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Cloning in Plants

Applications of Genetic Engineering

Presented by:
Saba Anwar: 15211506-033
Aimen Aslam: 15211506-049
Asfa Riaz: 15211506-44
Syeda Nayyab Sammer:15211506-051
Presented to:
Mam Iqra Iqbal
Plant Cloning

• Plant cloning is the act of producing identical genetical plants from an

original plant.
• Simply cloning is just to take the cutting/clipping of a plant and grow
it elsewhere on its own.
• After 1-3 weeks, the roots will form from the cutting, and a new life of
a clone begins.

• Plants have been producing multiple numbers of genetically identical

copies of themselves from billions of years ago.
• Since the 20th century scientists start cloning in plants
• In 1964 a full carrot was cloned by F.C. Steward.
• Ever since then more and more plants have been cloned for supply,
money and the environment.
Importance of Cloning in Plants

• Cloning is a quick easy and cost-saving method of making new

• Cloning is an efficient way to keep the best genes as the new plants
will inherit the same qualities and characteristics of the mother plants.
• when the cuttings of the plants are inherently mature, they can also
flower fast after rooting.
• Cloning does not really affect the mother plant's health.
Techniques of Cloning

• Grafting and budding

• Marcotting or Air Layering
• Stem Cuttings
Grafting and Budding

• Grafting and budding are methods of asexual plant propagation

• That join parts from two different plants so they will grow as one
• These techniques are use in fruit trees and Rose Industries.
Plants Can Be Grafted
• Dicotyledonous and coniferous plants can be grafted.
• Monocotyledonous plants such ac lilies, Bamboo, and Grasses can‘t be
Graft Compatibility

Not all plants can be grafted easily even they are dicots or conifers.
• Within a variety
• Between variety
• Between species but in the same genus
• Between families- impossible
• Birch can’t be grafted on elm
Grafting Terms
Joining two plant pieces to make one plant
Basal part of the graft (understock or rootstock)
Detached shoot from last year’s growth with dormant buds,
upper graft part.
 Formation of graft union
This is a single layer of cells between the wood and bark. It must be
lined up for a good graft union.
Scion Preparation

• The scion must be dormant or not as advanced as understock.

• Scion wood should be healthy having well developed buds.
• Avoid wood with flower buds it would not graft.
Storage of scion wood
• Put in plastic bag and place in refrigerator at 40-50 degrees
• They can also be buried 12-18 inches deep in shaded soil for
several weeks.
Grafting Techniques

• The most common grafting techniques are:

• Whip grafting
• Cleft grafting
• Approach grafting
• Bridge grafting

• Budding is a form of grafting but instead of scion a small piece of

stem tissue with a bud or buds is used.
• Budding is done on immature plants and use in fruit and ornamental
• Budding season is from March-April budding last season wood and
June when budding on current season wood.
Three main types of Budding
• T budding
• Patch budding
• Chip budding
Marcotting/air layering

• Principle
Restricting the flow or interruption of the downward movement
of organic materials (CHO, growth regulators and other growth factors)
synthesized from the leaves and growing shoot tips, accumulating those
materials above the wound thereby stimulating rooting.
• Air layering is applied to mature branches within the tree crown
• A ring of bark is removed to promote photosyntahates accumulation.
• Exposed cambium is then treated with auxin powder to promote
• It is then wrapped in black polythene containing damp compost, peat
or other rooting medium.
Steps to Clone Plants

• Preparing
• Tools needed
• Rooting Hormones
• Select
• Cutting

• Tools needed
• A healthy and vibrant plant
• Sterilized sharp scissors or knife
• Rooting powder/liquid/gel (optional)
• Plastic/humor dome to cover the cloner. (optional)
• Rockwool cubes/Soils/a cup of water/cloning machine (depending on
the method you use for cloning)
• Rooting Hormone
• By nature, plants can produce the rooting hormones themselves after a
short span of time (Auxins)
• The rooting hormones help stir the plant cell growth and establish
roots faster
• Some plants can be easily clone without providing Synthetic Auxins
like tomatoes, mint, basil, rosemary, peppers
• While some plants such as fruiting or single harvest crops cannot be
clone without applying these hormones
• Select a healthy well-established plant, which does not indicate any
sign of diseases for weeks.
• Identify a vigorous branch that comes off from the main stem.
• Which will be in "v" form, from which new branchs regrows after the

