INVESTIGATORY

PROJECT IN
BIOLOGY

Genetic Modification and

Genetically Modified
Crops

PREPARED BY:

Muzammil M
CLASS: XII
SECTION: BB1
ROLL NO.: 12014
 Certificate

This is to certify that MUZAMMIL M of Class XII Sec.-

BB1 has satisfactorily completed the project work in
Biology prescribed by the Central Board of Secondary
Education (CBSE) for the year 2024 - 2025.

Signature of the teacher:

PAGE 2
Acknowledgement
 Content
S.No Topic Page
No.
01. INTRODUCTION
01
02. GENETICALLY MODIFIED ORGANISM
02
03. PROCESS OF GENETIC MODIFICATION
03
04. EXPERIMENT
04
07. GOLDEN RICE – 1 & 2
11
08. ARCTIC APPLE
14
09. FLAVR SAVR TOMATOES
16
10. INNATE POTATO
18
11 CONCLUSION
20
13 BIBLIOGRAPHY
21

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Introduction
WHAT IS A GENE?
A Gene is a fundamental, physical and functional unit of
heredity. It is responsible for the physical and inheritable
characteristics of an organism.
Genetic Engineering
It is manipulation/alteration of structure of a gene to
create a desired characteristic in an organism. If genetic
material from another species is added to the host, the
resulting organism is called transgenic.

Humans have altered the genomes of species for

thousands of years through artificial selection and more
recently mutagenesis.
 GENETICALLY MODIFIED
ORGANISMS

➢ Genetically modified organisms (GMO)

are organism whose genetic material has
been altered using genetic engineering
techniques.
➢ These techniques, generally known as
recombinant DNA technology, use DNA
molecules from different sources, which
are combined into one molecule to create
a new set of genes.
➢ This DNA is then transferred into an
organism, giving it modified or novel
genes.
➢ Transgenic organisms, a subset of
GMOs, are organisms which have
inserted DNA that originated in a
different species.
 Process Of Genetic
Modification

➢ All genetic changes affect the

protein synthesis of the organism.
➢ By changing which proteins are
produced, genetic engineers can affect
the overall traits of the organism.
➢ Genetic modification can be
completed by a number of different
methods as below:
1. Inserting new genetic material randomly or
in targeted locations
2. Direct replacement of genes (recombination)
3. Removal of genes
4. Mutation of existing genes

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 Experiment on Genetic modification
of a crop with Agrobacterium
Aim :-
To observe and understand Genetic
Modification of crops using “Agrobacterium
Method”.
Principles :-
• Agrobacterium-mediated transformation is
the efficient transfer of T-DNA into host cells
by bacteria.
• This process involves the transfer of a
plasmid by the bacteria into plant cells
during infection, which integrates into the
nuclear genome to express its own genes and
affect hormonal balance.
• The bacteria also produce enzymes involved
in opines synthesis, which are used by the
bacteria as nutrients. The infection begins
with the entry of bacteria through wounded
sites, and the release of phenolic
acetosyringone (AS) enhances binding.
• The AS activates VirA proteins, which
activate VirG, binds to other vir genes, and
stimulates T-strand generation.
• Factors like VirD2 and VirB proteins also
bind to the T-strand, forming a T-complex.
The complex is passed into the nucleus by
nuclear target signals.

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 • The T-DNA strand is integrated into the
plant genome randomly as single or multiple
copies, usually through recombination.
• Although much is known about the
molecular biology of T-DNA transfer in
Agrobacterium cells, little is known about
the plant-encoded factors involved in the
process
Materials & Reagent Required :-
Materials
• Sterile 50 ml plastic tubes
• Autoclave

• Controlled Tissue Culture Rooms at 25°C with 16/8

hr light/dark period
• Shaker Incubator

• Vacuum pump

• Laminar hood for tissue culture

• Glassware (Beakers, cylinders, Petri dishes, Duran

bottles, and Flasks)

• Filter paper

• Parafilm

• Forceps and Scalpel

• Pipettes

• Centrifuge

• Spectrophotometer

• Tissue culture vessels

• Surgical blades

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Reagents
• Explant (Stems, embryo, cotyledons, or other tissues)
• Agrobacterium strain

• 13% Sodium hypochlorite

• B5 Medium

• Agar

• Tryptone

• Yeast Extract

• Sodium Chloride

• 35% Hydrochloric acid

• Sterile distilled water

• 75% Ethanol

• Sucrose

• Abscisic Acid

• Rifampicin

• Kanamycin monosulfate

• Gellan gun powder

• PCR primer star Mix

• Carbenicillin disodium salt

Medium Required for Gene Transfer

1. LB ( Lysogeny broth) medium for Agrobacterium
culture
2. Murashige and Skoog medium for seed germination
3. Cocultivation medium
4. Shooting Medium
5. Rooting Medium

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Procedure:-

a. Sterilization and Germination of seeds

• The seeds are sterilized with Cl2 for 1-2 hours, and the seeds are then
soaked in water for 2 hours at room temperature in a Petri dish.
• The seed coats are removed with forceps from the seed and are
further sterilized with 75% ethanol for 30 seconds. These are then
rinsed with 20 ml of the 3% sodium hypochlorite.
• The sterilized seeds are placed on Petri plates containing seed
germination medium and are incubated at 28°C for 2 days in the
dark. Each plate can contain about 15-25 seeds.

