others 2008). There is a delicate
balance between generation and destruction of oxidant
agents, which may be beneficial or deleterious to the plants.
Under physiological conditions, the level of ROS is kept low
by the coordinated activity of antioxidative systems that
include secondary plant metabolites and scavenging
enzymes. Many in vitro studies have found that compounds
like sesquiterpenes, flavonoids, coumarins, and phenolic
acids have substantial antioxidant activity; for example,
flavonoids have been studied extensively for their antioxi-
dant activities (Khan and others 2010). Transcriptional reg-
ulation has long been recognized as a major determinant of
phenotypic variation (Wray and others 2003). Using classi-
cal molecular biology techniques, coordinated expression of
secondary metabolite synthesizing enzymes in Arabidopsis
has been described in several reports, for example, in stress-
inducible indole metabolism (Zhao and others 1998) and
in flavonoid synthesis during seedling development
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(Pelletier and others 1999). Likewise, key enzymes of
the phenylpropanoid pathway, namely, phenylalanine
ammonia-lyase (PAL), coumarate-4-hydroxylase (C4H), and
4-coumaroyl CoA ligase (4CL) isoforms, are expressed in a
coordinated manner under stress conditions (Koopmann and
others 1999). However, results of earlier studies on secondary
metabolism need to be reinterpreted in light of abundant
duplication of the underlying genes (Bowers and others 2003).
Several different secondary metabolites have been
identified in the approximately 500,000 higher plant spe-
cies found on our planet. As the name indicates, secondary
metabolites were considered the side products of plant
metabolism. This attitude reflected a lack of knowledge in
the scientific community, ignoring the fact that cellular
metabolism is an intricate result of the interaction of a plant
with its environment.
Categories of Secondary Metabolites
There are three major groups of secondary metabolites
(Michalak 2006):
(1) Terpenes (terpenoids or isoprenoids):
• 2 isoprene units (C10 terpenes) are termed monoterpenes
• 3 isoprene units (C15 terpenes) are termed sesquiterpenes
• 4 isoprene units (C20 terpenes) are termed diterpenes
• 6 isoprene units (C30 terpenes) are termed triterpenes
• 8 isoprene units (C40 terpenes) are termed tetraterpenes
• 8 isoprene units ([C40 terpenes) are termed
polyterpenoids
(2) Phenolics: flavonoids, tannins, lignins, and so on.
(3) Nitrogen-containing secondary metabolites: alkaloids
such as cocaine, caffeine, and morphine; cyanogenic
glycosides and glucosinolates, and so on.
Properties of Secondary Metabolites
Diversity in Chemical Nature
More than 100,000 different secondary metabolites have
been reported (Hadacek 2002). The most characteristic
feature of secondary metabolites is the large diversity of
chemical types, representative of all the main classes of
organic compounds: aliphatic, aromatic, hydroaromatic,
and heterocyclic. Unique carbon skeletons occur along
with a multiplicity of functional groups.
Characteristics and Functions
Secondary metabolites perform a broad array of protective
functions, including antimicrobial, photoprotective, structure-
stabilizing, and signalling, under biotic and abiotic stress sit-
uations. Their diverse chemical characters can be the
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determinants of various functions. Several secondary metab-
olites are synthesized in plants from the intermediates of pri-
mary carbon metabolism via phenylpropanoic acid, shikimic
acid, mevalonic acid, and methyl erythritol phosphate path-
ways (Taiz and Zeiger 2006). Abiotic stresses induce pro-
duction of various secondary metabolites like flavonoids and
other phenylpropanoids. The increased activity of PAL in
response to stress is one of the main acclimatory responses.
Research has shown that lipids of the thylakoid membrane are
stabilized and photoprotected by various carotenoids of the
xanthophyll family and some terpenoids such as isoprene and
a-tocopherol. The xanthophyll cycle also plays a pivotal role
in photoprotection of plants. Carotenoids are reported to have
a role in protecting cellular structures in several plant species
under different stresses (Havaux and Gruszecki 1993; Havaux
1998). Phenolics, including flavonoids, lignin, and anthocy-
anins, play a variety of roles in plants as they are the most
abundant class of secondary metabolites. Phenolics have been
reported to accumulate under extreme stress conditions,
accompanied by increased concentrations of PAL but
decreased activities of peroxidase polyphenyl lyase (Sgherri
and others 2004; Taiz and Zeiger 2006). The level of flavo-
noids is modulated in plant tissues under stress conditions
(Sachray and others 2002). High temperature reportedly
decreases the synthesis of anthocyanins in Chrysanthemum
(Shibata and others 1988) and aster and red apple (Tomana
and Yamada 1988). Emission of some volatile compounds
belonging to the family of isoprenoids has been shown to
confer abiotic stress tolerance in plants. Isoprene production
protects photosystem II from the damaging effects of ROS
(Taiz and Zeiger 2006). Its endogenous production protects
the biological membranes by directly binding with singlet
oxygen by means of an isoprene conjugate double bond
(Velikova and others 2004).
