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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