01-11-2014

Biopesticides with special

reference to Bt toxin

A Pest

PEST = any organism (bacteria, fungi, plant,
animal) that interferes in some way with human
welfare or activities.

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Losses caused by different Pests

Rodents & Others

12%
Weeds
Diseases

37%

22%

Insects 29%
On an average 18% of the crop yield is lost due to pests
(Annual monetary loss: Rs.60,000 Crores)
Source: Working Subgroup on Plant Protection, Planning Commission, India

Economic loss from pest insects is due to:

Reduced yields
Lowered quality of produce
Increased costs of production and harvesting
Expenditures for control

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Pest management methods

PESTICIDE = toxic chemicals used to reduce the

size of and control the pest population.

Grouped by their target organism
Insecticide, Herbicide,
Fungicide, Rodenticide

85% of pesticides worldwide used for Agriculture.

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Concerns regarding use of chemical pesticides

Development of pesticide resistance - insects and
weeds.(insect resistance -nearly 200 species).

Disrupt the natural ecosystem and natural biodiversity.
Hazardous effect on the non-target, useful arthropods. They
kill beneficial insects and plants (non selective).

Pollution runoff of herbicides and insecticides into
irrigation water and then into rivers - damages wildlife
habitat, kills fish.

Reappearance of residues in things of human use, Residues
on crops and soil

Cause cancer organophosphates

Bioaccumulation

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Bio-Pesticides

Bio-pesticides are certain types of pesticides

derived from natural material such as animals,
plants, bacteria, and certain minerals.
Many bio-pesticides are less toxic and pose a
lower risk than conventional pesticides which
can be quite toxic.

Biopesticides: mass produced biologically based agents

used for the control of plant pests
Living organisms (natural enemies)
Micro-organisms
(Arthropods & nematodes)

Naturally occurring substances -biochemicals

Plant extracts.
Semiochemicals (pheromones & allelochemicals).
Commodity substances.

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Micro-organisms use in biocontrol

Fungicide, 35

Insecticide, 50
Herbicide, 12

Acaricide, 1
Fungicide, 35
Fungicide and nematicide, 1
Herbicide, 12
Insecticide and acaricide, 1
Nematicide, 5

Bactericide, 2
Fungicide and bactericide, 2
Fungicide and plant growth regulator, 1
Insecticide, 50
Molluscicide
Plant growth promoter, 1
(Copping, 2004)

Biopesticides

Advantages:
Lower potency than synthetic pesticides
Low toxicity to non-target organisms
Little or no residue: Fast action and breakdown so low
environmental impact
High selectivity: Often very specific
Compatible with other control agents
Inexpensive to develop
Natural enemies used in ecologically-based IPM

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Bio-pesticides
Limit:
Contact products so adequate coverage is essential to
have a good efficacy.
Often time consuming

Biological Weed Control

Principle is that an insect or disease selectively
targets a weed without endangering nontarget plants.
Insects released to control weeds include
natural enemies lantana, parthenium, water
hyacinth, etc. have kept these weeds at
reduced levels, but has not resulted in
elimination.
Mycoherbicides
Phytotoxins from plants

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Fate of parthenium after timely augmentation of the beetle ,

Zygogramma bicolorata

Use of Biopesticides in Weed Management

Bioherbicide : herbicides based on any living organism.
Indigenous bioherbicides are considered favourable alternatives to
assess chemical herbicide issue relating to human and animal poisoning,
residues, environmental pollution and weed resistance.
Present day bioherbicides include:
Phytopathogens
Phytotoxins from microbes
Phytotoxins from plants
Semiochemicals

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Microbial management of weed

These days several microbes (including
fungi, bacteria, viruses and nematodes) are
being used to control weeds.
Mycoherbicide:

bioherbicide

based

on

fungal formulation
Some

of

the

commercially

available

mycoherbicide include:
Commercial

Pathogen

Target weed

ABG5003

Cercospora rodmani

Water hyacinth

BioChon

Chondrostereum purpureum

Prunus serotina

DeVine

Phytophthora palmivora

Milkweed vine

Colletotrichum gloeosporioides

Malva pusilla

Xanthomonas campestris

Zoysia tenuifolia

name

BioMal
Camperico

Stumpout: A
commercially
available
mycoherbicide

Effect of Alternaria alternata on waterhyacinth

Before

After 50 days

Diseased leaves of Waterhyacinth

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Phytotoxins from microbes

Naturally occurring toxic chemicals produced by microbes for various
purpose like:
attack or gain access to host cells by helping in dissolving cell
membranes
as protective measures against encroaching organisms
Mycotoxins obtained from plant pathogenic fungi are the most effective
biologically based alternatives to chemical herbicides.
These phytotoxins belong to a wide array of chemical substances
including sesquiterpenoids, sesterterpenoids, diketopiperazines, peptides,
spirocyclic lactams, isocoumarins, and polyketides.