Water solubility contributes to the compatible secondary
metabolites involved in osmoregulation and protection of
hydrophilic cellular sites under stress, whereas hydrophobic
molecules (carotenoids) are the best candidates to protect
lipophilic surfaces such as membranes. It is also becoming
increasingly evident that secondary metabolites play an
important role as signalling molecules. However, the chemi-
cal rationale of their involvement in the signal transduction
cascades remains to be fully elucidated (Edreva and others
2008). Colour and scent production by secondary metabolites
can attract or repel insects and herbivores, whereas toxins can
be involved in plant-plant allelopathic interactions thus pro-
tecting plants from biotic stress (Hadacek 2002).
Role of Electrical Charge and Important Functional
Groups
Electrostatic interactions of secondary metabolites with
other biomolecules can result in stabilizing cell structures.
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The protonated amino and imino groups in polyamines
have a positive charge that allows electrostatic interactions
with negatively charged groups in macromolecules and
cellular substructures, providing a stabilizing effect.
Reports suggest that electrostatic interactions of poly-
amines with phosphoric acid residues in DNA, uronic acid
residues in the cell wall matrix, and negative groups on
membrane surfaces help maintain their functional and
structural integrity (Edreva and others 2007).
The covalent bonding of functional groups brings about
polymerization, condensation, and complexation events
that occur mainly at the cell wall level. Thus, polymeri-
zation of phenolic alcohols (sinapyl-, coniferyl-, and p-
coumaryl-) yield lignins, which cause strengthening of the
wall structure. The association of the polysaccharide chain
in the cell wall matrix and ferulic acid (an unsaturated
aromatic carbonic acid) is brought about by the formation
of an ester bond. Crosslinking via diferulate bridges also
consolidates the cell wall structure (Fry 1986). The pres-
ence of O-diphenol groups allows formation of O-quinoid
grouping. In vitro covalent interactions between proteins
and oxidation products of caffeoylquinic acid have recently
been reported (Prigent and others 2007). Quinoid grouping
can interact with thiol and amino groups in proteins as
follows:
Incidentally, the toxicity of secondary metabolites due
to the quinoid group interactions with microbial proteins
provides resistance against pathogenic fungi, bacteria, and
viruses, thus underlining the antimicrobial activity of these
compounds. These interactions block specific active sites
and the corresponding function of proteins. Similar inter-
actions of quinoid groupings with plant proteins can be a
possible mechanism for the necrotic reaction against plant
pathogens. High molecular condensation products of the
quinoid form of O-diphenol and protein were reported to
arise during necrosis and could limit the spread of stress-
induced tissue damage (Edreva and others 2008).
The photoprotective function of various secondary
metabolites (flavonoids, including anthocyanins, cinnamic
acid derivatives, and xanthophylls) can be explained by the
presence of conjugated double bonds, that is, delocalized
p-electrons. Due to this electronic configuration, they tend
to absorb visible and UV radiation, thereby causing easy
221
electron and energy transfers. p-electron configuration
occurs in closed-ring, short side chain, and long-chain
structures (Demmig-Adams 2003).
An increasing amount of evidence has shown that sec-
ondary metabolites function as antioxidants and antiradi-
cals, helping the plants to cope with oxidative stress that
arises in hostile environments (Gould and others 2002).
The growing list includes hydroxy and thiol group-con-
taining compounds such as ascorbic acid and lipoic acid,
O-dihydroxy group-containing flavonoids such as querce-
tin, aliphatic and arylamines, unsaturated fatty acids, and
carotenoids (Edreva and others 2007). Chelation of tran-
sition metals by flavonoids such as quercetin interferes
with the generation of ROS via the Fenton reaction, thus
contributing to a powerful antioxidant/antiradical perfor-
mance (Edreva and others 2008). The chemical types of
some of the secondary metabolites with their chemical
formulae are given in Table 1, and their chemical charac-
teristics with respect to their interaction and function are
given in Table 2.
Terpenes as Protective Secondary Metabolites
Terpenes represent the largest group of plant secondary
metabolites. Ten thousand of terpenes have been isolated
and purified and their structures elucidated. They are
widespread in nature and occur in almost all plants (Obst
1998). Terpenes are hydrocarbons derived from isoprene
(isopentane) C5 units. They perform several functions in
plants; for example, they act as plant growth regulators,
defense molecules against herbivores and pathogens,
attracting compounds for pollinators, and as compounds
that influence (directly or indirectly) the growth and
development of neighbouring plants. Until recently, the
function of terpene emissions from some plants that do not
store terpenes in their tissues has been a mystery. Fol-
lowing the report that isoprene, a hemiterpene, increases
thermotolerance and photosynthesis in some species
(Sharkey and Singsaas 1995; Loreto and others 1998), it
was discovered that thermotolerance of the monoterpene-
emitting oak (Quercus ilex) increases when leaves are
fumigated with monoterpenes. Terpenes play a role in
thermotolerance, suggesting that the mechanism by which
this occurs is not limited to plants that synthesize these
compounds. Terpene storage allows plants to maintain
sufficient quantities to be detected by animals (in the case
of attracting pollinators) or to be released upon tissue
damage (in the case of defence against herbivores). Thus,
interaction with insects typically requires storage of terp-
enes within the plant tissues. Certain terpenes have a well-
characterized role in plant growth and development (Khan
and Mohammad 2011). Sterols are terpene derivatives that
are essential components of the cell membrane, which they
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Table 1 Different categories of secondary metabolites
Chemical types Formulae
Aliphatic
Aromatic
Hydroaromatic
Heterocyclic
stabilize by interacting with phospholipids. The red,
orange, and yellow carotenoids are tetraterpenes that
function as accessory pigments in photosynthesis and
protect photosynthetic tissues from photooxidation. The
hormone abscisic acid is a C15 terpene produced by deg-
radation of a carotenoid precursor (Taiz and Zeiger 2006).
