Crop Protection 29 (2010) 913e920

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Crop Protection
journal homepage: www.elsevier.com/locate/cropro

Review

Mechanism of action of insecticidal secondary metabolites of plant origin

Rameshwar Singh Rattan*
Entomology and Pesticide Residue Analysis Laboratory, Hill Area Tea Sciences (HATS) Division, Institute of Himalayan Bioresource Technology (CSIR), Post Box-6, Palampur 176061,
Himachal Pradesh, India

a r t i c l e i n f o a b s t r a c t

Article history: Insect pest management is facing the economic and ecological challenge worldwide due to the human
Received 20 November 2009 and environmental hazards caused by majority of the synthetic pesticide chemicals. Identification of
Received in revised form novel effective insecticidal compounds is essential to combat increasing resistance rates. Botanicals
12 May 2010
containing active insecticidal phytochemicals appear to be promising to address some of these problems.
Accepted 13 May 2010
Therefore, there is a continuous need to explore new active molecules with different mechanisms of
action. Secondary metabolites present in plants apparently function as defense (toxic), which inhibits
Keywords:
reproduction and other processes. The phytochemical biomolecules could be used for maximizing the
Chemical ecology
Insecticide
effectiveness and specificity in future insecticide design with specific or multiple target sites, while
Botanicals ensuring the economic and ecological sustainability. In this article, the current state of knowledge on
Plant allelochemicals phytochemical sources and insecticidal activity, their mechanism of action in insects, resistance, and
Toxicity promising advances made in phytochemical research are reviewed.
Plant protection Ó 2010 Elsevier Ltd. All rights reserved.
Insect pest management
Mechanism of action
Acetylcholine
Octopamine
GABA

1. Introduction management approaches. In addition, they are playing a vital role in

Insecticides are the cornerstones upon which the pest
management practices are based, and are likely to remain so as long
as effective and inexpensive chemicals are available (Haynes, 1988).
The pest management in agriculture is facing challenge in devel-
opment of suitable agents to kill insect pests while ensuring the
economic and ecological sustainability as majority of the pesticide
chemicals are known to cause human and environmental hazards.
In the recent past, a variety of new insect control agents have been
developed, or are being developed, which may fit a variety of insect
pest management needs, and many of these have novel mechanism
of action. The growing demand for natural products has intensified
in the past decades as they are extensively used as biologically
active compounds and, are being considered an important alter-
native strategy for the sustainable insect pest management in
agriculture, as they are biodegradable and potentially suitable for
use in integrated management programs. Pesticides of plant origin
are gaining increased attention and interest among those con-
cerned with environment friendly, safe and integrated crop
* Tel.: þ91 1894 33339(x)327; fax: þ91 1894 230433/230428.
E-mail address: rsrattan@scientist.com
organic food production globally.
Insecticides exert a broad range of effects on insects and other
arthropods viz. neuroexcitation resulting in hyperactivity, tremor
and rigid paralysis due to energy depletion and neuromuscular
fatigue, while neuroinhibition results in immobility and paralysis
because of possible oxygen deprivation and/or reduced respiratory
capacity that ultimately leads to mortality (Scharf et al., 2003). It is
important to investigate behavioral patterns in insects to elucidate
the mode of action of novel and conventional insecticides, and their
response in the environment by minimizing their contact with the
toxic material (von Keyserlingk et al., 1985). Multiple biological
signals and mechanisms regulate the efficacy of the synaptic
transmission, thus providing rich combinational possibilities for
modifying neural communication. The bio-regulation and mecha-
nisms has been studied by various workers, especially for synthetic
pesticides, which provide a high degree of precision in modification
of neural pathways (Evans and Robb, 1993; Roeder et al., 1995; Von
Nickisch-Rosenegk et al., 1996; Grohmann et al., 2003; Balfanz
et al., 2005; Maqueira et al., 2005; Ozoe and Huang, 2008).