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Mycotoxin

Pathogen

Target weed

AAL-Toxin

Alternaria alternata

Jimson weed

Colletotrichin

Colletotrichum sp.

Solanum sp.

Curvulin

Drechslera indica

Spiny amaranth

Fusarium
Moniliformin

Spiny amaranth
moniliforme

Anisomysin

Streptomyces sp.

Barnyard grass

Fractions separated by
solvent extraction of cell free
culture filtrate of
Alternaria alternata

and crab grass

Effect of various fractions separated by

solvent extraction on waterhyacinth

Use of Biopesticides in Insect Pest Management

Bioinsecticide : insecticides based on any living organism.

Present day bioherbicides include:
Entomopathogens
Entomotoxins from microbes
Entomotoxins from plants
Semiochemicals- pheromone

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Microbial control of insects

Microbial
insecticides
are
comprised
of
microscopic living organisms (viruses, bacteria,
fungi, protozoa, or nematodes) or the toxins
produced by these organisms.
They are formulated to be applied as conventional
insecticidal sprays, dusts, liquid drenches, liquid
concentrates, wettable powders, or granules.
Each product's specific properties determine the
ways in which it can be used most effectively.

Entomopathogenic Nematodes
 Some entomogenous nematodes have characteristics that allow them to be
considered with the pathogens.
 The most important insect pathogenic nematodes for biological control are very
small and use mutualistic bacteria to kill the host.

Eutectona machaeralis, pest of teak

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Entomopathogenic Nematodes
Although nematode species in at least 20 families are primary or facultative
parasites of insects, those in the order Rhabditida have been most exploited as
biological control agents.
 Species in the genera Steinernema and Heterorhabditis (Steinernematidae and
Heterorhabditae, respectively), are particularly amenable to mass production and
application in a variety of pest systems.
 Entomopathogenic nematodes enter the host via natural body openings or
through the cuticle.
 Some species utilize an anterior stylet or a tooth to rasp the cuticle and gain
entrance into the hemocoel.
 Others ingress by ovipositing on the host food source and the eggs hatch in the
host midgut.
 Effects of nematode parasitism on the hosts can be sterility, reduced fecundity,
reduced mobility and life span, behavioural and morphological changes, and death.

Entomopathogenic Nematodes
The non-feeding, developmentally arrested infective juvenile seeks out
insect hosts and initiates infections.
When a host has been located, the nematodes penetrate into the insect
body cavity, usually via natural body openings (mouth, anus, spiracles) or
areas of thin cuticle.
Once in the body cavity, a symbiotic bacterium (Xenorhabdus for
steinernematids, Photorhabdus for heterorhabditids) is released from the
nematode gut, which multiplies rapidly and causes rapid insect death.
The nematodes feed upon the bacteria and liquefying host, and mature
into adults. Steinernematid infective juveniles may become males or
females, where as heterorhabditids develop into self-fertilizing
hermaphrodites although subsequent generations within a host produce
males and females as well.

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Entomopathogenic Nematodes

Entomopathogenic Nematodes
The life cycle is completed in a few days, and hundreds of
thousands of new infective juveniles emerge in search of fresh
hosts.
Thus, entomopathogenic nematodes are a nematode-bacterium
complex.
The nematode may appear as little more than a biological syringe
for its bacterial partner, yet the relationship between these
organisms is one of classic mutualism.
Nematode growth and reproduction depend upon conditions
established in the host cadaver by the bacterium.
The bacterium further contributes anti-immune proteins to assist
the nematode in overcoming host defenses, and anti-microbials
that suppress colonization of the cadaver by competing secondary
invaders.
Conversely, the bacterium lacks invasive powers and is
dependent upon the nematode to locate and penetrate suitable
hosts.

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01-11-2014

Insects infected with Steinernema

nematodes are usually light tan in
color.
Note the adults (larger nematodes)
and the infective juveniles (the tiny
nematodes forming a cloud around the
grub.

