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BIOTECHNOLOGY

BIOTECHNOLOGY IN CROP IMPROVEMENT

• Biotechnology- Biology and Technology
• Controlled use of biological agents such as micro-
organisms or cellular components for beneficial use
• Development and utilization of biological processes,
forms and systems for maximum benefits to man and
other forms of life.

Biotechnology in India
• ICGEB – International Centre for Genetic Engineering
and Biotechnology in Triesta (Italy) and in New Delhi,
India (1987)
• In addition, the research centres for Lal Bahadur
Shastri Centre for Advanced Research in Biotechnology,
IARI, New Delhi
• National Dairy Research Institute, Karnal
• Indian Veterinary Research Institute, Izatnagar
• The Biotechnology centre of IARI is called National
Research Centre for Plant Biotechnology

Plant Biotechnology
Plant biotechnology aim at improving the genetic make
up, phenotypic performance or multiplication rates of
economic plants.

Two basic techniques used in plant biotechnology are

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i) Plant tissue culture
ii) Genetic engineering
Plant tissue culture
It is the cultivation of plant organs, tissues or cells in
vessels using artificial media and the techniques of plant cell
and tissue culture are also called in vitro technique.

History of plant tissue culture

Scientists Contribution
Schleiden and Cell theory - Cell is the functional
Schwann (1938 and unit of living organisms. Worked in
1939) Tradeschantia with knop’s solution.
Habertland (Father First attempt of plant tissue culture
of tissue culture)
(1902)
White (1934) First reported for the successful
continuous cultures of tomato root
tips in liquid culture and obtained
indefinite growth.
Morel and Martin Use of meristem tip culture to
(1952) obtain virus free Dahlias
Gautheret (1959) First hand book on plant tissue
culture
Murashige and Development of MS medium
Skoog (1962)
Guha and Development of haploids through
Maheshwari (1962) anther and pollen culture for the
first time.

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Power et al. (1970) First protoplast fusion
Larkin and Introduction of the term somaclonal
Scowcroft (1981) variation.
• Totipotency: The regeneration capacity or ability of a
plant cell to develop into a whole plant is known as
totipotency.

• Callus: A mass of unorganised regenerated cells in

culture medium is called callus
• Dedifferentiation: The conversion of mature cells into
the meristematic state leading to the formation of
callus
• Re-differentiation: The process of biochemical and
structural changes by which cells especially the
unorganised cells becoming specialized in form and
function with the ability to form a whole plant.
• Protoplast: Cell without cell wall (Naked cells)

Techniques of plant tissue culture

1. Explant
Plant tissue or organ excised and used for invitro
culture.

2. Surface sterilization
1. Explant surface should be sterilized to eliminate
bacteria , fungi
2. 1-2% solution of sodium or calcium hypchloriteor 0.1%
solution mercury chloride.

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3. Rinse the explant in sterilized distilled water to
remove disinfectant.
4. Under aspectic conditions- laminar flow chamber.

3. Sterilization
Microbes present in the culture media, culture vessels,
and instrument are inactivated by suitable treatment.
1. Flame: forceps, scalpel, needles-95% alcohol and
flamed.
2. Dry heat: test tube, culture flasks- heated on a
burner
3. Ethanol (70%): laminar air flow chamber, culture
vessels, hands of worker
4. Autoclaving: culture media, culture vessel at 1210C
and 15 psi for 15-20 min.
5. Airfilter: air blowing the laminar tools is sterilized
by HEPA filter.
6. Filter: thermobiable constituents like ABA, GA3,
enzymes 0.45µm pore size.

4. Nutrient medium/ culture medium

• Plant cell and organs are cultured
• Contain inorganic salts, vitamins, carbon sources
(sucrose)
• Growth regulators viz, auxin (Root formation) and
cytokinin (Shoot formation)
• Auxin: 2,4 D(0.5-2.0 mg/l), NAA & IAA (Natural),
Kinetin and Benzyl Amino Purine (BAP) commonly used
Kinetin Zeatin.

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• Organic supplements: coconut water, casein
hydrolysate and yeast extract
• PH of medium- 5.5 using1N KOH or HCL.
• Solidify agent - Agar (6 g/l): Commonly used media
(Murashige& Skoog).
• Callus culture: The cells on the agar medium develop
into an unorganized mass- callus.
• Suspension culture: In liquid medium, a suspension of
free cells and small cell masses. The medium is
autoclaved at 15psi for 15-20 min.