Carotenoids fulfill two major roles in photosynthetic
organisms. Their first role is to act as light-harvesting
pigments, extending the range of the light spectrum
available for use in the photosynthetic process. They
absorb light in the range of 450-570 nm, whereas chloro-
phyll molecules do not, and they pass on the captured
energy to the chlorophylls. Secondly, carotenoids provide
photoprotection to the photosynthetic system (Price and
others 1989). Carotenoids also prevent the formation of
singlet oxygen by quenching the triplet state of the chlo-
rophyll molecules as they arise (Fyfe and others 1995).
Phenolics as Antistress Secondary Metabolites
In plants, phenolics such as flavonols and phenylpropa-
noids act as potential antioxidant compounds by donating
electrons to guaiacol peroxidases (GuPXs) for detoxifica-
tion of high amounts of H2O2 produced under stress
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Representatives
Polyamines
Ethylene
Phenolic alcohols
Phenolic acids
Unsaturated aromatic
carbonic acids
Jasmonic acid
Flavonoids
conditions. The antioxidant/pro-oxidant activity of phyto-
phenolics depend on their metal-reducing potential, che-
lating behaviour, pH, and solubility (Decker 1997). Most
plants constitutively synthesize low concentrations of
phenylpropanoids such as flavonoids and hydroxycinna-
moyls (HCAs). However, accumulation of phenolics in
plants can be induced by abiotic and biotic stresses, for
example, UV radiation, high light intensity, low tempera-
ture, wounding, low nutrients, and pathogen attack
(Yamasaki and others 1995). It has recently been shown
that flavonoids and HCAs are capable of scavenging H2O2
by acting as electron donors for GuPXs such as horseradish
peroxidase (HRP) (Sakihama and others 2000). Polyphe-
nols, particularly flavonoids and tannins, have long been
associated with plant defence against herbivores. An
increase in phenolic content has also been observed in
poplar and oak trees in response to feeding and wounding
(Hammerschmidt and Schultz 1996). Oxidation of pheno-
lics usually occurs because of tissue damage. The reactive
species responsible for the DNA nicking activity of phen-
olics is the hydroxyl radical or a species with similar oxi-
dative potential (Li and Trush 1994).
The reduced forms of phytophenolics are powerful
antioxidants equivalent to ascorbate. In contrast, the
J Plant Growth Regul (2013) 32:216–232
Table 2 Biological functions of
specific secondary metabolites
phenoxyl radical produced through antioxidative reactions
and in lignin biosynthesis is a potential pro-oxidant. Phen-
olics are characterized by at least one aromatic ring (C6)
bearing one or more hydroxyl groups. These are synthesized
mainly from cinnamic acid, which is formed from phenyl-
alanine by the action of L-PAL, the branch-point enzyme
between primary (shikimate pathway) and secondary
(phenylopropanoid) metabolism. The hydroxyl groups of
phenolic compounds are able to bind iron and copper (Jung
223
and others 2003). They may inactivate iron ions by che-
lating and additionally suppressing the superoxide-driven
Fenton reaction, which is believed to be the most important
source of ROS (Rice-Evans and others 1997). The induction
of phenolic compound biosynthesis was reported in wheat
in response to nickel toxicity and in maize in response to
aluminium (Winkel-Shirley 2002). It was found that when
Phaseolus vulgaris is exposed to Cd2?, it accumulates a
significant amount of phenolics. It was also reported that
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Phyllantus tenellus leaves contained more phenolics than
control plants after being sprayed with copper sulphate
(Bors and others 1990). The concept of antioxidant action of
phenolic compounds is not novel. There have been many
reports of induced accumulation of phenolic compounds
and peroxidase activity in plants under stress. The roots of
many plants exposed to heavy metals exude high levels of
phenolics (Winkel-Shirley 2002).