Plants have evolved a variety of defense mechanisms to reduce
insect attack, both constitutive and inducible, while insects have
evolved strategies to overcome these plant defenses. Chemical
ecology is the study of how particular chemicals (allelochemicals)

0261-2194/$ e see front matter Ó 2010 Elsevier Ltd. All rights reserved.
doi:10.1016/j.cropro.2010.05.008
914 R.S. Rattan / Crop Protection 29 (2010) 913e920
are involved in interactions of organisms with each other and with
their surroundings. Such allelochemicals function as chemical
defense compounds and influence molecular targets in herbivores
or microbes. These are useful for plants against most vertebrates,
since the elements of neuronal signaling pathways are quite similar
throughout the animal kingdom. The mixture of secondary
metabolites may be deterrent to insects and herbivores for longer
period than single compound and, different physical properties
may allow more deployment or longer persistence of defenses. The
mode of action and site of effect for insecticidal activities have been
studied by various authors (Lewis et al., 1993; Roeder, 1994; Vanden
Broeck et al., 1995; Zafra-Polo et al., 1996; Enan, 1998, 2001;
Kostyukovsky et al., 2002; Priestley et al., 2003). Mostly the work
has been carried on studying the effects of extracts and essential
oils, their lethal doses and time to achieve lethal effects, but mode
of action are in general not fully elucidated. Since, large numbers of
chemical defenses are present in the nature; very little is known
about their mode of action at molecular level. In this review,
phytochemical sources of insecticidal activity, their mode of action,
molecular targets and their impact on development of resistance
along with future developments are discussed.
2. Mechanism of action: phytochemicals
There is no absolute way to find a new mode of action, and
serendipity is usually more important than biorational approaches;
it is facilitated by enzyme, ion-channel and membrane target assays
as supplements to determination of insecticidal activity (Casida and
Quistad, 1998). The most successful plant species typically
synthesize a wide array of moderately toxic defense compounds or
a small number of highly toxic substances. The insect herbivores
feed on a wide range of plant species, potentially encountering
toxic substances with relatively non-specific effects on a wide range
of molecular targets. These targets range from proteins (enzymes,
receptors, ion-channels, structural proteins), nucleic acids, bio-
membranes to secondary metabolites with specific or unspecific
interactions and other cellular components (Harborne, 1993).
Analogues of secondary metabolites interfere with certain vital
components of the cellular signaling system, or interfere with vital
enzymes, signaling in the nervous system (such as neurotrans-
mitter synthesis, storage, release, binding, and re-uptake, receptor
activation and function, enzymes involved in signal transduction)
and block metabolic pathways (Wink, 2000). The knowledge of
chemical properties of such novel compounds is necessary to
determine the safety and economics of their use in agriculture.
However, due to observed antifeedant and growth regulation
effects (Nathan et al., 2004), it is apparent that botanical insecti-
cides affect insect physiology in many different ways and at various
receptor sites (Table 1). In lepidopteran larvae, terpenes (drimane
sesquiterpines) block the stimulatory effects of glucose and inositol
on chemosensory receptor cells located on the mouthparts, and
they could also act on receptors in other ways (Gershenzon and
Dudareva, 2007). Essential oils and their constituents affect
biochemical processes, which specifically disrupt the endocrino-
logic balance of insects. They may be neurotoxic or may act as insect
growth regulators, disrupting the normal process of morphogen-
esis (Reynolds, 1987; Balandrin and Klocke, 1988). The neurotox-
icity of several monoterpenoids (D-limonene, myrcene, terpineol,
linalool and pulegone), which have been identified as important
components of essential oils, were evaluated against the house fly
as well as on the German cockroach (Coats et al., 1991). Singh and
Aggarwal (1988) found Himachallol and b-Himachalene toxic to
pulse beetle. Toxicity from essential oils or their constituent in
insects and other arthropods points to a neurotoxic mode of action;
most prominent symptoms are hyperactivity followed by hyper-
excitation leading to rapid knock down and immobilization (Enan,
2001).
Allelochemical defense system in insects include P450, Gluta-
thione-S-transferases and esterases, which are typically concen-
trated in insect’s midgut, allowing rapid elimination of ingested
toxic substances, and/or the fat bodies, facilitating the detoxifica-
tion of contaminants penetrating cuticular or tracheal structure
(Schuler, 1996). Avermectin and milbemycin insecticides act at
gultamate gated chloride channels acting as agonists of chloride
channels, which elicit increased chloride current flow into neurons
(Bloomquist, 2001). Pyrethrins give knock down action, which in
many cases is irreversible and could lead to paralysis (Casida, 1973).