5. Environmental conditions
1. The organ and cell culture evaluated under controlled
temperature, light, RH.
2. Temperature : 18-250C
3. Light is not essential but beneficial for plantlet
regeneration
4. Culture room/ incubator.

6. Subculturing
After a period of time, it is necessary to transfer
organs and tissue to fresh media.
1. In tissue and cell culture portion of tissue is used to
inoculated in new culture tubes
2. Subculture of callus culture every 4-6 weeks
3. Suspension culture every3-14 days.

7. Plant regeneration and transfer to soil

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• Production of roots shoots from cell and tissue culture-
organ regeneration or organogenesis.
• Rice, maize, barley, oats, sugarcane, colocassia, potat,
pea, soyabean, chickpea, alfalfa.
• Transfer of whole plants from test tube to soil is easy.
• The plants may be pretreated prior to transfer to soil
to make them hardy.
• In laboratory, they have transferred in small pots and
cover with vessel eg. Inverted breakers- to present
excess transpiration.
• After 3-4 days covers are removed and kept in diffuse
light for 5-10 days.
• Hardening on a large scale mist chamber.
• The plants then transferred into greenhouse and after
1-2weeks planted in soil and kept in sunlight.

Classification of plant tissue culture techniques

Based on the plant part used as explants and type of
development in vitro
i) Embryo culture
ii) Meristem culture
iii) Anther or pollen culture
iv) Tissue and cell culture

1. Embryo culture (Hanning, 1904 - Cruciferace)

• Young embryos are excised from developing seeds and
are placed on a suitable nutrient medium to obtain
seedlings
• Two types of embryo culture

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1. Culture of immature embryos (hybrid seed) - eg:
Soybean, Barley.
2. Culture of mature embryos (Eg: Orchids, Iris).

Applications
1. Recovery of distant hybrids
• Prevention of embryo abortion in wide crosses by
using embryos from incompatible crosses.
• In intergeneric and interspecific hybridization, post
zygotic barriers prevent normal seed development
and cause embryo abortion.
Eg. H. vulgare x S. cereale; H. vulgare x Triticum spp.
• Tetraploid and hexaploid wheat carry two dominant
genes kr1 and kr2, prevent seed development with
secale.

2. Recovery of haploid plants from interspecific crosses

• H. vulgare x H. bulbosum – zygote formed but during
embryo development H. bulbosum is eliminated
and haploid embryos of H. vulgare formed – Bulbosum
technique.

3. Propagation of orchids
• Orchid seeds are naked – lack stored food.
• Embryo development is incomplete at the time the
seeds mature
• Young/mature embryos removed and develop into
seedling either directly or through callus formation.
4. Shortening the breeding cycle

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Embryos develop directly into seedlings without
undergoing formation of seeds. Thus the next
generation may be grown one or two weeks early than
seeds.
5. Overcoming dormancy
Eg. Iris, Prunus, Tarus.

2. Meristem culture (5-10 mm shoot apical with leaf

primordia)
• Cultivation of auxillary or apical meristems, particularly
of shoot apical meristem and regeneration in a suitable
culture medium.
• Involves pre-existing apical meristem and regeneration
of adventitious roots.
• Widely used for quick vegetative propagation of large
number of plants.

Applications
• It is used for micro-propagation (mass production of
clonal progeny through tissue culture) in banana
(commercially successful), Strawberries, Citrus and
some timber trees Dalbergies sissoo (Sisam).
• Production of virus free plants-eg: Potato, Sugarcane,
Tapioca.
• Exchange of germplasm of plantlets is safe.
• Meristems are suitable for cryopreservation (-196◦C
for long term storage).

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3. Anther culture/pollen /microspore culture (Guha and
Maheswari, 1964 in Datura)
• Haploid plants may be obtained from pollen grains by
placing anthers or isolated pollen grains on a suitable
culture medium.
• For successful anther, plants should be grown under
controlled temperature, light and humidity.
• In Tobacco, exposure of excised flower buds at 5oC for
72 hours prior to removal of anther enhances recovery
of haploid plants.
• Brief exposure of anthers to high temperature at 35 oC
for 24 hours in B. campestries.
• Media requirements vary with species Eg; Pollen grains
of Datura and Tobacco produce embryo on agar medium
2-4 % sucrose, 3% for barley, 6 % wheat and Potato.
• Sucrose is essential for anther culture.
• Optimum stage of pollen Eg: Datura, Tobacco-just
before or after first pollen mitosis. Cereals-Early or mid
uninucleate stage i.e, before first Tomato and A
.thaliana-PMC’s are meiosis-I. Brassica-Trinucleate
pollen grain.
• Maintained by altering periods of light (12-18 hr; 5000-
10000 lux) @ 28 oC and darkness (12-6 hr) @ 22 oC .
• The embryo or callus become 3-5 cm long then they are
transferred to medium for good root development.
• Pollen grains of cultured anthers show remarkable
cytological changes during first 6-12 days - inductive
period.