Tannin-rich plants such as tea, which are tolerant to
excess manganese levels, are protected by the direct chela-
tion of manganese. Direct chelation and binding to poly-
phenols was observed with methanol extracts of rhizome
polyphenols from Nympheae for Cr, Pb, and Hg (Lavid and
others 2001). According to Morgan and others (1997), this
general chelating ability of phenolic compounds is probably
related to the high nucleophilic character of the aromatic
rings rather than to specific chelating groups within the
molecule. Metal ions decompose lipid hydroperoxide
(LOOH) by the homolytic cleavage of the O—O bond and
yield lipid alkoxy radicals, which initiate free radical chain
oxidation. Phenolic antioxidants inhibit lipid peroxidation
by trapping the lipid alkoxy radical. This activity depends on
the structure of the molecules and on the number and position
of the hydroxyl groups in the molecules (Milic and others
1998). Arora and others (2000) showed that phenolics,
especially flavonoids, are able to alter peroxidation kinetics
by modifying the lipid packing order. They stabilize mem-
branes by decreasing membrane fluidity, hinder the diffusion
of free radicals, and restrict peroxidative reactions. Through
such interactions, flavonoids help maintain membrane
integrity by preventing the hydrophobic region of the bilayer
from being accessed by the deleterious molecules. On the
other hand, in vitro studies have shown that flavonoids can
directly scavenge ROS such as superoxide, hydrogen per-
oxide, hydroxyl radical, singlet oxygen, and peroxyl radical
(Sakihama and others 2000). Their antioxidant action is
mainly the result of their ability to donate electrons or
hydrogen atoms. Polyphenols possess the ideal chemical
structure for this activity and have been shown to be more
effective in vitro than vitamins E and C on a molar basis
(Rice-Evans and others 1997). The presence of 3-OH group
in their structure is the most significant determinant of
electron-donating activity. The hydrogen peroxide-depen-
dent oxidation of flavonols has been observed in situ in
epidermal strips of leaves of Vicia faba and Tradescantia
virginiana and in mesophyll cells of V. faba (Takahama
1998a, b).
Nitrogen-Containing Secondary Metabolites and Plant
Defence
Increasing evidence from a variety of sources indicates that
the basic nitrogen-containing compounds in higher plants
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include potent inhibitors of various toxic oxidative com-
pounds. Several alkaloids of various structural types have
been found to be potent inhibitors of singlet oxygen. Par-
ticularly effective are indole alkaloids such as strychnine
and brucine that have a basic nitrogen atom in a rigid, cage-
like structure. Such alkaloids appear to be strictly physical
quenchers and are not destroyed chemically by the process
of quenching (Larson 1988). Thus, in principle, they could
inactivate many molecules of singlet oxygen per molecule
of alkaloid. Some of the alkaloids may be synthesized from
polyamines such as putrescine. Polyamines (spermidine
and spermine) play a variety of physiological roles in plant
growth and development (Ali 2000). They are also potent
ROS scavengers and inhibitors of lipid peroxidation
(Drolet and others 1986).
Alkaloids of the quinolizidine type, for example, spar-
teine, have been reported to be stored in the epidermal cells
of four plants belonging to the genus Lipinus. Ali (2000)
suggested that the storage pattern of various alkaloids was
consistent with a phytochemical role for these substances
as antifeedant chemical defence compounds, but it would
also be consistent with an antioxidant role. It is unlikely
that these alkaloids are acting as UV light-filtering agents
because their absorption in the solar UV range would be
minimal. Alkaloids commonly accumulate in plant tissues
subjected to different types of stress. Nitrogenous protec-
tive compounds, other than alkaloids, found in plants are
substances like cynogenic glycosides and glucosinolates.
They are not themselves toxic, unlike some of the alka-
loids, but are readily broken down to give off poison, some
of which are volatile, when the plant is crushed. Hydrogen
cyanide is a well-known poisonous gas released by cyno-
genic glycosides. The presence of cynogenic glycosides
deters feeding by insects and other herbivores. Thus, these
compounds are believed to be responsible for pest resis-
tance. These compounds generally break down to produce
molecules responsible for the smell and taste of vegetables
such as radish and broccoli, as these compounds are prin-
cipally found in Brassicaceae. Glucosinolates break down
to release defensive substances. The hydrolytic products of
glucosinolates function in defence as herbivore toxins and
feeding repellents (Taiz and Zeiger 2006).
Brassinosteroids
Brassinosteroids are a relatively new group of plant hor-
mones involved in significant growth-promoting activities.
They are polyhydroxysteroids structurally similar to animal
and insect steroid hormones. More than 60 brassinosteroids
have been identified and characterized so far from various
plant organs, including pollens, seeds, and vegetative
shoots (Bajguz and Tretyn 2003; Wang and others 1993).
J Plant Growth Regul (2013) 32:216–232 225

Apart from occurring naturally, brassinosteroids can now
be synthesized in vitro. Brassinolide, the first isolated
brassinosteroid, can be synthesized from membrane sterol
(campesterol) through a series of reduction, hydroxylation,
epimerization, and oxidation steps. Recent evidence
suggests that the pathway is more of a metabolic grid than
two distinct branches. Many of the genes encoding bras-
sinosteroid biosynthetic enzymes have been cloned from
Arabidopsis thaliana, and mutants blocking several

Table 3 The occurrence of brassinosteroids in plants

Family/species Plant parts Brassinosteroids

Arecaceae
Phoenix dactylifera L. Pollen 24-epiCS
Gramineae
Lolium perenne L. Pollen 25-MeCS
Oryza sativa L. Shoot CS, DS, BL
Bran 6-DeoxoCS, 28-HomoTE, 28-HomoTY
Seeds CS, TE, 6-DeoxoCS
Phalaris canariensis L. Seeds CS, TE
Seeds CS, TY
Secale cereale L. Seeds CS, TY, TE, 6-DeoxoCS, 28-NorCS, SE
Triticum aestivum L. Grains CS, TY, TE, 6-DeoxoCS, 3-DT
Zea mays L.