Knock down may be caused by hyper-excitation of the nerve and
nerve blockage is responsible for paralysis. Generally, pyrethrins
and pyrethroids effects sodium channel by activation at lower
threshold; which result in prolonged flow of sodium currents into
neurons (Evans, 1984; Sonderlund, 1995). The mode of action of
Sabadila (Schoenocanlon offiicinale) is similar to that of pyrethrins,
as it affects the voltage dependent sodium channels of nerve axon
(Ohta, 1973; Levi et al., 1980). Nicotine poisoning symptoms follow
the general sequence of excitation, convulsions, paralysis, death
and mimics as a part of the action of acetylcholine by interacting
with the nicotine acetylcholine receptor as receptor fails to
distinguish between nicotine and acetylcholine (Richards and
Cutkomp, 1945). Veratridine causes an increase in the duration of
the action potential, repetitive firing, and a depolarization of the
nerve membrane potential (Bloomquist, 1996). In insects, the result
of this nerve poisoning can be immediate death or several days of
paralysis before death. Rotenone (from Deris eliptica and Loncho-
carpus species) disrupts energy metabolism in cell mitochondria,
either by inhibiting the electron transport system or by uncoupling
the transport system from ATP production (Fukami, 1961;
Yamamoto and Kurokawa, 1970; Storey et al., 1981). It interferes
with electron transport system between NADH dehydrogenase and
coenzyme Q at complex1 as piericidin-A, causing a severe reduc-
tion in oxygen consumption. In insects, mouthparts become para-
lyzed so that they stop feeding and die of starvation. Rotenone gets
converted to metabolites that are highly toxic to insects but it
detoxifies in mammals (Ware, 1988). The poisoning symptom
appears in sequence as inactivity, locomotive instability, knock
down, paralysis and slow death. Recently, azadirachtin was cate-
gorized as an anti-mitotic (G2/M phase of cell division) insecticide
(Salehzadeh et al., 2003). Studies using radio-labeled azadirachtin
have indicated its specific localizations to corpus cardiacum
(Rembold et al., 1989). On the other hand, Kumar et al. (2007)
showed that few scattered amino acid changes caused significant
steric hindrance in the binding pocket for b-actin, causing a reverse
orientation of azadirachtin. The single point mutation in the Rdl
GABA receptor subunit of Ala302 to Ser confers resistance to diel-
drin in Drosophila, which is also resistant to a-Thujone to a lesser
degree (ffrench Constant and Roush, 1991; ffrench Constant et al.,
1993; Hold et al., 2000). The major targets of insect neuro
systems are discussed below:
2.1. Cholinergic system
Acetylcholinesterase (AChE) plays role in cholinergic synapses
that is essential for insects and higher animals (Fournier and
Mutero, 1994). Inhibition of AChE causes accumulation of acetyl-
choline at the synapses, so that the post-synaptic membrane is in
a state of permanent stimulation, which results in ataxia i.e. general
lack of co-ordination in the neuromuscular system, and eventual
death (Singh and Singh, 2000; Aygun et al., 2002). It has been
reported that the essential oils act by a reversible competitive
inhibition of acetylcholinesterase (AChE) enzymes isolated from
 R.S. Rattan / Crop Protection 29 (2010) 913e920
Table 1
Mechanism of action of insecticides of plant origin.
S. System Mechanism of action Compound
No
1 Cholinergic Inhibition of acetylecholinestrase (AChE) Essential
system oils
Cholinergic acetylcholine nicotinic receptor Nicotine
agonist/antagonist
2 GABA system GABA-gated chloride channel Thymol,
Silphinenes
3 Mitochondrial Sodium and potassium ion exchange Pyrethrin
system disruption
Inhibitor of cellular respiration Rotenone
(mitochondrial complex I electron transport
inhibitor or METI)
Affect calcium channels Ryanodine
Affect nerve cell membrane action Sabadilla
4 Octopaminergic Octopaminergic receptors, Essential
system oils
Block octopamine receptors by working Thymol
through tyramine receptors cascade
Miscellaneous Hormonal balance disruption Azadirachtin Azadiractina indica
electric eel, heads of house fly, cockroaches, horse serum and
bovine erythrocytes (Grundy and Still, 1985; Ryan and Byrne, 1988;
Miyazawa et al., 1997; Keane and Ryan, 1999; Mills et al., 2004).
