BT 101P.roy Plant&Animal Biotech

8/12/2019 BT 101P.roy Plant&Animal Biotech

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Plant and Animal Biotechnology

Applications


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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/cropsInsect resistant plant/crops

Increase post harvest life

Agriculture (plants and animals)


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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)


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Classical Plant breeding


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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


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Micropropagation (Clonal propagation)

The technique of asexual or vegetative propagation

(multiplication) of plants in vi tro(test tubes) from smallsections 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 pots9. Transplantation in field

Callus


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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.


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Somaclonal variationCells 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


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Protoplast Fusion

Potato, tomato, pepper, tobacco plants have been regenerated from protoplasts

Tomato PotatoHybrid

Pomato


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Plant Genetic Engineering First practical system for gene

transfer in plants in 1983 usingAg robacter ium tum ifaciens

(Agrobactor) Agrobactor containsTi Plasmid

(T-DNA)

Tumor inducing (Ti) plasmid

Ti plasmidis transferred intoplant genome.

Ti Used as vector for insertinggene in plants (foreign geneincorporated in Ti plasmid)

Good for dicot plants becauseagrobactor infects only dicots

Monocots: gene gun technique

Plant cells are bombarded withplant DNA coated nanogoldparticles.


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Plant genetic engineering


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Why genetic engineering for plants?

Interspecies barriers to transfer genesdisappear.

Traditional breeding methods lack in precision. Improved resistance to herbicides.

Improved resistance to plant pathogens and insectpests.

Improved post harvest characteristics.

These are also application of plant geneticengineering/protoplast fusion


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Gene for Bt toxin incorporated into cotton, tomato and corn.

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


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Improving Post harvest

characteristics


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NEW FLAVOR SAVER:A tomato modified to contain a flavor genefrom lemon basil won a taste test against regular tomatoes


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Transgenic soybean plant

Healthier/high value cooking oils

Contains less saturated fat

Oil produced from these geneticallyenginnered plant is good for heart patients


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Ethical issues/Biosafety issuesHow 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

Bioterrorist can misuse the technology/steal transgenic organisms for their useFood produced from genetically engineered plants/animals: harmful? allergies

Read chapter 13, 14 and 15, Biotechnology, J. E. Smith, Fifth edition


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Animal Biotechnology

Increase milk quantity and quality

Transgenic cow which produces milk with low

cholesterol for heart patientsDisease resistant animals/cattle

Increase size of animal/goat, pig, chicken for

meat production

Fast growing animal for meat production

Wool quality and quantity

Production of low cost pharmaceuticals and

biologicals


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Selective breedingaims 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


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Transgenic AnimalAn animal that has acquired novel genetic material

by artificial means rather than by normal route of sexual reproduction.

OR genetically engineered animal.

First Transgenic animalRat with growth hormone gene inserted.


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How is transgenic animal

generated? Foreign gene selection and construction (PCR+

vector).

Microinjectionof DNA in to pronucleus(cellcontaining 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.


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Zygote

Pronucleii


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Microinjection of DNA into pronucleusis the

main method of gene transfer. It is done by

fine glass needlesthat allow injection of

DNA into fertilized eggs.

Pronucleus

T i i l d f


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Transgenic animals are used for

biopharmaceutical (human proteins) manufacturing

Used as bioreactors

Production of human proteins of pharmaceutical value

in animal milk

The large majority of biopharmaceutical products are

pharmaceuticals that are derived from life forms.


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Protein Animal Use

Antithrombin

IIIGoat

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 CowAn anti-bacterial compound that prevents

mastitis in cows

Spider silk

proteinGoat

Production of ultra-strong, lightweight medical

and industrial materials

Some examples of animals as biopharmaceuticals


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Animal cloning (making same copy of

an animal)


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Potent applications of somatic

cell nuclear transfer Production of genetically identical laboratory

animals

In animal breeding, producing multiple copies of

supreme animals that could be utilized foranimal farming

Providing more reliable ways of producing

transgenic animals with less wastage

Genetic conservation; storing frozen semen and

embryos is expensive.


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Biopharmaceutical(biosynthetic human proteins used for therapeutic )

Insulin: is a hormoneimportant for glucose metabolism in the body.Insulin causes cells in the liver, muscle, and fat tissue to take upglucose from the blood, storing it as glycogen in the liver andmuscle.

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 bloodsugar rises causing diabetes

Somatostatin: growth hormone (used to treat dwarfism caused dueto deficiency of hormone in diseased child.)

Interferons:glycoproteins made by our body to fight virus infectionand inhibit the growth of cancer cells. Used as medicine to fightsome virus infection/treat cancer.

Lymphokines:proteins made by immune system of our body to fightvarious infections (bacteria/fungi/allergy etc)

G Th


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Gene TherapyUsed 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 (sperm and egg) so that

can be passed on to the offsprings

2. Somatic cell gene therapyHealthy gene is introduced in patients body cells (somatic) lacking it and is

not passed onto the offspring.


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You should be able to answer the following:

(Important)

Define/short note: protplast fusion, gene gun, Tiplasmid, 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


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Read chapter 8 and 9,

Biotechnology, J. E. Smith,

Fifth edition

BT 101P.roy Plant&Animal Biotech

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