Dent corn Pollen
Sweet corn Pollen CS, TY, TE
CS,28-NorCS, DS
Liliaceae
Erythronium japonicum Decne Pollen TY
Lilium elegans Thunb. Pollen BL, CS, TY, TE
Lilium longiflorum Thunb. Pollen BL, CS, TY
Anther 3-DT, TE-3-La, TE-3-My, TE-Glu
Tulipa gesneriana L. Pollen TY
Typhaceae
Typha latifolia G.F.W. Mey Pollen TY, TE
Betulaceae
Alnus glutinosa (L.) Gaertn. Pollen BL, CS
Cannabaceae
Cannabis sativa L. Seeds CS, TE
Caryophyllaceae
Gypsophilla perfoliata L. Seeds 24-epiBL
Lychnis viscaria L. Seeds 24-epiCS, 24-epiSE
Beta vulgaris L. Seeds CS, 24-epiCS
Fagaceae
Castanea crenata Sieb. et Zucc. Galls CS, BL, 6-DeoxoCS
Shoot CS
Leaves 6-DeoxoCS
Polygonaceae
Fagopyrum esculentum Moench Pollen BL, CS
Rheum rhabarbarum L. Panicles BL, CS, 24-epiCS
Apiaceae
Apium graveolens L. Seeds 2-DeoxyBL
Daucus carota ssp. sativus L. Seeds BL, CS, 24-epiCS

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226 J Plant Growth Regul (2013) 32:216–232

Table 3 continued
Family/species Plant parts Brassinosteroids

Brassicaceae
Arabidopsis thaliana (L.) Heynh. Shoot CS
Ecotype Columbia (wild type) 6-DeoxoCS, TY, 6-DeoxoTY, BL, 28-NorCS,
28-NorTY, TE, 6-DeoxoCT, 6-DeoxoTE,
3-Dehydro-6-DeoxoTE
Seeds BL
Ecotype Columbia (Wild type) 24-epiBL, CS, 6-DeoxoCS, TY, 6-DeoxoTY, 6-
Seeds (ecotype 24) DeoxoTY
Root Callus 24-epiBL, CS
BL, 3-epiBL
Brassica campestris Var. Seeds BL, 28-NorBL, CS, 28-
pekinensis L. NorCS, 28-HomoCS
Brassica napus L. Pollen BL
Raphanus sativus L. Seeds BL, CS, TE, 28-HomoTE
Fabaceae
Cassia tora L. Seeds BL, CS, TY, TE, 28-NorCS
Dolichos lablab L. Seeds DL, DS, 28-HomoDS, 28-HomoDL, BL, CS,
6-DeoxoCS, 6-DeoxoDS
Robinia pseudo-acacia L. Pollen CS, TY, 6-DeoxoCS
Vicia faba L. Seeds BL,24-epiBL,CS,28-NorCS
Pollen BL,CS,28-NorCS, DS
Psophocarpus tetragonolobus (Stickm.) DC. Seeds BL, CS, 6-DeoxoCS, 6-DeoxoDS
Ornithopus sativus Brot. Seeds CS, 24-epiCS
Shoot CS, 6-DeoxoCS, 24-epiCS, 6-Deoxo-24-epiCS,
6-Deoxo-28-NorCS
Phaseolus vulgaris L. Seeds BL, CS, 2-epiCS, 3-epiCS, 2,3-DiepiCS, 3,24-
DiepiCS, TY, TE, 6-DeoxoCS, 3-epi-6-DeoxoCS,
1b-OH-CS, 3-epi-1a-OH-CS, DL, DS,
6-DeoxoDS, 6-Deoxo-28-HomoDS, 25-MeDS,
2-epi-25-MeDS, 2,3-Diepi-25-Me
DS, 2-Deoxy-25-MeDS, 2-epi-2-Deoxy-25-MeDS,
2-Deoxy-25-MeDS, 3-epi-2-Deoxy-25-MeDS,
6-Deoxo-25-MeDS, 25-MeDS-Glu, 2-epi-25-
MeDS-Glu
Pisum sativum L. Seeds BL, CS, TY, 6-DeoxoCS, 2-DeoxyBL
Shoot BL, CS, 6-DeoxoCS, TY, 6-DeoxoCT, 6-DeoxoTE,
3-Dehydro-6-DeoxoTE, 6-DeoxoTY
Myrtaceae
Eucalyptus calophylla R.Br. Pollen BL
Eucalyptus marginata Sn. Pollen DS
Rosaceae
Eriobotrya japonica (Thunb.) Lindl. Flower buds CS
Rutaceae
Citrus unshiu Marcov. Pollen BL, CS, TY, TE
Citrus sinensis Osbeck Pollen BL, CS
Theaceae
Thea sinensis Leaves 28-NorCS, 28-HomoCS, BL, CS, TY, TE
Apocynaceae
Catharanthus roseus G. Don. Cultured cell BL, CS, 6-DeoxoTY, 6-DeoxoTE, 6-DeoxoCS, CT,
6-DeoxoCT, 6-epi-6-DeoxoCT, 3-DT, TY, TE

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J Plant Growth Regul (2013) 32:216–232 227

Table 3 continued
Family/species Plant parts Brassinosteroids

Asteraceae
Zinnia elegans L. Cultured cell CS, TY, 6-DeoxoCS, 6-DeoxoTY, 6-DeoxoTE