Recently, it was found that azadirachtin (a tetratepenoid) signifi-
cantly inhibits the activity of AChE in Nilaparvata lugens (Nathan
et al., 2008). AChE-EC 3.1.1.7 is a key enzyme responsible for
terminating the nerve impulse by catalyzing the hydrolysis of
neurotransmitters in the nervous system (Wang et al., 2004). AChE
have been observed to be organophosphorus and carbamate
resistant, and is well-known that the alteration in AChE is one of
main resistance mechanism in insect pests (Hama and Hosoda,
1983; Hemingway et al., 1986a,b; Kranthi et al., 2001; Wang et al.,
2004). Several essential oils from aromatic plants, monoterpenes
and natural products have been shown as inhibitors of AChE against
different insect species (Grundy and Still, 1985; Ryan and Byrne,
1988; Miyazawa et al., 1997; Kostyukovsky et al., 2002; Shaaya
and Rafaeli, 2007). Essential oil of Zingiber officinale was found to
alter the behaviour and memory in cholinergic system (Felipe et al.,
2008), while linalool was identified as an inhibitor of acetylcho-
linesterase (Ryan and Byrne, 1988). The alkaloids viz. berberine,
palmatine and sanguinarine, are toxic to insects and vertebrates,
which substantially affected acetylcholinesterase, butyr-
ylcholinesterase, choline acetyl-transferase, alpha 1- and alpha 2-
adrenergic, nicotinergic, muscarinergic and serotonin-2 receptors.
Also mulliken charge on the nitro-oxygen atom of 2-nitro-
iminoimidazolidines blocks neuronal activity in Periplaneta ameri-
cana, fluorine atom of 3-fluoropropyl-2-nitroiminoimidazolidine
acts as a hydrogen bond acceptor in its interaction with nAChRs,
and a molecule in which two imidacloprid molecules are tethered
with a hexamethylene linker is reported to be a nAChR antagonist
with high insecticidal activity (Kagabu, 2008). Sanguinarine
appears to be the most effective inhibitor of choline acetyl-trans-
ferase (IC50 284 nM) (Schmeller et al., 1997). Furthermore, alkaloids
of Haloxylon salicornicum and Stemona japonica are known be
strong agonists at nicotinic acetylcholine receptors (El-Shazly et al.,
2005; Tang et al., 2008). Berberine and sanguinarine intercalate
DNA; inhibit DNA synthesis and reverse transcriptase. In addition,
sanguinarine (but not berberine) affects membrane permeability
and berberine protein biosynthesis (Schmeller et al., 1997). The
insect’s acetylcholinesterase (AChE) gene encoding Ace was cloned
from Drosophila (Hall and Spierer, 1986; Hall and Malcolm, 1991).
The Ace gene replacements linked to AChE insensitivity in insects e.
915
Plant source References
Azadirachtina indica, Mentha spp., Grundy and Still, 1985; Ryan and Byrne, 1988;
Lavendula spp. Miyazawa et al., 1997; Keane and Ryan, 1999; Felipe
et al., 2008
Nicotiana spp., Delphinium spp., Richards and Cutkomp, 1945; Kukel and Jennings,
Haloxylon salicornicum, Stemona 1994
japonicum
Thymus vulgaris Priestley et al., 2003; Ratra and Casida, 2001;
Bloomquist et al., 2008
Crysanthemum cinerariaefolium Casida, 1973
Lonchocarpus spp. Yamamoto and Kurokawa, 1970; Ware, 1988; Storey
et al., 1981; Khambay et al., 2003
Ryania spp. Copping and Menn, 2000
Schoenocaulon officinale Bloomquist, 1996
Cedrus spp., Pinus spp., Citronella Enan, 2005a; Nathanson et al., 1993; Kostyukovsky
spp., Eucalyptus spp. et al., 2002
Thymus vulgaris Enan, 2005a; Enan, 2005b
g. the Colorado potato beetle Leptinotarsa decemlineata (Zhu and
Clark, 1997).
2.2. GABA system
Ligand-gated chloride channels are proven target sites for
insecticides acting as antagonists by stabilizing non-conducting
conformations of the chloride channel. Blockage of the GABA-gated
chloride channel reduces neuronal inhibition, which leads to
hyper-excitation of the central nervous system, convulsions, and
death (Bloomquist, 2003). GABA and related aminobutyric acids are
known to stimulate feeding and evoke taste cell responses among
herbivorous insects and these allelochemicals antagonize GABA
phagostimulants, thereby, inducing feeding deterrence (Mitchell,
1987; Mullin et al., 1994). Pyrethrin (cyclopropyl monoterpene
esters) from Chrysanthemum cinerarifolium disrupts the insect
nervous system by acting on the voltage sensitive sodium channel
of the nerve membrane (Sonderlund, 1995). Another possible target
suggested for essential oils is the interference with GABA-gated
chloride channels in insects (Priestley et al., 2003). Thujone has
been classified as a neurotoxic insecticide, which acts on GABAA
receptors (Hold et al., 2000; Ratra and Casida, 2001). Thujone is
a competitive inhibitor of [3H]EBOB binding (i.e. of the non-
competitive blocker site of the GABA-gated chloride channel) and is
a reversible modulator of the GABAA receptor (Hold et al., 2000). It
was suggested that thymol potentiates GABAA receptors through an
unidentified binding site (Priestley et al., 2003). The silphinenes (a
plant-derived natural compounds) antagonize the action of GABA
on insect neurons (Bloomquist, 2003; Bloomquist et al., 2008).