Insects infected with

Heterorhabditis
nematodes are usually
a reddish color.

Entomopathogenic Protozoa
 Of some 14,000 described species of Protozoa, about 500 are pathogens of
insects. Many are chronic pathogens that may debilitate a host without producing
obvious disease symptoms but some species are extremely virulent, causing
stunted growth, slow development, and early death.
 Entry into the host is typically by ingestion, but some can invade through the
cuticle.
 Some species may be transovarially transmitted from infected females to their
offspring.
 Species that invade the cells of the host are usually found in the cell cytoplasm
and are typically more pathogenic than extracellular species.
 Some protozoans exhibit tissue tropism, infecting only certain tissues or organs,
others are systemic.
No toxins have been found to be associated with protozoa in insects.
 Death or debilitation of infected hosts may be, for example, the result of
competition for metabolites, disruption of normal cell and tissue function, or
blockage of the gut or other organs by extracellular species.
 The insect-pathogenic Protozoa are currently recorded from four major groups:
Amoebas, Gregarines, Flagellates and Ciliates.

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Insect viruses
Viral diseases have been found in 13 insect orders and most likely occur in all
orders. Viruses that are primarily or exclusively found in insects are currently
placed in 12 families (Miller, 1998):

DNA Viruses: Baculoviruses (Nuclear polyhedrosis viruses- NPV and
Granuloviruses-GV), Ascoviruses, Iridoviruses, Parvoviruses, Polydnaviruses
and Poxviruses.

RNA Viruses: Reoviruses (Cytoplasmic polyhedrosis viruses), Nodaviruses,
Picorna-like viruses and Tetraviruses.

they do not infect vertebrates, nonarthropod invertebrates, microorganisms or
plants.

Baculoviruses as Biocontrol Agents

Baculoviruses are rod-shaped, double stranded DNA viruses
that can infect and kill a large number of different invertebrate
organisms
Immature (larval) forms of moth species are the most common
hosts, but these viruses have also been found infecting
sawflies, mosquitoes, and shrimp.
Baculoviruses have limited host ranges and generally do not
allow for insect resistance to develop
Slow killing of target insects occurs
In order to speed killing (enhance effectiveness), several genes
can be expressed in the baculovirus including diuretic
hormone, juvenile hormone esterase, Bt toxin, scorpion toxin,
mite toxin, wasp toxin, and a neurotoxin (see Table 16.7)

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Electron Micrograph of a
Baculovirus Occlusion Body

17

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Table 16.7

Fungal pathogens of insects

 Entomopathogenic fungi are able to invade their insect hosts by
penetrating directly through the cuticle.
The fungal spore first adheres to the cuticle.
 Under appropriate conditions the spore germinates, penetrates the
cuticle of the host and enters the hemocoel.
 Fungal reproduction occurs in the hemocoel of the insect host.
 As the hemocoel becomes filled with hyphal bodies, the insect
usually dies and the fungus continues to develop saprophytically.
 After the body of the dead insect is filled with mycelia, fruiting
structures emerge from the cadaver and produce infectious spores.
 Dead insect has the consistency of a moist loaf of bread and,
depending on the colour of the spores or conidia, may appear white
or some darker colour.

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Fungal pathogens of insects

 Tanada and Kaya (1993) listed 8 classes, 13 orders and 57 genera that contain
entomopathogenic species of fungi.
 There are five major groups of fungi: the Flagellate fungi or Chytridiomycetes, the
Oomycetes (also flagellate but also not true fungi), the Zygomycetes, the
Ascomycetes, and the Basidiomycota.
 The Zygomycota and the Ascomycota contain common insect pathogens that are
also useful in biological control programs.


Fungal pathogens of insects

Examples of Entomopathogenic fungi are:
1. Metarrhizium anisopliae
2. Beauveria bassiana
3. Entomophthora spp.

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Entomopathogenic Fungi
Beauveria spp. "White" Fungus
Beauveria bassiana is called the white fungus of insects
because most of the strains produce external spores that
make the infected insect appear coated with a white powder
or cottony material.

Metarhizium spp. "Green" Fungus

Metarhizium is usually called the green fungus of insects.
The fungus has white mycelia within the body, but when it is
ready to form spores, the spores coat the killed host with a
velvety covering of olive-green.

A bluegrass billbug adult (above) and

Japanese beetle larva (right) infected with
Beauveria.

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01-11-2014

Green June beetle

grubs killed by
Metarhizium.