Helianthus annuus L. Pollen BL, CS, 28-NorCS, BL
Solidago altissima L. Shoot BL
Boraginaceae
Echium plantagineum L. Pollen BL
Convolvulaceae
Pharbitis purpurea Voigt Seeds CS, 28-NorCS
Cucurbitaceae
Cucurbita moschata Duch. Seeds BL
Lamiaceae
Perilla frutescens (L.) Britt. Seeds CS
Solanaceae
Nicotiana tabacum L. Cultured cell CS
Lycopersicon esculentum Mill. Shoot CS, 6-DeoxoCS, 28-NorCS
Root 6-Deoxo-28-NorCT, 6-Deoxo-28-NorTY, 6-Deoxo-
28-NorCY
Cupressaceae
Cupressus arizonica Greene Pollen 6-DeoxoTY, 6-DeoxoCS, 3-Dehydro-6-DeoxoTE,
CS, TY, TE, BL, 3-DT, 28-HomoCS
Ginkgoaceae
Ginkgo biloba L. Seeds TE
Pinaceae
Piceae sitchensis Trantv.ex Mey Shoot CS, TY
Pinus silvestris L. Cambial region BL, CS
Pinus thunbergii Parl. Pollen TY
Taxodiaceae
Cryptomeria japonica D. Don. Pollen TY
Anther DL, 3-DT, 28-HomoBL,
28-HomoDL, 23-DehydroBL (cryptolide), 2-epi-23-
DehydroBL, 3-epi-23-DehydroBL, 2,3-Diepi-23-
DehydroBL
Equisetaceae
Equisetum arvense L. Whole plant CS, DS, 28-NorBL, 28-NorCS
Hydrodictyaceae
Hydrodictyon reticulatum (L.) Lager. Whole plant 24-epiCS, 28-HomoCS
Marchantiaceae
Marchantia polymorpha L. Cultured cell TE, 3-DT, TY
CS castasterone, TY typhasterol, DL dolicholide, BL brassinolide, TE teasterone 3-dehydroteasterone, SE secasterone, DS dolichosterone

biosynthetic steps have been identified in Arabidopsis, pea,
and tomato (Shimada and others 2001).
Distribution
There are about 58 plant species, including 49 angio-
sperms (12 monocotyledons and 37 dicotyledons), 6 gym-
nosperms, 1 pteridophyte (Equisetum arvense), 1 bryophyte
(Marchantia polymorpha), and 1 chlorophyte (Hydrodictyon
reticulatum), from which brassinosteroids have been char-
acterized. As brassinosteroids are found to be present in
almost all the plants tested so far, they are probably ubiqui-
tous in the plant kingdom. Some of the plants and their parts
in which brassinosteroids are located, are given in Table 3
(Bajguz and Tretyn 2003). Brassinosteroids are normally
present in extremely low concentrations and their levels vary

123
228
Fig. 4 Brassinosteroids and
stress tolerance
among different plant tissues. Young growing tissues contain
higher levels of brassinosteroids than mature tissues. Pollen
and immature seeds are the richest sources of brassinoster-
oids, with a concentration range of 1–100 ng/g fresh weight,
whereas shoots and leaves usually have lower amounts, that
is, 0.01–0.1 ng/g fresh weight (Takatsuto 1994).
Physiological Functions
Brassinosteroids are a class of plant steroid hormones
whose general effect is the promotion of cell elongation,
cell division, differentiation, disease resistance, stress tol-
erance, and senescence throughout the plant life cycle.
Thus, brassinosteroids possess significant growth-control-
ling activity. Other physiological responses to brassinos-
teroids include reproductive and vascular development,
membrane polarization, and proton pumping. They also
influence various other developmental processes like seed
germination, rhizogenesis, and flowering. Due to multiple
effects, brassinosteroids are considered plants hormones
with pleiotropic effects (Fig. 4).