2.3. Mitochondrial system
Insecticidal poisoning also occurs via affecting the activity of
protein phosphorylation (H-ATP: proton pump) or some enzyme
system (such as ATPase) (Cheng and Fu, 1989). Rotenone is a well-
known potent mitochondrial poison, which acts by inhibiting the
mitochondrial activity (Haley, 1978). Bullatacin (compound from
plants of the Annonaceae) shows strong inhibition of mitochondrial
electron transport with a specific action at complex I
(Ahammadsahib et al., 1993). Mode of action studies revealed that
dunnione exhibits both insecticidal and fungicidal activity by
inhibiting mitochondrial Complex III (Khambay et al., 2003). They
916 R.S. Rattan / Crop Protection 29 (2010) 913e920
are found throughout the plant kingdom and exhibit toxicity by
binding to proteins (Schultz, 1989).
2.4. Octopamenergic system
Octopamine (OA) is a target for essential oils activity in insects.
OA is a naturally occurring multifunctional biogenic amine, which
plays key roles as a neurotransmitter, neuromodulator and neuro-
hormone in invertebrate system, with a physiological function as
that of nornephrine in vertebrates (Evans, 1981). The acute and sub-
lethal behavioral effects of essential oil compounds on insects and
other vertebrates are consistent with an octopamenergic target site
in insects, which acts by blocking octopamine receptors (Evans,
1984; Enan, 2001, 2005b). The role of octopamine in invertebrate
system and metabolism, pharmacology, physiology, effect on
neuronal system, receptors and their mode of action have been
reviewed (Evans and Robb, 1993; Roeder, 1999). Physiological
functions of OA appears to be mediated by pharmacologically
distinct class of octopamine receptors viz. OCTOPAMINE receptors
(myogenic rhythm via Caþ concentration), OCTOPAMINE2A and
OCTOPAMINE2B, OCTOPAMINE3 (activation of adenylate cyclase
activity), which are coupled to different second messenger system
belonging to the family of G-protein coupled receptors (G-PCRs)
(Ryan and Byrne, 1988; Evans and Robb, 1993; Howell and Evans,
1998; Enan, 2001; Kostyukovsky et al., 2002). Pharmacological,
biochemical and physiological studies have supported the presence
of octopamine receptor sub-types in different species viz. American
cockroaches (Enan, 1998, 2005b), Bombyx mori (Aoyama et al.,
2001), Drosophilla melanogaster (Evans et al., 1995) and fruit flies
(Enan, 2005b). Enan (2001) also showed that low doses of eugenol
(106 M) caused a significant increase in adenosine cyclic mono-
phosphate (cAMP) in the nervous system of P. americana, a similar
effect to that of 106 M Octopamine. It was subsequently shown that
eugenol (2.5  105 M) mimicked octopamine (105 M) in increasing
intracellular calcium levels in cloned cells from the brain of P.
americana and D. melanogaster, and this was also found to be
mediated via octopamine receptors (Enan, 2005b). Pharmacolog-
ical studies on OCTOPAMINE2 receptors sub-types indicated that it
has not been found in vertebrates (Nathanson, 1985). Therefore, the
octopaminergic system in insect, in particular type-2 receptor is
a target for insecticidal action.
It is evident from various studies that some constituents such as
eugenol or thymol may work by blocking octopamine receptors
and/or by potentially working through the tyramine receptor
cascades (Enan, 2005a,b). Though cocaine has a fascinating and
complex medicinal history in human pharmacopeia. The insecti-
cidal effects of cocaine resulted from a potentiation of insect
octopaminergic neurotransmission (Nathanson et al., 1993). The
physical effects such as membrane disruption or blockage of the
tracheal systems may also be involved, however, conclusive
evidence is still lacking. Likewise, insecticidal properties of eugenol
have been reported via a low-dose activation of octopamine
receptors, which did not affect the response to octopamine on DUM
neurons and foregut activity. These results suggested a different
sub-type of octopamine receptor (Price and Berry, 2006).