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Applications
1. Obtaining haploid plants
2. Haploids are useful in cytological studies
3. Doubling the chromosome number of haploids -
homozygous diploids within a year as compared to
inbreeding
Eg. Rice- Xin Xiu, Shin shu
Wheat- Hua pei 1, Lung Hua 1

4. Tissue and cell cultures

• Application of plant tissue and cell culture in crop
improvement depends on techniques for regeneration of
whole plants from them.
• Both callus and suspension cultures of many species
regenerate whole plants.
• The Somatic embryos regenerate to form whole plants
– eg. Carrot, wheat, soybean, coffee
• In shoot regeneration, shoot buds differentiate from
the cell mass followed by differentiation of roots –
Tobacco, rice, wheat, barley, coffee

Shoot regeneration
• Shoot buds differentiate from cell culture.
• Shoot buds are unipolar i.e., have only plumule and have
vascular connections with the callus tissue
• High cytokinin to auxin ratio supports shoot
regeneration, while a high auxin to cytokinin ratio
promotes root regeneration

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• In alfalfa, two step process of shoot regeneration
occurs; first callus is produced on a medium having high
2,4-D to kinetin ratio, second shoot regeneration occurs
when the callus is transferred to a growth regulator-
free medium.
• Gibberlliins suppress shoot regeneration

Somatic embryogenesis
• Somatic embryos originate from single peripheral or
deep-seated cells of callus.
• They form group of cells and it divides and progresses
through globular, heart-shaped, torpedo-shaped and
cotyledonary stages similar to zygotic embryos.

Application of tissue and cell cultures

The various uses of cell and tissue culture are
i) Clonal propagation
ii) Somoclonal variation
iii) Mutant isolation
iv) Biochemical production
v) Biotransformation

i) Clonal propagation
• Tissue cultures – Quick vegetative propagation of plant
species.
• Virus and disease free plants produced
• Used for asexual propagating crops

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• In some species, oil palm and date palm, somatic
embryogenesis offers the only route for micro-
poropagation.
• Individual SEs/ shoot buds may be enclosed in beads of
a suitable matrix eg. Calcium alginate – Artificial seeds
• The beads contain all the nutrients needed for
production of plantlets and protect from microbial and
pest attacks

Difficulties
1. Occurrence of genetic variation among the regenerated
plants
2. Reduced by using young tissue cultures

ii) Somoclonal variation

• The plants regenerated from cell cultures show
heritable variation for both qualitative and quantitative
traits - Somoclonal variation
• Eg. Sugarcane, potato, tomato – Jointless pedicel mutant
useful for mechanical picking

iii) Biochemical production

• Callus and suspension cultures of any species accumulate
certain biochemicals known as secondary metabolites.
• Secondary metabolites are organic compounds that are
not directly involved in normal growth, development or
reproduction of an organism.

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• Humans use secondary metabolites as medicines,
flavourings and recreational drugs
• Shikonin, drug and precious natural dye for silk and
cosmetics – produced from cell cultures of lithospermum
erythrozhizon.
• Berberine (Coptis japonica) and taxol (Taxus sp.) are
also produced from cell cultures

(iv) Mutant isolation

• Fiji disease inn sugarcane line isolated from cell culture
and released as a new variety (Ono)
• Bacterial wilt resistant tomato line was also released
• Wild fire disease of Tobacco – Pseudomonas tabaci.
Tobacco cells resistant to methionine sulfoximine –
similar to pathogenic toxin were isolated. Clones from
this resistant to wild fire disease.

Somatic hybridisation
• Usually sexual hybridization has been used for
transferring of various agriculturally important traits
into numerous cultivated crops.
• There is limitation to sexual hybridization, if it is
between cultivated species and their wild relatives on
account of many barriers.
• An alternative system for hybridization of plants,
"somatic cell hybridization" or otherwise known as
“parasexual hybridization”.
• Fusion of cells takes place through protoplasts
• Protoplasts are naked cells or cells without cell wall.