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J Plant Growth Regul (2013) 32:216–232
Brassinolide, castasterone, and 24-epibrassinolide are
among the naturally occurring brassinosteroids that are
considered to be the most important ones due to their wide
distribution as well as their potent biological activity. They
can induce resistance to environmental stresses such as
drought, extreme temperature, heavy metals, herbicide
injury, and salinity. Exogenous application of brassinos-
teroids modifies the activity of antioxidant enzymes such as
SOD, CAT, glutathione peroxidase, and APX, and it affects
nonenzymatic antioxidants such as ascorbic acid, tocoph-
erol, carotenoids, and glutathione in plants under different
stress conditions. Brassinosteroids may influence a range of
diverse processes of growth and development in plants
when applied exogenously (Cao and others 2005), regulate
the expression of numerous genes, and affect the activity of
complex metabolic pathways (Kartal and others 2009). An
increase in crop yield and quality has been observed when
plants were treated with exogenous brassinosteroids at
appropriate stages of their development (Singh and Shono
2005). Plants that suffer from oxidative stress generate a
high overabundance of reducing equivalents. Despite the
J Plant Growth Regul (2013) 32:216–232
fact that massive amounts of NADPH ? H? are oxidized
by photorespiration and the xanthophyll cycle, under such
stressful conditions the corresponding strong reduction
power seems to enhance the synthesis of highly reduced
compounds like isoprenoids, phenols, or alkaloids. Con-
sequently, the synthesis and accumulation of highly
reduced secondary plant products prevents massive gen-
eration of oxygen radicals and the corresponding damage
by photoinhibition. Because of their significant ROS-
scavenging capacity, it is proposed that along with bras-
sinosteroids, the phenylpropanoid-derived phenols, that is,
flavonoids, tannins, and hydroxycinnamate esters, which
are produced in the course of various stress situations,
represent important radical scavengers (Selmar 2008).
Modulation of Stress Responses
Increased tolerance of brassinosteroid-treated plants to
environmental stresses may be attributed to the fact that
brassinosteroids induce changes in membrane stability and
osmoregulation, leading to improved stress tolerance
(Wang and others 1993). During chilling at 1-5 C, the
application of 24-epibrassinolide to rice plants reduced
electrolyte leakage and malondialdehyde content and
enhanced the levels of proline (Clouse and Sasse 1998).
Treatment of tomato plants with 24-epibrassinolide
enhanced tolerance to high temperature and induced the
expression of mitochondrial small heat shock proteins,
which possibly are responsible for improved thermotoler-
ance (Singh and Shono 2005). 24-Epibrassinolide protects
the leaf cell ultrastructure in rice under salt stress and
prevents nuclei and protoplast degradation (Wang and
others 1993). Application of brassinosteroids has been
reported to improve drought stress tolerance in sugar beet
and wheat (Sairam 1994). These phytohormones enhance
resistance to salt, cold, fungal infection, and herbicide
injury in plants (Clouse and Sasse 1998). Removal of the
inhibitory effect of salt stress on pigment levels with
brassinosteroid treatment has been reported; this could be
one of the reasons for growth stimulation under saline
conditions (Anuradha and Rao 2003). Houimli and others
(2010) also reported a positive effect of the external
application of brassinosteroid in pepper under salinity
stress. Proline content in brassinosteroid-treated mung bean
hypocotyl segments was remarkably enhanced under stress
conditions (Zhao and Chen 2003). The above-mentioned
studies reported that 24-epibrassinolide enhances the
resistance of Pseudomonas syringae pv. tabaci (Pst),
Oidium sp., and Oxyza sp. to viral, bacterial, and fungal
pathogens, respectively. Brassinosteroids lead to increased
synthesis of ethylene and hence activate the protective
substances of phenolic and terpenoid nature (Korableva
and others 2002). On analysing the inhibitor brassinazole
229
2001 for biosynthesis of brassinosteroids and the mea-
surement of brassinosteroids in TMV-infected tobacco
leaves, it was observed that steroid hormone-mediated
disease resistance (BDR) plays a part in the defence
response in tobacco. In previous studies, brassinolide
treatment enhanced resistance against various pathogens in
both tobacco and rice, suggesting that brassinolide func-
tions as one of the common signalling molecules in the
innate immunity system of higher plants (Nakashita and
others 2003). It has been suggested that BAK1 may act as a
coreceptor for several receptor-like kinases (RLKs),
thereby controlling various processes such as development,
cell death, and perception of pathogen-associated molecu-
lar pattern (PAMP) signalling in a stimulus-dependent
manner (Kemmerling and Nürnberger 2008). Furthermore,
SERK3/BAK1 (somatic embryogenesis receptor kinase
3/BRI1-associated receptor kinase 1) and SERK1 both
interact with BRIl to facilitate brassinosteroid signalling
(Karlova and others 2000). It is also reported that the
expression of pathogenesis-related genes is greatly reduced
in the case of brassinosteroid-deficient mutants (Szekeres
and others 1996).