3. Phytochemical sources and insecticidal activity
Before the discovery of modern pesticides, plant extracts con-
taining nicotine and pyrethrin were widely used in agriculture as
insecticides. Search for natural alternatives have improved the
development of bench top bioassays, an array of synthetic
approaches for known and novel natural products that shows
relevant activities such as feeding suppressors and deterrents
(Addor, 1994). Most of the secondary metabolites such as
terpenoids and alkaloids are reported as candidates for insecticidal
compounds that could be an effective alternative for insect pest
management. In fact, humans have used these for thousands of
years as dyes (e.g. indigo, shikonin), flavours (e.g. vanillin, capsa-
icin), fragrances (e.g. essential oils of rose, lavender), stimulants (e.
g. caffeine, nicotine), hallucinogens (e.g. morphine, tetrahydro-
cannabinol), poisons (e.g. strychnine, coniine) and medicines (e.g.
quinine, atropine). The plants secondary metabolites (such as sla-
vanoids, terpenes phenols, alkaloids, sterols, waxes, fats, tannins,
sugars, gums, suberins, resin acids carotenoids etc.) defend them
against microbial pathogens and invertebrate pests (Gottlieb, 1990;
Wink and Schimmer, 1999). The essential oils from various plants
are toxic to different insect pests viz. Artemisia judaica has anti-
feedant activity against Spodoptera littoralis (Abdelgaleil et al.,
2008), Nigella sativa against Callosobruchus chinensis (Chaubey,
2008), ginger against D. melanogaster (Xu et al., 2007), Eucalyptus
tereticornis against mature Anopheles stephensi (Nathan et al.,
2008), Litsea pungens and Listea cubeba against Trichoplusia ni
(Jiang et al., 2009), Chloroxylon swietenia against Anopheles gam-
biae, Culex quinquefasciatus and Aedes aegypti (Kiran and Devi,
2007), Calocedrus decurrens and Juniperus occidentalis against
adult A. aegypti, Xenopsylla cheopis and Ixodes scapularis (Dolan
et al., 2007), Chenopodium ambrosioides against green house
insect pests i.e. Planococcus citri, Frankliniella occidentalis (Cloyd
and Chiasson, 2007), rosemary against Agriotes obscurus
(Waliwitiya et al., 2005) etc. A review on botanicals oils by
Regnault-Roger (1997) highlights the complexity, ecological and
economical effects, potentials and trends in insect pest manage-
ment. Besides their role in the chemical defense of a plant, they
may also act as chemical signals to attract beneficial animals for
pollination and seed dispersal e.g. anthocyanins and monoterpenes
act as insect attractants in flowers, but may be insecticidal and
antimicrobial when present in leaves (Wink, 1999). In non-target
species, however, many of the compounds exhibit certain useful
biological activities. These can be grouped into three main chemical
classes: alkaloids, phenolics, and terpenes (terpenoids).
3.1. Alkaloids
These compounds are insecticidal at low concentrations and
also toxic to vertebrates. Their mode of action varies, but many
affect acetylcholine receptors in the nervous system (e.g. nicotine)
or membrane sodium channels of nerves (e.g. veratrin). Insecticidal
examples include nicotine (Nicotinia spp.), anabasine (Anabasis
aphylla), veratrin (Schoenocaulon officinale) and ryanodine (Ryania
speciosa). Physostigmine servs as a model compound for the
development of the carbamate insecticides, which is an alkaloid
isolated from the calabar bean, Physostigma venenosum (Stedman
and Barger, 1925). These chemicals are not volatile, and may be
used as repellents by burning plant material, either on a fire or in
a mosquito coil to create an insecticidal smoke that repels the
insects through direct toxicity. Alkaloids are found in large quan-
tities in many members of the Berberidaceae, Fabaceae, Solanaceae,
and Ranunculaceae families, all of which are used extensively as
traditional insect repellents (Secoy and Smith, 1983; Johnson,
1998).
3.2. Phenolics
Phenolic compounds are characterized by the presence of
a hydroxyl (eOH) group, attached to a benzene ring or to other
complex aromatic ring structures e.g. catechol, resorcinol, hydro-
quinone, pyrogallol etc. They range from simple phenol (MW 94,
found in essential oil of Pinus sylvestris) to polyphenols such as
anthocyanin pigments (MW 2000) and tannins (MW up to 20,000).