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• The somatic cells are manipulated by removing the cell
wall and protoplasts are fused.
• This process of hybridization at cell level is considered
as an alternative to sex.

Techniques of somatic hybridisation

It involves the following four steps
i) Isolation of protoplasts
ii) Fusion of protoplasts of the desired species/varieties
iii) Selection of somatic hybrid cells
iv) Culture and regeneration of hybrid plants

i) Isolation of protoplasts
Treating the cells/tissues with a suitable cellwall
degrading enzymes – mixture of pectinasse or macroenzyme
(0.1 – 1.0%) and cellulase (1-2%).

ii) Protoplast fusion

Two methods viz., Chemical and electrical
• In chemical method, Polyethylene Glycol, high PH (10.5)
and high Calcium ions (50mmol-1)
• In electrical method, electric current is passed to
induce fusion.
iii) Selection of hybrid cells
• After fusion, there is a mixture of parental types viz.,
homokaryons and heterokaryons
• Union of proptoplasts of same species – Homokaryons
• Union between protoplasts of two different species –
heterokaryons

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• Hybrid cells can be identified by
i) Hybrid vigour at the callus stage
ii) Morphology which is intermediate between two
parents
iii) Biochemical and molecular techniques such as
isozyme pattern and RFLP and PCR based markers.

iv) Culture and regeneration of hybrid cells

• Hybrid cells are cultured in a Murashige and Skoog (MS)
medium with suitable modification
• In culture medium, protoplasts regenerate a new cell
wall within 2-4 days and cell division starts within 2-7
days.
• ‘Somatic incompatability’ also occur in some somatic
combinations.
• Some somatic hybrid plants retain the full or nearly full
somatic complements of the two parental species –
symmetrical hybrids
Solanum tuberosum + Lycopersican esculentum
Arabidopsis thaliana + Brassica campestris
• Many somatic hybrids exhibit full somatic complement
of one species, while the other species lost during
mitotic divisions – asymmetric hybrids
Nicotiana tabacum + Daucus carota.

Cybrids
Cybrids or cytoplasmic hybrids are cells containing
nucleus of one species but cytoplasm from both the
parental species.

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Applications of cybrids
1. Transfer of plasmagenes of one species into the nuclear
background of another species in a single generation
2. Sexually incompatible combinations
3. Recovery of recombination between the parental
mitochondrial or chloroplast DNAs
4. Production of wide variety of combinations
5. They are used for the transfer of cytoplasmic male
sterility from Nicotiana tabacum to N. sylvestris.

CRYOPRESERVATION OF GERMPLASM/ FREEZE

PRESERVATION
The preservation of cells, tissues and organs in liquid
nitrogen (-196oC) is called cryopreservation and the science
dealing this activity is known as cryobiology.

Steps involved in cryo-preservation

The steps in cryo-preservation are as follows
i) Freezing
ii) Storage
iii) Thawing
iv) Reculture

i) Freezing
• Either slow (cooling rate 0.5 to 4 oC /min from 0oC to -
100 oC and then traasnfer to liquid nitrogen or rapid (>
1000 oC/min) or it may be initially slow 5 oC/min upto -30

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to -50oC held at this temperature for about 30 min and
then cooled rapidly into liquid nitrogen.
• Plant vitrification solutions like ethylene glycol, sucrose
are used for partial dehydration of the cells/tissues
before freezing.

ii) Storage
• Cryoprotectants - DMSO (5-8%), glycerol and proline
(10%) must be added to the culture medium
• It prevent the formation of large ice crystals in cells
and protect them from toxic solution effect due to
water loss from the cytoplasm
• Frozen cells and tissues stored in LN refrigerator the
temperature must not raise above -130oC, otherwise ice
crystals may be formed.

iii) Thawing
• Thawing of the frozen materials is achieved by plunging
vials into warm water (37-40oC for 90 seconds), washed
several times to remove the cryoprotectants and
preserved in ice bath.

iv) Re-culturing
• The cells/tissue/shoot tips may be transferred to the
required fresh medium after thawing for regeneration.
• In general, meristematic cells survive better during
cryopreservation than do mature differentiated cells.
The shoot tips are being used for germplasm storage of
at least some clonal crops like potato.

