Plant and Animal Biotechnology

Applications
Read Chapter 10
Humans dependent on crops for food
World population expanding
Requirement of:
Optimization of agriculture efficiency
Increase in productivity

How can this be achieved?
Increase crop yields (new varieties)
Increase nutrition value
Disease resistant plant/crops
Insect resistant plant/crops
Increase post harvest life


Agriculture (plants and animals)
Methods for improvement in quality and productivity of crops
1. Classical plant breeding

Practiced since ancient times

Breeds with different desired characteristics are used for deliberate interbreeding
(cross breeding)

Breed 1 (wheat) = Good yield but not resistant to disease

Breed 2 (wheat) = Poor yield but resistant to disease

Cross breed produces progeny (hybrid) with desired trait i.e. disease resistant +
high yielding crop (wheat)


Modern bio-techniques for improvement of
quality and productivity of crops
1. Micropropagation
2. Protoplast fusion
3. Genetic engineering of plants
Using agrobacter (dicot only)
Gene gun/particle gun/shotgun technique

Micropropagation (Clonal propagation)
The technique of asexual or vegetative propagation
(multiplication) of plants in vitro (test tubes) from small
sections of plant such as stem tip, root tip. node, meristem,
embryo or seed etc.
Steps/stages:
1. Selection of plant material (plant with desired trait)
2. Testing for no infection (virus/bacteria)
3. Establishment of aseptic culture (free from microbes)
4. Callus (mass of undifferentiated cells) formation
5. Multiplication (callus cells)
6. Shoot elongation (specific plant hormones)
7. Root formation (specific plant hormones)
8. Plant establishment in small pots
9. Transplantation in field


Micropropagation
Banana, Oil Palms, Coffee plants etc
The plus points

Rapid multiplication
Pathogen removal
Uniform plants (clones)
Germplasm stored, not seeds.
Embryos can be rescued.

Somaclonal variation
Cells from a single plant are taken and grown.
It can give rise to variations.
Normal Tissue culture
Reasons for variation:
Plant cells can be at different stage of growth
Growth conditions different
Environment conditions different
Mutations
Chromosome number variation

Protoplast Fusion
Potato, tomato, pepper, tobacco plants have been regenerated from protoplasts
Tomato
Potato
Hybrid
Pomato
Plant Genetic Engineering
First practical system for gene
transfer in plants in 1983 using
Agrobacterium tumifaciens
(Agrobactor)
Agrobactor contains Ti Plasmid
(T-DNA)
Tumor inducing (Ti) plasmid
Ti plasmid is transferred into
plant genome.
Ti Used as vector for inserting
gene in plants (foreign gene
incorporated in Ti plasmid)
Good for dicot plants because
agrobactor infects only dicots
Monocots: gene gun technique
Plant cells are bombarded with
plant DNA coated nanogold
particles.


Cotyledons: the first leaves to appear from a germinating seed
Why genetic engineering for plants?
Interspecies barriers to transfer genes
disappear.
Traditional breeding methods lack in precision.
Improved resistance to herbicides.
Improved resistance to plant pathogens and insect
pests.
Improved post harvest characteristics.

These are also application of plant genetic
engineering/protoplast fusion


Gene for Bt toxin incorporated into cotton, tomato and corn.
Bt gene produces a protein called Cry toxins in transgenic crops that kills pest and their larvae.
Therefore, makes it resistant to insect pests
Transgenic plant: genetically engineered plants
Insect resistant transgenic crops (Biological Control)
Example: Bt corn or Bt cotton
Bacillus
thuringiensis
Improving Post harvest
characteristics
NEW FLAVOR SAVER: A tomato modified to contain a flavor gene
from lemon basil won a taste test against regular tomatoes
Transgenic soybean plant
Healthier/high value cooking oils
Contains less saturated fat
Oil produced from these genetically
enginnered plant is good for heart patients
Ethical issues/Biosafety issues
How safe are these crops for human consumption?
Playing with god?
Toxic gene from transgenic plant (corn:fodder for cattle) can enter food crop by accident
Food produced from genetically engineered plants/animals: harmful? Allergies
Bioterrorist can misuse the technology/steal transgenic organisms for their use