 R.S. Rattan / Crop Protection 29 (2010) 913e920
Hydrolysable tannins of oak are well-known as phenolics, which
can negatively influence the growth of the gypsy moth (Rossiter
et al., 1988). Plant phenolics have been regarded as one of the
important defenses against insects in various studies (Berbehenn
and Martin, 1994; Berbehenn et al., 1996; Henn, 1997). However,
their specific mode of action is not yet clearly known.
3.3. Terpenes (terpenoids)
Terpenes are widely used in the food, pharmaceutical and
perfume sectors, as well as in a wide range of pharmacological
applications. Terpenes are the largest group of natural products
from plants comprising essential oils, flavours, fragrances, and
lipid-soluble plant pigments. These hydrophobic compounds are
usually stored in plants in resin ducts, oil cells or glandular
trichomes (Wink and Schimmer, 1999). They are derived from 5-
carbon isoprene units [CH2]C(CH3)eCH]CH2], such as C5 hemi-
terpenes, C10 monoterpenes, C15 sesquiterpenes, C20 diterpenes, C25
sesterpenes, C30 triterpenes, C40 tetraterpenes, and C50 band over
polyterpenes. Examples of monoterpenes include limonene in
turpentines from some Pinus spp.., a-terpinene from P. sylvestris,
terpinolene in some Pinus turpentines, b-phellandrene in Pinus
contorta turpentine, and a- and b-pinene, camphene and 3-carene
that are abundant in turpentines from coniferous woods (Dev,
1989). The composition of turpentines is highly variable amongst
plant species, and some exhibit considerable toxicity to insects with
low mammalian toxicity (Fengel and Wegener, 1984; Golob et al.,
1999). The monoterpenes from the plant Porophyllum gracile are
shown to increase the toxicity of polyacetylene to lepidopteran
Ostrinia nubilalis. However, mixtures of terpenes containing
compounds with different physical properties may be more toxic
with longer persistence of defenses. Terpenes synergize the effects
of other toxins by acting as solvents to facilitate their passage
through membranes. An example of such synergism seems to occur
in conifer resin, which is a mixture of monoterpene olefins with
anti-herbivore and anti-pathogen activity, and diterpene that are
toxic and deterrent to herbivores. A well-studied example is the
pyrethroids (i.e. monoterpene esters) found in certain Chrysan-
themum species leaves and flowers. The toxin causes disturbance in
the nervous system leading to paralysis and mortality, lead to
development of the most successful commercial pesticides (Raffa
and Priester, 1985; Gershenzon and Croteau, 1991).
4. Development of resistance vs phytochemicals
The chemical defenses of plants are often more subtle in their
action on herbivores, which has been shaped in the evolutionary
process of 300 million years (Isman, 1996). However, natural
products are not immune to herbivore resistance, although, it has
been suggested that extracts from plants, which contain numerous
compounds, are more complex in comparison to synthetic pesti-
cides and therefore, delays the build up of resistance (Völlinger,
1987). In a laboratory study, Plutella xylostella (L.) develops resis-
tance rather quickly to all major groups of synthetic pesticides.
However, it was unsuccessful to develop resistance to Neem after
42 generations of selection, which is attributed to Neem constitu-
ents and complex mode of action (Völlinger, 1987; Schmutterer,
1988). The combination of behavioral and physiological actions of
botanical pesticides deters the development of resistance (Rice,
1993). This is common phenomenon in natural ecosystem where
herbivorous insects are controlled by plant allelochemicals.
However, farmers often use large quantities of synthetic insecti-
cides to control herbivorous insects, which has led to development
of resistance e.g. frequent sprays to manage P. xylostella coupled
with rapid turnover of generations in the tropical climate, has
917
resulted in the development of resistance practically to all cate-
gories of synthetic insecticides including Bacillus thuringiensis in
the field (Sun et al., 1986; Miyata et al., 1986; Saito et al., 1995; Ferré
and van Rie, 2002). If this trend continues in the future, more
insecticides and money will be used for the management of this
pest and other agricultural insect pests, which are vulnerable to
agricultural production. Therefore, insecticides with novel mode of
action are often sought regularly as a mean to cope with the
resistance problems. Though, the presence of multiple mechanisms
of resistance (i.e. acetylcholinesterase insensitivity, reduced pene-
tration, nerve insensitivity and detoxification of insecticides)
suggests that resistance may occurs to any class of insecticide given
enough time, consistent selection pressure, and a large enough
population for selection to occur (Sun, 1990). There are less chances
of resistance development with novel molecules from plant origin
with specific or multiple target sites. Unexplored phytochemicals
may have unknown site of action, which are selective to insects
and, are safe to humans and environment.