Agrobacterium-Mediated Gene Transfer (Transformation) in Plants.

b. Inoculum Preparation
• 2 ml of the LB medium containing rifampicin and kanamycin is
inoculated with a single Agrobacterium colony. The culture is then
incubated in the shaker incubator at 28°C.
• The Agrobacterium culture prepared is centrifuged at 4000g for 10
minutes. The supernatant is removed, and the pellets are cleaned
with a 1 ml MS liquid medium.
c. Preparation of the explants
• The seeds are pulled out of the germination medium and are placed
on empty sterile Petri plates with a stack of filter papers.
• The radicle is removed, and the seed is cut to remove ½ of the
cotyledons and endopleura.
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 • The cotyledons are separated with a sterile scalpel and are wiped off.
The detached cotyledons are collected into the MS liquid medium in
a sterile glass beaker.
• The MS liquid medium containing the cotyledons are poured into MS
liquid medium containing Agrobacterium and shaken gently.
• The glass beaker is covered with a container and sealed with film, and
ut into the desiccators attached to the vacuum pump.
• The explants are left on the medium for 5 minutes after two sessions
of vacuum infiltration.
• A sterile filter paper is packed on the cocultivation medium, and the
infected explants are placed on the filter paper with forceps. The
adaxial surface of the cotyledon is to be kept upwards.
• The Petri plates are then sealed and incubated in the dark for 2 days at
28°C.
d. Shoot Initiation
• The explants are then transferred to the shooting medium containing
kanamycin and carbenicillin in order to inhibit the growth
of Agrobacterium.
• Every plate can contain 5-6 explants that are then incubated under
light for 2-3 weeks at 25°C.
e. Regeneration
• Once the shoots begin to appear on the explants, these are pulled out
and placed on sterile Petri plates with a stack of filter paper.
• The shoots are cut off with a sterile scalpel, and the embryoid part of
the shoots are removed.
• The shoots are then transferred to a 100 ml glass flask or tissue culture
vessels with a rooting medium.
• About 3-4 shoots can be added per vessel. The vessels are incubated
under light for 1-2 weeks at 25°C.
• After 2 weeks, if no roots are observed, the unfolded leaves and end
parts of the shoots are cut off, and the shoots are transferred to a
new rooting medium.
f. Plant acclimation
• After the roots begin to appear, the over on the flasks are loosened
and further incubated at 25°C for 3 days.
• The plants are then pulled out from the medium and washed off with
running water.

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 • These are then transferred to pots filled with wet compost and
watered.
• The pots are covered with zip bags to keep the moisture. These are
incubated in a good condition chamber under light at 25°C for 1-2
weeks.
• Once the plants grow in good condition, the zip bags are removed,
and the plants are watered.

Limitations :-
• The technique of agrobacterium-mediated transformation
has limitations, including a narrow host range and limited
knowledge about plant-encoded factors affecting T-DNA
transfer efficiency.
• It is labor-intensive and requires detailed processes, often
prone to in vitro variations. Monocot transformation
success relies on embryos as explants, but these are only
available for a short period.
• Agrobacterium-mediated transformation cannot transfer
large DNA molecules into economically important plants,
suggesting the need for a powerful vector system.
• This highlights the need for more efficient and effective
methods.
Advantages of Genetic Modification in
Crops :-
➢ Modified to make them more resistant to unfavorable
conditions
➢ Produce higher yields
➢ Use less fertilizers
➢ Use lesser water
➢ Pest resistance

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 ➢ Herbicide tolerance
➢ Increasing food supplies in co-relation with an increasing
world population
➢ Insect Resistant
➢ Improved and Altered Nutrition
➢ Longer Shelf Life
➢ Faster growth

Results:-
The out product formed is disease resistive and the
rich in proteins and vitamins lacking in pure stage.

SOME IMPORTANT GENETICALLY

MODIFIED CROPS

1. GOLDEN RICE
2. ARCTIC APPLE
3. FLAVR SAVR TOMATO
4. INNATE POTATO

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Golden Rice

• Golden rice is a variety of rice (Oryza sativa) produced

through genetic engineering to biosynthesize beta-
carotene, a precursor of vitamin A, in the edible parts of
the rice.
• b-carotene is taken from Narcissus pseudonarcissus
(daffodil) and inserted into the genome of a temperate
strain of rice, using Agrobacterium tumefaciens (Crown
gall) as the vector.
• It is intended to produce a fortified food to be grown
and consumed in areas with a shortage of dietary
vitamin A.
• Golden rice was created by transforming rice with two
beta-carotene biosynthesis genes:
1. psy (phytoene synthase) from daffodil (Narcissus
pseudonarcissus)

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 2. crtI (phytoene desaturase) from the soil bacterium
Erwinia uredovora
• The psy and crt I genes were transferred into the rice
nuclear genome and placed under the control of an
endosperm-specific promoter, so that they are only
expressed in the endosperm.
• The exogenous lcy gene has a transit peptide sequence
attached, so it is targeted to the plastid, where
geranylgeranyl diphosphate is formed.
• The bacterial crt I gene was an important inclusion to
complete the pathway, since it can catalyse multiple
steps in the synthesis of carotenoids up to lycopene,
while these steps require more than one enzyme in
plants.
• The end product of the engineered pathway is lycopene,
but if the plant accumulated lycopene, the rice would be
red.
GOLDEN RICE – 2
• In 2005, a team of researchers at Syngenta produced
Golden Rice 2.