Japanese beetle grub

infected with
Metarhizium.

Bacterial pathogens of insects

They can be divided into two broad categories, nonspore-forming bacteria and spore-forming bacteria.
Although most of the species isolated from diseased
insects are non-spore-forming, spore-forming bacteria in
the genus Bacillus are the most important from the
standpoint of biological control since they are resistant to
environmental changes.
 Among the spore formers crystalliferous ones are better
than non-crystalliferous ones because of the toxic nature
of crystals they produce.

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Entomogenous Bacteria

Sporeformers

Obligate

Non-sporeformers

Facultative

Crystalliferous

Potential

Facultative

Ex

Non-crystaliferous

Serratia
Ex
Paenibacillus
popilliae

Clostridium sp.

Ex

Ex

B. thringensis

B. cereus

Ex

Pseudomonas sp.
Aerobacter sp.

Ex. 1: Milky disease bacterium, Paenibacillus popilliae, on

white grubs of Japanese beetles
There are numerous bacteria that can infect and kill insects.
The most common ones encountered in landscapes are the
ones that kill white grubs milky disease bacteria.
Spores of Paenibacillus popilliae (formerly Bacillus popilliae)
infect larvae of Japanese beetles (Popillia japonica).
The milky disease bacterium, Paenibacillus popilliae, causes
the blood of white grubs to turn a milky white color. Eventually,
the bacteria overwhelm the grub.
Though milky disease spores are commercially available,
there is little evidence that use of such products actually
increase infection over the long run.

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The adult Japanese beetles

Milky disease bacterium, Paenibacillus popilliae

It is a soil-dwelling, gram-positive, spore forming, rod-shaped
bacterium. It is responsible for a disease (commonly called milky
spore) of the white grubs of Japanese beetles.
Milky Spore in the soil is not harmful to beneficial insects, birds,
bees, pets or man
Milky Spore, like other bacteria, is highly survivable in cold and
drought conditions.

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Milky disease bacterium, Paenibacillus popilliae,

on white grubs of Japanese beetles
Spores, residing in the soil are ingested by beetle larvae
Day 2: Germinate in larval gut.
Day 3-5: Vegetative cells proliferate, attaining maximum
numbers.
Day 5-10: Some penetrate the gut wall and grows in the
hemolymph
Day 14-21: A few spores form larva develops the typical
milky appearance.
Host dies

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Normal grub (left) and a milky disease infected grub (right). Note color
of blood droplet where the tip of the leg was pinched off.

Resident spores in the soil are swallowed by grubs during their normal
pattern of feeding on roots. This ingestion of the spore by the host
activates reproduction of the bacteria inside the grub. Within 721 days
the grub will eventually die and as the grub decomposes, billions of
new spores are released into the soil.

Spores are ingested by Japanese beetle larvae (grubs)

Spores become active bacteria and multiply in the grubs,
which continue to live.
Prevents larval maturation.
When the larvaes bacterial population reaches a high
enough density, bacterial spores are released to the soil to
await ingestion by future beetle larvae.
Infected beetle larvae die when the spores are released.
Thus, they greatly decrease the numbers of grubs and adult
beetles, thereby reducing plant damage.
Spores also infect larvae of some closely related beetles

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Advantages
Very narrow host range (they are effective against
Japanese beetles, only)
Complete safety for man and other vertebrates
Compatibility with other control agents including
chemical insecticides

Disadvantages
High cost of production in vivo,
Slow rate of action
Lack of effect on adult Japanese beetles
Need for large areas to be treated for effect.

Ex. 2: Bacillus thuringiensis

A Short History of Bacillus Thuringiensis (BT)
Bt is a naturally occurring soil bacterium.
It was first detected in 1902 in the dying larvae
of Bombyx mori by Ishiwata,
he reported his finding in the book: "Pathology
of the Silkworm".
It was first isolated from the larvae of
Ephestia kuehniella by Berliner in 1911 after
he noted that it had the capacity to kill certain
insects in their larva stage.

Ephestia kuehniella

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Ex. 2: Bacillus thuringiensis

A Short History of Bacillus Thuringiensis (BT)
It was regarded as potential biocontrol agent by
Steinhaus in 1956.
1st commercial product containing this bacillus
was available in USA in 1958.
Natural Bt is highly specific, with toxicity
limited to only some species of one of the
major groups of insectstypically Lepidoptera
(butterflies/moths),
Coleoptera
(beetles),
hymenoptera (wasps, bees, ants) or Diptera
(flies/ mosquitoes).