Read chapter 13 and 14
Animal Biotechnology


Increase milk quantity and quality
Transgenic cow which produces milk with low
cholesterol for heart patients
Disease resistant animals/cattle
Increase size of animal/goat, pig, chicken for
meat production
Fast growing animal for meat production
Selective breeding aims to increase the frequency of a large number of genes
that work together with the remainder of the animals genome to produce the
desired phenotype (characteristic)

'Fat Cattle' - Duke of Bedford and The Triumphs of Selective Breeding
Transgenic Animal An animal that has acquired novel genetic material
by artificial means rather than by normal route of sexual reproduction.
OR genetically engineered animal.
First Transgenic animal Rat with growth hormone gene inserted.
How is transgenic animal
generated?
Foreign gene selection and construction (PCR+
vector).
Microinjection of DNA in to male pronucleus (cell
containing separate male and female
nucleus/not fused yet) of fertilized egg.
Implanting these eggs into surrogate mothers.
Developing the embryo to full term.
Checking stable and heritable incorporation of
foreign DNA in the offspring.
Demonstration of transgene being expressed
and producing the desired product.
Microinjection of DNA into pronucleus is the
main method of gene transfer. It is done by
fine glass needles that allow injection of
DNA into fertilized eggs.
Pronucleus
Transgenic animals are used for
biopharmaceutical (human proteins) manufacturing
Used as bioreactors
Production of human proteins of pharmaceutical value
in animal milk
Protein Animal Use
Antithrombin
III
Goat
Reduce the amount of blood needed in some
surgeries
Factor VIII,
Factor IX
Goat, Pig,
Sheep
Treatment of hemophilia (bleeding disorder)
(blood clotting protein)
CFTR Sheep Treatment of cystic fibrosis
Lactoferrin Cow Natural antibiotic and used in coronary surgery
Alpha-1-
antitrypsin
Sheep Treatment of cystic fibrosis and emphysema
Lysostaphin Cow
An anti-bacterial compound that prevents
mastitis in cows
Spider silk
protein
Goat
Production of ultra-strong, lightweight medical
and industrial materials
Biopharmaceutical
(biosynthetic human proteins used for therapeutic)
Insulin: is a hormone important for glucose metabolism in the body.
Insulin causes cells in the liver, muscle, and fat tissue to take up
glucose from the blood, storing it as glycogen in the liver and
muscle.
Insulin stops the use of fat as an energy source. When insulin is
absent (or low), glucose is not taken up by body cells, and blood
sugar rises causing diabetes
Somatostatin: growth hormone (used to treat dwarfism caused due
to deficiency of hormone in diseased child)
Interferons: glycoproteins made by our body to fight virus infection
and inhibit the growth of cancer cells. Used as medicine to fight
some virus infection/treat cancer.
Lymphokines: proteins made by immune system of our body to fight
various infections (bacteria/fungi/allergy etc)
Gene Therapy
Used for the treatment of diseases by the transfer of normal functional
gene in a patients body cells to restore the function of mutant abnormal
gene.
Two types:
1. Germ cell gene therapy
Healthy gene is introduced in the patients germ cells so that can be passed
on to the offsprings

2. Somatic cell gene therapy
Healthy gene is introduced in patients body cells (somatic) lacking it and is
not passed onto the offspring.

You should be able to answer the following:
(Important)
Define/short note: protplast fusion, gene gun, Ti
plasmid, Bt crops, micropropagation or clonal
propagation, Agrobacterium tumifaciens,
How are plants genetic engineered or transgenic
plants produced. (Ti plasmid used as vector)
How are transgenic animals made?
Application of transgenic animal/animal
biotechnology
Application of plant biotechnology