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Slow growth culture

• They are maintained either at low temperature (4-9oC)
or on a suitable medium haing high osmotic
concentration.
• High osmotic pressure can be maintained by adding 20%
sorbitol or high level of sucrose
• In addition nutrient status may be lowered down to
restrict the growth.
• Low oxygen and use of succinic acid, 2, 2, dimethyl
hydrazide, cycocel and abscicic acid are also used to
delay culture growth.
➢ The slow growth approach is being utilized for
germplasm conservation of specified root, tuber and
tree species by the NBPGR, New Delhi.
➢ National facility for Plant Tissue Culture Repository
has been created for this purpose and this developed
slow growth protocols for ginger, garlic, banana,
sweet potato.
➢ This approach is being used for conservation and
exchange of potato and sweet potato germplasm at
International Potato Centre, Lima, Peru.

Advantages of germplasm storage

1. Requires very small space
2. Free from diseases, pests and weeds
3. Materials can be stored for long periods
4. Errors in labelling minimized
5. Ideal for germplasm exchange

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6. Protection against natural disorders and preservation of
rare genomes

Molecular markers
• Those characters which can be easily identified are
referred to as marker characters.
• Markers are of three types
(i) Morphological markers – Shape, size, colour and
surface of various plant parts. Also known as
conventional markers
(ii) Biochemical markers – Variation in proteins and amino
acid banding patterns
(iii) Molecular markers – DNA fragements generated by
restriction endonuclease enzymes.
• There are two classes of DNA markers – i) PCR based
and ii) Non – PCR based
• Non - PCR based marker – RFLP
• PCR based marker – RAPD, AFLP, SSR, SNP, DNA
fingerprinting

RFLP – Restriction Fragement Length Polymorphism

RAPD – Random Amplified Polymorphic DNA
AFLP – Amplified Fragment Length Polymorphism
SSR – Simple Sequence Repeats
SNP – Single Nucleotide Polymorphism

Marker assisted selection

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• Indirect selection for a desired plant phenotype based
on the banding patterns of linked molecular markers.

Uses of molecular markers

• Construction of genome map
• Genetic mapping and tagging
• Germplasm characterization
• Marker aided selection

• The DNA fingerprinting was developed by Alec

Jeffreys (1985)
• Polymerase Chain Reaction technique, developed by
Kary Mullis (1985)
• Taq polymerase (Thermus acquaticus) is commonly used
DNA polymerse.

Characteristics of different molecular markers

Characteris RFLP RAPD AFLP SSR SNP
tic
Types of Single Multi Multi Single Single
visualisation locus locus locus locus locus
Allelism Biallelic Polyalleli Polyallel Bialleic Biallelic
c ic
Dominance Codomin Dominan Domina Codomin Codomin
ant t nt ant ant
Level of Good Good Excelle Excellen Excellen
polymorphis nt t t
m

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Polymorphis 2 to 5 Presence Presenc Multiple 2 alleles
m at the alleles / e/ alleles
locus level absence absence
Quantity of Large Small Small Small Small
DNA
needed
Quality of Good Good Good No Good
DNA restricti
needed ons
Reproducibil Good Low Good Good Good
ity
Time Long Fast Fast Fast Fast
Cost Expensi Average Averag Average Expensi
ve e ve
Technical High Medium Medium Low Medium
difficulty
Transgenic plants
• The most potent biotechnological approach for gene
transfer is the transfer of specifically constructed
gene assemblies through various genetic transformation
techniques – Genetic engineering.
• The plants obtained through genetic engineering
contain a foreign gene or genes usually from an
unrelated organism are called transgenes and the
plants containing transgenes are known as transgeneic
plants.

Steps involved in genetic engineering

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1. Isolation of gene of interest
2. DNA fragment preparation for the isolated interested
gene
3. Preparing a vector
4. Inserting the DNA fragment into the vector to
prepare the recombinant DNA
5. Introduction into host cell and stable maintenance
6. Selection of transformed cells
7. Recombinant DNA molecule is multiplies within the host
cell to produce a number of identical copies of the
cloned gene
8. Characterization and use of engineered organism

There are two major method involved in gene transfer

1. Agrobacterium mediated gene transfer
• It is a gram negative soil bacterium which infects
dicot plants.
• On infection of plant cells by Agrobacterium
tumifaciens the T DNA is integrated into the host
genome which causes crown gall disease.
• A recombinant Ti plasmid with a target gene
inserted into the T DNA can be integerated into the
plant DNA and made to be expressed.

2. Direct gene transfer

i) Chemical methods eg. PEG (Polyethylene Glycol)
ii) Electroporation
iii) Particle gun delivery
iv) Lipofection

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v) Microinjection and Macroinjection
vi) Pollen transformation
vii) Delivery via growing pollen tubes
viii) Laser mediated
ix) Fiber mediated

Among these methods, Agrobacterium mediated,

electroporation and particle gun delivery is the most
commonly used.