Scientific classification of Bacillus thuringiensis

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Bacillus Thuringiensis (BT)

Bacillus thuringiensis is a gram-positive, spore-forming
bacterium which, during sporulation, produces protein
crystals (CRY). It is characterized as a widespread insect
pathogen, and its insecticidal activity is attributed to the
parasporal crystals.
A variety of strains have been isolated from different
habitats and, to date, more than 100 crystal protein
genes have been sequenced.

Bacillus Thuringiensis (BT)

The toxicity of these crystal
proteins against certain insects
and their high specificity led to the
development of bio-insecticides
for the control of pest insect
species
among
the
orders
Lepidoptera,
Diptera,
and
Coleoptera.

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Characteristics of BT
Bt subspecies can synthesize more than one parasporal inclusion.
The parasporal inclusions are formed by different insecticidal
crystal proteins (ICP).
The crystals have various shapes (bipyramidal, cuboidal, flat,
rhomboid, spherical or composite with crystal types), depending on
their ICP composition.
During sporulation many Bt strains produce crystal proteins
(proteinaceous inclusions), called Cry proteins which are encoded
by cry genes, and have insecticidal action. This has lead to their use
as insecticides and more recently to genetically modified crops
using Bt genes.
In most strains of B. thuringiensis the cry genes are located on the
plasmid.
Different domains of the ICP are responsible for host susceptibility
(receptor recognition) and toxicity (pore formation).

Insecticidal toxin of Bacillus thuringiensis

A bacterial insecticide either produces a toxic
substance that kills an insect species or has the
capability of fatally infecting a specific target
insect.
The most studied, most effective, most often
utilized microbial insecticides are the toxins
synthesized by Bacillus thuringiensis.
This bacterium comprises a number of different
strains each of which produces a different toxin
that can kill certain specific insects.

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For ex.
B. thuringiensis kurstaki is toxic to: lepidopteran
larvae of moths and butterflies (skippers, cabbage
worm and spruce budworm).
B. thuringiensis israelensis kills Diptera such as
mosquitoes and black flies.
B. thuringiensis tenebrionis is effective against
Coleoptera (beetles) such as the potato beetle and
the boll weevil.
Other B. thuringiensis strains with different toxins
that are specific toward certain insects.

Mode of action
To kill an insect pest, B. thuringiensis must be
ingested as the contact of the bacterium or
the toxin with the surface of an insect has no
effect on the target organism.
B. thuringiensis is generally applied by
spraying, so it is usually formulated with
insect attractants to increase the probability
that the target insect will ingest the toxin.

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How Does it Work

The insecticidal activity of B. thuringiensis kurstaki
and other strains is contained within a very large
structure called the parasporal crystal, which is
synthesized during bacterial sporulation.
The crystal is an aggregate of one kind of protein
that can be dissociated by mild alkali treatment to
yield two subunits of 130 kDa each.
The parasporal crystal is not the active form of the
insecticide; rather, it is a protoxin, a precursor of
the active toxin.

When the para-sporal crystal is ingested by a target

insect, the protoxin is activated within its gut by
the combination of alkaline pH (7.5 to 8.0) and
specific digestive proteases, which converts the
protoxin into an active form with 68 kDa.
When the toxin changes to its active form, it
inserts itself into the membrane of the gut
epithelial cells of the insect and creates an ion
channel through which excessive loss of cellular
ATP occurs.
About 15 min after the channels forms cellular
metabolism ceases, the insect stops feeding,
becomes dehydrated, and eventually dies.

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Two things make this process very specific;

1. The need for an alkaline media (pH 9 to 12)
2. The need for specific proteases

B. thurigienis kurstaki is applied by spraying

approximately
1 .3 to 2.6 X 108 spores per square foot of the
target area at the peak of the larval population
of the target organism.
The crystal is short lived as it breaks down after
exposure to sunlight so it is appropriate to spray
it in cloudy days.

How BT works?

BT SPORES

Normal gut bacteria

BT crystalline Toxin 200px.

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01-11-2014

Protoxin is cleaved by a gut protease to produce an

active 60 kDa toxin: Delta-endotoxin.
Binds to the midgut epithelial cells
Creates pores in cell membranes & leads to
equilibration of ions
Gut is rapidly immobilised & the epithelial cells lyse
Larvae stop feeding
Gut pH is lowered by equilibration with the blood
pH.
Lower pH enables the bacterial spores to germinate
The bacteria invade the host, causing a lethal
septicaemia.