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• They combined the phytoene synthase gene from
maize with crtl gene from the original golden rice.
• Golden Rice 2 produces 23 times more carotenoids
than golden rice (up to 37 µg/g).
• This is because phytoene synthase gene of maize
is the most effective gene for carotenoid synthesis,
and preferentially accumulates beta-carotene (up
to 31 µg/g of the 37 µg/g of carotenoids).

PAGE 17
 Arctic apple (Waltz, 2015)

• Browning is caused by polyphenol oxidases (PPOs)

naturally present in fruit and vegetables.
• When fruit is cut or bruised, these enzymes
catalyze the oxidation of polyphenols to quinones,
causing oxidative browning.
• The damage is superficial but can affect the taste
and texture of the apple as well as its cosmetic
qualities.
• In the Arctic varieties, the GM apples were
genetically engineered with a transgene that
produces specific RNAs to silence the expression
of at least four browning PPO genes.
• The apple RNA sequences were introduced into
Granny Smith and Golden Delicious varieties, to

PAGE 18
 bind complementary RNA and form a double
strand.
• As RNA is single stranded, the double-stranded
sequence is read as a mistake, and the plant's
naturally occurring Dicer enzymes are sent to chop
it up, resulting in no or significantly fewer PPO
proteins being produced.

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 Flavr Savr Tomato

• It is a genetically modified tomato, was the first

commercially grown genetically engineered food to
be granted a license for human consumption.
• It was developed by the Californian company
Calgene in the 1980s.
• The tomato has an improved shelf-life, increased
fungal resistance and a slightly increased viscosity
compared to its non-modified counterpart.
• It was meant to be harvested ripe for increased
flavor for long-distance shipping.
• The Flavr Savr contains two genes added by
Calgene; a reversed antisense polygalacturonase
gene which inhibits the production of a rotting
enzyme and a gene responsible for the creation of
APH(3')II, which confers resistance to certain
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 aminoglycoside antibiotics including kanamycin
and neomycin.
• The Flavr Savr was made more resistant to rotting
by the addition of an antisense gene which
interferes with the production of the enzyme Beta
polygalacturonase.
• The enzyme normally contributes to spoilage by
degrading pectin in cell walls and results in the
softening of fruit which makes them more
susceptible to being damaged by fungal infections.

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Innate Potatoes
• It is designed to resist blackspot bruising,
browning and to contain less of the amino acid
asparagine that turns into acrylamide during the
frying of potatoes.

• Acrylamide is a probable human carcinogen, so

reduced levels of it in fried potato foods is
desirable.
• Though, browning does not affect the quality of
the potato, it is simply that consumers tend to
not want to purchase "damaged" or possibly
spoiled goods.
• The 'Innate' name comes from the fact that this
variety does not contain any genetic material
from other species (the genes used are "innate"
to potatoes) and uses RNA interference to switch
off genes.
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• Simplot hopes that not including genes from
other species will assuage consumer fears about
biotechnology.
• The "Innate" potato is not a single cultivar;
rather, it is a group of potato varieties that have
had the same genetic alterations applied using
the same process.
• Five different potato varieties have been
transformed, creating "innate" versions of the
varieties, with all of the original traits, plus the
engineered ones.
• Ranger Russet, Russet Burbank, and Atlantic
potatoes have all been transformed by Simplot,
as well as two proprietary varieties.
• Modifications of each variety involved two
transformations, one for each of the two new
traits.
• Thus there was a total of ten transformation
events in developing the different Innate
varieties.

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CONCLUSION

We have discussed the promising aspects of

Genetic Engineering that can bring about
tremendous changes in human life. However the
manipulation of living organisms by the human race
cannot go on any further without regulation. Some
ethical standards are required to evaluate the
morality of all human activities that might help or
harm living organisms. Going beyond the morality of
such issues the biological significance of such things
is also important. Genetic modification of organisms
can have unpredictable results when such organisms
are introduced into the ecosystem.

Every new technology aims to improve man’s

life. It is for man to make the judicious use of its
applications…

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Bibliography

➢www.encyclopedia.com

➢https://en.wikipedia.org/wiki/Genetics

➢https://unescobmw.org/

➢https://www.who.int/

➢https://fssai.gov.in/

➢https://www.ncbi.nlm.nih.gov/

➢Trueman’s Biology

➢NCERT Textbook

➢Concepts of Genetics (W. S. Klung)

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THANK
YOU

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