5. Future perspectives
Sustainable growth in agriculture is crucial for most of the
developing countries to sustain the growing populations. Synthetic
crop protection chemicals are associated with pest resurgence,
impact on non-target organisms, health and environment. Hence,
there is need to develop safe alternative crop protectants (insecti-
cides), which are more specific with wide range of activities.
Identification of novel effective insecticidal compounds is essential
to combat impacts of synthetic pesticides. An increasing number of
researchers are reconsidering botanicals containing active phyto-
chemicals in their efforts to address some of these problems.
Consequently, more target-selective and biodegradable
compounds are needed to replace the environmentally persistent
chemicals having broad-spectrum toxicity, which may play an
important role in sustainable pest management (Alkofahi et al.,
1989; Arnanson et al., 1992; Isman, 2006).
The multiple mechanisms of action give phytochemicals unique
properties that make it very useful in today’s agricultural industry
e.g. azadirachtin (Copping and Menn, 2000). The development of
imidacloprid and a series of neonicotinoids is probably one of the
greatest landmarks in the history of pesticide chemistry. In the past
few years, neonicotenoids have been established as key insect pest
management agents viz. thiamethoxam etc. Likewise, the knowl-
edge of human b3-GABA receptors, which pharmacologically
resemble insect GABA receptors, were used for exhaustive site-
directed mutagenesis and ligand docking simulations, leading to
novel conclusion that structurally diverse antagonists fit the same
site in the receptor channel pore (Casida and Tomizawa, 2008). The
biogenic amines have also been implicated in a variety of physio-
logical processes in invertebrates (Hirashima, 2008). Octopamine
and tyramine are distinguished only by the presence or absence of
a hydroxyl group at the b-position (Ozoe and Huang, 2008). These
structurally closely related amines regulate levels of intracellular
cAMP in opposite directions, i.e. up- and down-regulation, by
acting at distinct G-protein coupled receptors. Based on the
assessment of about 100 compounds for endocrine disruptors
activity using a reporter gene assay, three-dimensional quantitative
structureeactivity relationships were analyzed to elicit responses
via androgen receptors (Tomura et al., 2008). The design of novel
crop protection chemicals possessing superior properties is of great
significance. Thus, the findings regarding the natural biomolecules
binding site structures would be helpful for designing of new
insecticidal molecules with novel mode of action. Further
enhancement of their activity and stability could be done by
918 R.S. Rattan / Crop Protection 29 (2010) 913e920
encapsulation (Pillmoor et al., 1993; Lee et al., 2005; Kumari et al.,
2010).
The knowledge on the mode of action of available insecticides
will help to determine the chemical properties of novel compounds
that may be ideally suited for modification of insect behaviour at
doses that can be used safely and economically in agriculture
(Haynes, 1988). The co-evolution of plants along with insects has
compelled the use of natural chemical defenses for the manage-
ment of insect pests. Since, botanicals are less likely to cause
ecological damage, a large number of plants have been tapped for
their insecticidal properties against various insect pests (Ryan and
Byrne, 1988). Apart from all the advantages and safety aspects of
botanicals as compared to synthetics, it will also give farmers the
necessary psychological satisfaction. At the same time, it will be
helpful in reducing the present excessive use of synthetic insecti-
cides, which are not compatible with many biological and microbial
components of an IPM package. Phytochemicals can serve as model
compound for the development of chemically synthesized deriva-
tives with enhanced activity or environmental friendliness
(Chitwood, 2002; Park et al., 2005). However, their functions and
clarity on specificity of the metabolites responsible for proclaimed
insecticidal activity is lacking. The current view on mode of action is
still rather linear, whereas, it is likely that in reality mechanism will
behave like a web, in which pathways and molecules will interact in
a flexible and dynamic way. The plant origin insecticides could be
exploited for the development of novel molecules with highly
precise target for sustainable insect pest management in
agriculture.
Acknowledgements
Author gratefully acknowledge the Director, Institute of Hima-
layan Bioresource Technology (CSIR), for encouragement and
guidance. IHBT publication number of this manuscript is 1073. RSR
acknowledges the help rendered by Dr Ruhi in improving the
manuscript.
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