Lecture no: 26

Biotechnological approaches in pest management

The

European

federation

of

biotechnology

in

1982

defined

biotechnology as the integrated application of biochemical, microbiological

and

engineering

sciences

to

the

technological

employment

of

microorganisms, cell cultures or their components.

Biotechnological approaches canbe utilised in the following areas in pest
management:
i) Genetic improvement of entomopathogens
ii) Genetic improvement of entomophages
iii) Development of trangenic plants
iv) Genetic control of insect pests
v) Use of tissue culture and fermentation technology
a) Genetic improvement of Baculovirus
b) In vitro production of Baculovirus
c) Genetic engineering of Baculovirus
vi) Wide hybridization
Use of molecular biology techniques for the management of insect
pests. The following are some strategies.
1.Wide hybridization: This technique involves transfer of genes from one
species to other by conventional breeding.
transferred from a different species.

The genes for resistance are

e.g. WBPH resistant gene has been

transferred to Oryza sativa from O.officinalis.

2.Somaclonal variability
Somaclonal variation refers to variation observed in tissue culture
derived progenies. A general term somaclone has been proposed for plants

derived from any form of cell culture and the variation among such plants
termed as somaclonal variability.
Somaclonal variants of sugarcane and sorghum have been obtained with
good levels of resistance to
1. Sugarcane borer, Diatraea saccharalis and
2. Cut worm, Spodoptera litura
3.Transgenic plants or genetically engineered or modified plants:
Transgenic plants are plants which possess one or more additional genes.
This is achieved by cloning additional genes into the plant genome by
genetic engineering techniques. The added genes impart resistance to pests.
These transgenic plants produce insecticidal or antifeedant proteins
continuously in the plants under field conditions. In this approach,
identification of useful genes to be transferred is a limiting factor. Because of
this reason, the genes conferring insect resistance transferred by this
approach are mostly limited to Bt endotoxin gene and cow protease inhibitor
(CpTi) gene.
Transgenic plants have been produced by addition of one or more following
genes.
Bt endotoxin from Bacillus thuringiensis
Protease inhibitors
-Amylase inhibitors
Lectins
Enzymes
a. Bt endotoxin gene: The gram positive bacteria Bacillus thuringiensis
produces a crystal toxin called (delta) endotoxin.
a.
b.
c.
d.
e.

Crystal protein
Cry I
Cry II
Cry III
Cry IV
Cyt gene

:
:
:
:
:

Host
Lepidoptera
Lepidotpera and Diptera
Coleoptera
Diptera
Cytolytic
and
haemolytic
(Diptera)

There are two strains of B.thuriengiensis isolated and used against

insects. The strain, B.t. kurstaki is used against lepidoptan pests of forest
trees, vegetables, cotton and ornamentals and the another strain, B.t.
israelensis is used against ditperan insects
The endotoxin is a stomach poison and kills the lepidopteran insects if
consumed.
The gene (DNA fragment) responsible for producing endotoxin is
isolated from Bt and cloned into plants like cotton, potato, maize, etc. to
produce Transgenic cotton, etc.
Transgenic Bt plants

Target insect pests

Helicoverpa armigera, Spodoptera litura

1. Cotton

Pink bollworm
2. Maize
3. Rice

European corn borer

Leaf folder, stem borer

4. Tobacco, Tomato
5. Potato, Egg plant

Cut worms, Helicoverpa armigera

Colarado potato beetle

b. Protease inhibitors (PI) gene

Insects have proteases in their gut which are enzymes helping in
digestion of protein.

Protease inhibitors (trypsin-, chymotrypsin and

elastase like endoproteases) are substances inhibit the proteases and affect
digestion in insects. The protease inhibitor gene are isolated from one plant
and cloned into another to produce transgenic plants. In Plants, PI are
mainly restricted to storage organs (tubers, seeds etc.).
e.g. Transgenic apple, rice, tobacco containing PI.
Serine protease inhibitors
The first gene of plant origin to be transferred successfully to another
plant species resulting in enhanced insect resistance was isolated from
cowpea encoding a double headed trypsin inhibitor.

e.g.

Cowpea trypsin inhibitor (CpTI) is a PI isolated from cowpea and

cloned into tobacco.

This transgenic tobacco is resistant to Heliothis

virescens, Spodoptera litura and manduca sexta.

-Amylase inhibitor gene
-Amylase is a digestive enzyme present in insects for digestion of
carbohydrate. -Amylase inhibitor, affect digestion in insects.

Transgenic tobacco and tomato expressing -amylase inhibitor have

been produced which are resistant to Lepidopteran pests.
Cysteine protease inhibitors.
Two types of protein in cereal grains that inhibit insect digestive
enzymes and may play roles in preventing insects. Recently, a cysteine
protease inhibitor, orzacystatin, from rice has been isolated that inhibits
nearly all the protelolytic activity in the rice weevil and flour beetle midgut.
Recently, a cc (corn cystatin) was introduced into protoplast of rice
and the cystatin activity of the transgenic rice plants showed that 50 per
cent of the midgut protease activity in Sitophilus zeamais was inhibited.
c. Lectins genes
Lectins are proteins that bind to carbohydrates. When insect feed on
lectins, it binds to chitin in peritrophic membrane of midgut and prevents
uptake of nutrients.
e.g.

Transgenic tobacco containing pea lectin gene is resistant to H.

virescens
e. Enzyme genes
Chitinase enzyme gene, and cholesterol oxidase gene have been cloned
into plants and these show insecticidal properties.

Expression of an insect chitinase in transgenic tobacco enhances

resistance to some lepidopterans. Transgenic potato plants with BCH (bean
chitinase) gene were found to reduce the fecundity of the potato aphid,
Aphis solani.
PYRAMIDING GENES
To increase the protective efficacy, spectrum of activity and durability of
resistance, packages of different genes will be introduced into crops. It has
been found that low levels of serine protease inhibitors enhanced the
insecticidal activity of Cry1A, Cry3 and Cry4 against their respective target
insects.
Engineering transgenic crops with more than one gene to get multimechanistic resistance is called pyramiding of genes. e.g.
1. The CpTi gene and pea lectin gene were cloned to produce a tranagenic
tobacco.
2. Transgenic potato which express lectin and bean chitinase have been
produced.
Potentials/Advantages of Biotechnology in IPM
1. Slow development of resistance against transgenic Bt, PI, lectins
2. All plant parts express toxin and so no need for insecticide spray
3. No need for continuous monitoring
4. No environmental pollution, safe to NE, non-target organism
5. No cost of application for equipments and labourer
6. The use transgenic plants in one locality may affect the insect population
dynamics in other areas.