Applications of genetic engineering

1. Insect resistance
Bacillus thuringensis – Discovered by Ishiwaki (1901) in
diseased silkworms and named it as Bt by Ernst Berliner
from diseased flour moth larvae in thurinberg region of
Germany.
The cry gene of B.thuringensis produces a crystalline
protein called δ- endotoxin. The crystal proteins are
responsible for the insecticidal activities of the bacterial
strains.

The cry genes are grouped into different types

• Cry 1 (A-J) – insecticidal against lepidopteran pests
• Cry 2 A – Lepidopteran and Dipteran pests
• Cry 3 (A, B, C), Cry 7A, Cry 8A, 8B – Coleopteran pests
• Cry 4A, 4B, Cry 10A, 11A, 11B – Dipteran pests
• Cry 5A (A, b) – Nematodes
• Cry 5A (c), 5B – Hymenopteran pests

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2. Improvement in quality
• In tomato – ‘Flavr Savr’ helps in increasing the keeping
quality based on antisense RNA technology.
• In rice - ‘Golden rice’ rich in provitamin A by β carotene
synthesis in rice grain developed by Ingo potrykus and
Peter Bayer .
• Golden rice was created by rice with two beta-carotene
biosynthesis genes:
1. psy (phytoene synthase) from daffodil (Narcissus
pseudonarcissus)
2. crtI from the soil bacterium Erwinia uredovora
• Golden Rice 2, the phytoene synthetase gene (psy) from
maize and the carotene desaturase gene (crtl) from
Erwinia uredovora were inserted into rice.
• Golden Rice 1 contains about 1.6 g of total carotenoids
per gram of dry weight of grain. Golden Rice 2 contains as
much as 37 g total carotenoids per gram of dry weight of
grain.

3. Herbicide resistance
Roundup Ready soybean – Glyphosate herbicide
resistance

Examples of transgenic plants in some crop species

Crop Character modified Source of transgene
plants
Cotton Resistance to Bacillus thuringiensis
Helicoverpa
Herbicide resistant

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i) Glyphosate Microbial gene
ii) 2,4-D Acaligenes eutrophus
Maize Resistance to European Bacillus thuringiensis
Corn borer
Herbicide resistance to Arabidopsis thaliana
sulphonyl urea
Wheat Herbicide resistance to Streptomyces
glufosinate
Tobacco Insect resistance Vigna ungiculata
Brown spot resistance Serrati marcescens
Seedling blight Phascolous vulgaris
resistance
Freezing resistance Winter flounder fish
Cold resistance Arabidopsis thaliana
Herbicide resistance to Acaligenes utrophus
2,4-D
Potato Increased mannitol Escherichia coli
Resistance to potato Potato leaf roll
leaf roll
Resistance to x and y Potato virus x and y
viruses
Herbicide resistance to Streptomyces
glufosinate hygroscopicus
Increased starch Escherichia coli
content
Protein quality Seru albumin gene
from human
Tomato Insect resistance Bacillus thuringiensis

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Herbicide resistance to Streptomyces
glufosinate
Freezing resistance Fish (Winter
flounder)
Resistance to tomato Tomato mosaic virus
mosaic virus
Rapeseed Keeping quality Lycopersican
esculentum
Herbicide resistance
i) Sulphonyl Urea Arabidopsis thaliana
ii) Glufosinate Streptomyces
Male sterility Bacillus
amyloliquefaciens
Soybean Herbicide resistance to Microbial gene
glyphosate
Linseed Herbicide resistance to Microbial gene
i)Glyphosate Arabidopsis thaliana
ii) Sulphonyl urea
Alfalfa Protein quality Chicken
Transgenic facts
• First Genetic Engineering company is Genetech (1976)
• First transgenic plant developed in Tobacco (Fraley et
al. 1984)
• First transgenic variety cultivated in USA was ‘Flavor
savr’ of Tomato for delayed ripening (1994)
• First transgenic crop approved by GEAC for release in
India (Cotton – MECH 12, 162 & 184)

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• Highest area covered by transgene in the world is
soybean
• Highest area covered by transgene in India is cotton
• Larger area transgenic crop is cultivated in USA
• Maximum transgenics available in rapeseed and mustard
• Maximum transgenics is developed for herbicide
resistance trait
• India occupies 6th position in area

AGRIQUIZ.2K19
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