How BT works?
Mode of Action

1.

2.
3.

4.
5.

The Bacillus thuringiensis is only effective when eaten by

specific family of insects with a specific (usually alkaline) gut
pH and the specific gut membrane structures required to bind
the toxin. (typically butterflies, moths, beetles, flies and
mosquitoes).
Not only must the insect have the correct and be at a
susceptible stage of development, but the bacterium must be
eaten in sufficient quantity.
When ingested by a susceptible insect, the spores feed on natural
intestinal flora then it burst releasing the protein toxin
(Crystalline protein) damaging the gut lining (the intestinal
walls), leading to a kind of leaky gut condition.
Affected insects stop feeding and die from the combined
effects of starvation, tissue damage and gastrointestinal
infections by other pathogens like bacteria and funguses.
The natural Bt spores do not usually spread to other insects
or cause disease outbreaks on their own as occurs with many
pathogens.

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Figure 16.1

B. thuringienis parasporal crystal composed of Cry1

protoxin protein. Conversion of the 130-kDa protoxin
into an active 68-kKa toxin requires an alkaline
environment (pH 7.5 to 8) and the action of a specific
protease, both of which are found in the insect gut.
The activated toxin binds to protein receptors on the
insect gut epithelial cells.

Figure 16.3

The toxin is inserted in gut epithelial cell membranes of the

insect and forms an ion channel between the cell cytoplasm
and the external environment, leading to loss of cellular ATP
and insect death.

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01-11-2014

Advantage:
Because for the toxin to be effective it has to be
ingested, this limits the susceptibility of non-target
insects and other animals to this insecticide.

Drawbacks:
Insects that attack plant roots are less likely to ingest
a B. thuringiensis toxin that has been sprayed on the
surface of a host plant.
B. thuringiensis toxin can only kill a susceptible insect
during a specific developmental stage.
It costs 1.5-3 times as much as chemical insecticides
Resistance of insects to the toxins produced by these
bacteria might occur.

Bt toxins and their classification

Bt produces 2 types of toxin
Cry (crystal) toxins, encoded by cry genes (> 50 genes !!!!)
Cyt (cytolytic) toxins, to augment Cry toxins

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Strain development
Cry toxins are encoded by genes on 5-6 different
plasmids of Bt
A sea of combinations & Cry toxins why?
Plasmids can be exchanged between Bt strains by a
conjugation-like process
Bt contains transposons (transposable genetic elements
that flank genes and that can be excised from one part of
the genome and inserted elsewhere)

So, commercially, genetically engineered strains with

novel toxin combinations

Plants genetically engineered with Bt gene

Genetically engineering to contain the deltaendotoxin gene from Bt
Bt corn
Bt potato
Bt cotton
Bt soybean
The "downside
Perpetual exposure of insects to toxins
Creates a very strong selection pressure for the
development of resistance to the toxins.

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Advantages in expressing Bt toxins in

transgenic Bt crops:
Level of toxin expression can be very high thus
delivering sufficient dosage to the pest
Toxin expression is contained within the plant
system and hence only those insects that feed on
the crop perish
Toxin expression can be modulated by using
tissue-specific promoters
Replaces the use of synthetic pesticides in the
environment.

In an industrial scale
Produced in controlled fermentor in deep
tanks of sterilized nutrient liquid medium
Endotoxins & living spores are harvested as
water dispersible liquid concentrates for
subsequent formulation.

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Example of using Bacterial insecticides;

Bacillus thuringiensis sub sp. kurstaki
was used as major means of
controlling spruce budworm in Canada.
Its use was increased from 1% in 1979
to around 74% in 1986 for treating
spruce budworm in Canada
In other countries, B. thuringiensis
kurstaki has been used against tent
caterpillars, gypsy moths, cabbage
worms, cabbage loopers, and tobacco
hornworms.

NOTES:
Technically, the bacterium, Bacillus thuringiensis (Bt), is not a
true biological control. This is because the bacterium does
not cause an infection within the insect. The bacteria actually
grows in soils. However, many strains of this bacterium
contain a protein toxin crystal that is released if the bacterium
is digested. The protein toxin destroys the insect gut lining
which causes a secondary infection or starving of the insect.
Many strains of Bt are known and only a few have been found
to have insect killing properties.

