Transgenic plants

Introduction
• Transgenic plants are plants that have been genetic
ally engineered, a breeding approach that uses reco
mbinant DNA techniques to create plants with new
characteristics.They are identified as a class of gene
tically modified organism (GMO).
• Transgene?
• The genetically engineered gene added to the speci
es.
Advancements in Transgenic
plants
• Flavr savr tomato
• Golden rice
• Bt cotton
• Herbicide resistance
Flavr-savr tomato
• First genetically engineered crop granted license for
human consumption
• Produced by Californian company Calgene 1992.Cal
gene introduced a gene in plant which synthesize a
complementary mRNA to PG gene and inhibiting th
e synthesis of PG enzyme.
• On May 21, 1994, the genetically engineered Flavr S
avr tomato was introduced
 Genes involved
• IN CASE OF TOMATOES:
• i) pTOM5 encodes for phytoene synthase which pro
mote lycopene synthesis that gives red coloration.
• ii) p TOM6 gene encodes for polygalacturonase. This
enzyme degrades the cell wall, resulting in fruit softe
ning.
• iii) pTOM gene encodes for ACC oxidase. This enzyme
catalyzes the ethylene formation that triggers the frui
t ripening.
Procedure
• Softening of fruits is largely due to degradation of cell wall
(pectin) by enzyme polygalacturonase (PG). The gene encod
ing PG has been isolated and cloned (pTOM6)
• Procedure involves:
• 1) Isolation of DNA from tomato plant that encodes the enz
yme polygalacturonase (PG).
• 2) Transfer of PG gene to a vector bacteria and production o
f complementary DNA (cDNA) molecules.
• 3) Introduction of cDNA into a fresh tomato plant to produc
e transgenic plant.
Golden rice
• Golden Rice technology is based on the simple prin
ciple that rice plants possess the whole machinery t
o synthesis of B-carotene, and while this machinery
is fully active in leaves, parts of it are turned off in t
he grain endosperm.
• By adding only two genes, a plant phytoene synthas
e (psy) and a bacterial phytoene desaturase (crt I), t
he pathway is turned back on and B-carotene conse
quently accumulates in the grain.
Genes involved
• Golden rice was created by transforming rice with o
nly two B-carotene biosynthesis genes:
• 1). psy (Phytoene synthase) from daffodil (Narcissus
pseudonarcissus)
• 2). crt (Carotene desaturase) from the soil bacteriu
m Erwinia uredovora
Advantages
• Gene added that produces beta carotene (a red/ora
nge important for maintaining health)this gives rice
the golden colour.
• Beta carotene is needed to make vitamin A, which h
elps with eyesight.
• Can be used in areas where vitamin A deficiency is c
ommon, so it helps to prevent blindness.
Bt cotton
• Bt is a family of proteins originating from strains of t
he bacterium Bacillus thuringiensis.
• There are more than 200 different types of Bt toxins
, each affecting different types of insects. The Bt co
tton plants are genetically modified by the addition
of genes encoding toxin crystals in the Cry group of
endotoxin.
Mechanism of action
• The Bt cotton plants are genetically modified by the
addition of genes encoding toxin crystals in the Cry
group of endotoxin. When ingested by insects, the
Cry toxins are dissolved and activated by the high p
H environment of the animal's gastrointestinal syste
m.
Gene involved
• The cotton incorporated the cry1Ac gene from the s
oil bacterium Bacillus thuringiensis (Bt), making the
cotton toxic to bollworms. ... Monsanto has admitte
d that the pink bollworm is resistant to first genera
tion transgenic Bt cotton that expresses the single B
t gene (Cry1Ac).
How Bt works?
• 1. Ingestion
• 2. Solublization & proteolytic activation
• 3. Binding to target site
• 4. Formation of toxic lesions
Herbicide resistance
• How do these herbicides work?
• These herbicides target key enzymes in the plant m
etabolic pathway, which disrupt plant food producti
on and eventually kill it.
• So how do plants elicit tolerance to herbicides? So
me may have acquired the trait through selection o
r mutation; or more recently, plants may be modifie
d through genetic engineering.
Approaches
• A) modification of plant enzyme or other sensitive b
iochemical target of herbicide action to render it ins
ensitive to the herbicide, or inducing over producti
on of the unmodified target protein permitting nor
mal metabolism to occur.
• B) introduction of an enzyme or enzyme system tha
t detoxifies the herbicide in the plant before it reac
hes thesite of action.
Glyphosate-tolerant crops
• Glyphosate herbicide kills plants by blocking the EP
SPS enzyme, an enzyme involved in the biosynthesi
s of aromatic amino acids, vitamins and many secon
dary plant metabolites. There are several ways by w
hich crops can be modified to be glyphosate-toleran
t. One strategy is to incorporate a soil bacterium ge
ne that produces a glyphosate tolerant form of EPS
PS. Another way is to incorporate a different soil ba
cterium gene that produces a glyphosate degrading
enzyme.
Glufosinate-tolerant crops

• Glufosinate herbicides contain the active ingredient

phosphinothricin, which kills plants by blocking the
enzyme responsible for nitrogen metabolism and fo
r detoxifying ammonia, a by-product of plant metab
olism.
• Crops modified to tolerate glufosinate contain a bac
terial gene that produces an enzyme that detoxifies
phosphinothricin and prevents it from doing damag
e.
Advantages
• Excellent weed control and hence higher crop yields.
• Flexibility – possible to control weeds later in the pla
nt’s growth.
• Reduced numbers of sprays in a seasonm
• Reduced fuel use (because of less spraying).
• Reduced soil compaction (because of less need to go
on the land to spray).
• Use of low toxicity compounds which do not remain
active in the soil.