Mode of Action
Brassinosteroids are present at low levels throughout the
plant kingdom and regulate the growth and developmental
processes in young growing tissues. Studies suggest that
brassinosteroids affect the mechanical properties of the cell
wall during cell elongation. The phenotypes of brassinos-
teroid-deficient and -insensitive mutants confirm that
brassinosteroids are essential for cell elongation and also
suggest a role in vascular differentiation, senescence, fer-
tility, leaf morphology and light–dark regulation of
development (Clouse 2001).
The signal transduction pathway involving brassinos-
teroids has been studied in Arabidopsis using a brassinos-
teroid-insensitive mutant. This mutant, named bril, showed
severe pleiotropic effects upon development, inducing
dwarfism, de-etiolation, male sterility, and altered leaf
morphology (Clouse and others 1996). The Arabidopsis
genome contains brassinosteroid receptors that have spe-
cific functions in cell growth and vascular differentiation.
Recently, a few brassinosteroid receptors have been iden-
tified from A. thaliana, some of which are leucine-rich
proteins. The brassinosteroid receptor (BRI1) has a kinase
domain and is located on the plasma membrane (unlike in
the case of animal systems, where receptors for steroid
hormones are intracellular); it functions at the cell surface
and transduces extracellular signals. A hypothetical scheme
for brassinosteroid signal transduction proposes that the
binding of brassinosteroid molecules to the receptor causes
the activation of a kinase domain, and, subsequently,
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230
phosphorylation of additional kinases occurs (Clouse and
Sasse 1998). Higher expression of BRI1 is reported in
shoots, roots, hypocotyls of seedlings, and meristem. It is
also expressed at comparatively lower levels in the later
developmental stages. Previously reported studies con-
firmed BRI1, which is a leucine-rich repeat receptor, as
Ser/Thr kinase and suggested that it plays a role in bras-
sinosteroid signalling through Ser/Thr phosphorylation
(Friedrichsen and others 2000). The protein BRI1-associ-
ated receptor kinase (BAK1) has been reported to interact
with BRI1 and participate in brassinosteroid signalling,
sharing structural organization with BRI1. This protein was
also found to be localized on the plasma membrane, as is
BRI1. Direct physical interaction between both proteins
has been confirmed in yeast cells and Arabidopsis (Li and
others 2002; Nam and Li 2002). Interestingly, besides
brassinosteroids, tomato BRI1 (tBRI1) has been found to
act as the receptor for a peptide hormone called systemin
that is known to mediate the response to wounding by
insect pests (Scheer and others 2003). Genetic studies have
also led to the identification of brassinosteroid insensitive
kinase 2 (BIN2), which is similar to SHAGGY/GSK3
kinases (glycogen synthase kinase 3) in animal systems, as
a negative regulator, and two nuclear proteins, brassinazole
resistant 1 (BZR1) and bri1-EMS suppressor (BES1), as
positive regulators in the brassinosteroid signalling path-
way. The accumulation of BZR1 and BES1 is reported to
be increased by brassinosteroid treatment (Choe and others
2002; Yin and others 2002). Although a number of
important regulatory proteins involved in the brassinos-
teroid signalling pathway have been isolated, links con-
necting these proteins are yet to be identified (Zhou and
others 2004). Further research may be able to bridge the
gaps in our understanding. These studies raise the possi-
bility of conserved interaction between brassinosteroid
signalling, defence response, and peptide signalling
pathways.
Conclusions
Plant secondary metabolites present an interesting class of
biological molecules with extremely diverse and important
functions. A decrease in arable land due to anthropogenic
causes and the necessity to grow more food on the limited
available land have brought into focus the role of second-
ary metabolites as plant protectors and tools for enhancing
crop productivity. The importance of secondary metabo-
lites is a consequence of their diversity of structure and
function, as they are of critical importance for the survival
of plants under stress. Advancements in cell culture tech-
niques have allowed in vitro production of bioactive sec-
ondary metabolites, in controlled environments and
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J Plant Growth Regul (2013) 32:216–232
independent of climate and soil conditions, thus providing
a continuous and reliable source of natural products. The
improved use of biotechnological tools and an emerging
picture of the structure and regulation of pathways for
secondary metabolite production provide the basis for the
production of commercially acceptable levels of secondary
metabolites. Along with secondary metabolites, brassinos-
teroids present a class of potent plant growth regulators of
steroidal nature with immense agricultural potential.
Brassinosteroids have significant growth-promoting activ-
ity and are of critical value when plants are exposed to
stress. Structurally modified brassinosteroids with greater
stability under field conditions have been synthesized and
tested. An important role of brassinosteroids in realizing
better crop yields in the 21st century is inevitable.
However, strategies are needed to develop an informa-
tion database that would help researchers understand how
synthesis of bioactive secondary metabolites could be
enhanced at the cellular and molecular levels. The intro-
duction of recombinant DNA technology has led to the
production of transgenic cells/tissues that streamline the
regulation and production of secondary metabolites. This is
a significant step towards making cell cultures more ame-
nable to commercial production of secondary metabolites.
It will also help in unravelling the molecular mode of
action of these compounds. Use of secondary metabolites
and brassinosteroids for plant protection under stress is a
promising field that holds promise for the food and nutri-
tional security of the world.
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