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Plant and Animal Biotechnology Applications

Agriculture (plants and animals) 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

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)

Classical Plant breeding

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

Callus

Micropropagation
The plus points

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

Banana, Oil Palms, Coffee plants etc

Normal Tissue culture

Somaclonal variation
Cells from a single plant are taken and grown. It can give rise to variations.

Reasons for variation: Plant cells can be at different stage of growth Growth conditions different Environment conditions different Mutations Chromosome number variation

Protoplast Fusion
Tomato Hybrid Pomato Potato

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

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.

Plant Genetic Engineering

Plant genetic engineering

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

Transgenic plant: genetically engineered plants Insect resistant transgenic crops (Biological Control) Example: Bt corn or Bt cotton
Bacillus thuringiensis

Gene for Bt toxin incorporated into cotton, tomato and corn. Makes it resistant to insect pests

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

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

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 Wool quality and quantity Production of low cost pharmaceuticals and biologicals

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 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.

Pronucleii

Zygote

Pronucleus

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.

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.

Some examples of animals as biopharmaceuticals


Protein Antithrombin III Factor VIII, Factor IX CFTR Lactoferrin Animal Use Reduce the amount of blood needed in some surgeries Treatment of hemophilia (bleeding disorder) (blood clotting protein) Treatment of cystic fibrosis Natural antibiotic and used in coronary surgery

Goat Goat, Pig, Sheep Sheep Cow

Alpha-1antitrypsin
Lysostaphin Spider silk protein

Sheep
Cow Goat

Treatment of cystic fibrosis and emphysema


An anti-bacterial compound that prevents mastitis in cows Production of ultra-strong, lightweight medical and industrial materials

Animal cloning (making same copy of an animal)

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 for animal farming Providing more reliable ways of producing transgenic animals with less wastage Genetic conservation; storing frozen semen and embryos is expensive.

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 (sperm and egg) 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

Read chapter 8 and 9, Biotechnology, J. E. Smith, Fifth edition

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