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Bacillus thuringiensis
Bt -endotoxins
Variety

Target

Bt. kurstaki
Bt. aizawai
Bt. israelensis
Bt. tenebrionis
(=san diego)
Bt. japonensis
(strain 'Buibui')

Examples

caterpillars
caterpillars
mosquitoes
leaf beetles

Dipel, MVP
Mattch
Vectobac
M-one

scarab grubs

M-press

Target insects for Bt toxin

Cry toxins have specific activities against insect species of the orders Lepidoptera
(moths and butterflies), Diptera (flies and mosquitoes), Coleoptera (beetles),
Hymenoptera (wasps, bees, ants and sawflies) and nematodes.

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Insecticidal toxin of Bacillus thuringiensis

Current market for pesticides is $30 billion/year
Several different strains and subspecies of B.
thuringiensis exist and produce different toxins that
kill specific insects (Table 16.1)
Used as alternative to DDT and organophosphates
since the 1920s
Bt toxin is used as specific insecticides under trade
names such as Dipel and Thuricide

Table 16.1 Some properties of the insecticidal toxins

from various strains of B. thuringiensis
Strain/subsp.

Protein size

Target Insects

Cry #

berliner

130-140 kDa

Lepidoptera

CryI

kurstaki KTP, HD1

130-140 kDa

Lepidoptera

CryI

entomocidus 6.01

130-140 kDa

Lepidoptera

CryI

aizawai 7.29

130-140 kDa

Lepidoptera

CryI

aizawai IC 1

135 kDa

Lepidoptera, Diptera

CryII

kurstaki HD-1

71 kDa

Lepidoptera, Diptera

CryII

tenebrionis (sd)

66-73 kDa

Coleoptera

CryIII

morrisoni PG14

125-145 kDa

Diptera

CryIV

israelensis

68 kDa

Diptera

CryIV

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3. Bacillus sphaericus
Gram-positive bacterium
Used primarily as a larvicide
An obligate aerobe bacterium used as a larvicide for
mosquito control
Forms spherical endospores
Can be isolated from soil, leaf surfaces and aquatic
systems
Produces a 100 kDa protein that acts as a larvicidal
toxin.
Highly effective against the larva of the Wyeomyia
mosquitoes, drastically reducing their population.

Bacillus sphaericus

Effective against Culex spp.

Larvicides are more effective and less toxic
than adult mosquito sprays
Unlikely to result in human exposure

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Mode of action
B. sphaericus spores are eaten by mosquito
larvae
Toxins released into the mosquito's gut
Larvae stop eating
Effective against actively feeding larvae, and
does not affect mosquito pupae or adults.

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Constraints of Cotton Production

Change in pest Scenario
a. Excessive and indiscriminate use of pesticides
b. Improper choice, quantity and application of pesticides.
c. Use of pesticide mixtures

(Has lead to)

a. Resurgence of minor pests
b. Resistance to pesticides
c. Increase in cost of protection

Cotton also carries environmental controversy, particularly in the developing

world, where dangerous pesticides are heavily employed.
Pesticides compromise as much as 50% of the costs of cultivation for many
farmers and the increasing use of pesticides has led to high levels of debt and
bankruptcy among poorer farmers
An alternative to increased pesticide use is the encouragement of natural
predators of the bollworm, including ladybirds, lacewings, damsel bugs, night
stalking spiders, common brown earwigs, fire ants, muc wasps, and the
trichogramma wasp.
Pest traps, spot treatment and organic pesticides are also more sustainable
alternatives.
A more common alternative today is to turn to genetically-modified crops
produced by companies such as Monsanto. The most common species, Bt or
Bacillus thuringiensis, is advertised as fully resistant to the common bollworm.
Early use shows that this is true, as such large portions of cotton production in
the US and China (20 and 30% respectively) now use Bt strains.
Bt Cotton seeds were introduced by Bollgard Cotton, a trademark of the
Monsanto group. Bt Cotton was first introduced to the U.S. in 1996

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Bt Cotton is produced by inserting a synthetic version of a

gene from the naturally occurring soil bacterium Bacillus
thuringiensis, into cotton.
The primary reason this is done is to induce the plant to
produce its own Bt toxin to destroy the bollworm, a major
cotton pest.
The gene causes the production of Bt toxin in all parts of the
cotton plant throughout its entire life span. When the
bollworm ingests any part of the plant, the Bt cotton toxin
pierces its small intestine and kills the insect.

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