• As new cuttings are very sensitive to microorganisms, bacteria, etc., be

sure to clean and sterilize the scissors/knife /razor before doing.
• Take a cutting at a 45-degree angle close to the main stem as this helps
increase the surface area, making it easy for the roots to sprout.
• Then place your cutting into a glass of water immediately.
• Doing this helps prevent the oxygen exposure because the new cutting
is somewhat sensitive.
Cloning methods

• Rockwool method
• Potting soil method
• Water cloning technique
• Cloning machine technique
• Cloning by Tissue culture

• Soak the rockwool cubes in neutral water overnight (or several hours)
to bring their pH down.
• Dip cutting into a rooting hormone for 15 - 30 seconds.
• If the hormone is in the powder form, make sure the cutting' s end gets
a little wet before applying
• Place the stem into the rockwool
• Mist it daily to moist the growing environment.
• Place the cloner under normal daylight
Potting Soil Technique

• Easy yet efficient method.

• Get the soils saturated enough
• Dip your cutting into the rooting hormones for 15 - 30 seconds,
• then place it into the potting soil
Water Cloning Technique
• First, fill the cup/plastic bottle with 3/4 of tap/distilled
• water of the room temperature should be get in the cup
• Check to ensure the pH level of your water is at between
5.5 to 6.0.
• Cut a piece of cling-wrap or plastic to cover and wrap
over the top of the cup
• Use a tip of a pen, or scissors to poke a hole in the cling-
wrap or plastic.
• Make sure the hole size is smaller than the cutting stem
to keep it tight into the pot.
• Now, put the cutting into the pot, keeping its end at least
5 cm under the water.
• Place the cloning pot under the indirect sunlight, or low
grow lights.
Cloning Machine Technique
• Efficient and quickest way to propagate plants.
• Cutting is not immersed in water but is constantly misted with low-pressure water, which
avoids diseases for the fragile roots
• There's plenty of oxygen for the roots
• you can do multiple clones at one time.
• Method
• Set up the cloning machine as per the manual.
• Fill with the water at the indicated level.
• Again, dip your cutting into the rooting hormones, then placed it into the neotypes.
• After that, run the cloning machines
Cloning by Tissue culture

• It works with small pieces of plants, called explants.

• These are grown in vitro using sterile agar jelly that contains plant
hormones and nutrients.
• This is an important way to preserve rare plant species or grow
commercially in larger nurseries.
Uses of Plant Cloning in the Modern World

• Producing more plants for the environment keeping it more safe and
healthy, this in turn helping people have healthier lives.
• Producing more plants for general use (decorations, in houses, garden,
pretty much everywhere)
• Growing a larger food supply for society and this can also benefit
countries or areas affected with a low supply of food.
• Scientific research with genetics and cell structure
Advantages of Plant Cloning

• Duplication can lead to more plants in the world

• A continuous supply of plants forever (some cloned plants are still alive from hundreds of
years ago
• The cloning procedure (grafting) can make genetically different root and shoot systems to
make a plant more better, stronger, longer lasting and have better qualities than the
• Plants can be made to not be infected by diseases and viruses
• The Plant Cloning Process can be made faster
• The Plant Cloning Process is sometimes less expensive than crop growing
• The pollination of plants may not be needed on cloned plants
• Some seeds are difficult to obtain or germinate so cloning is a preferable alternative.
• Much more profit to the food industry and the plant industry
Disadvantages of Plant Cloning

• Tends to eliminate diversity; more exposure to natural phenomenons

like disease. If a single disease harms one plant it will likely harm all
the crop, because they are all identical
• Cloning crops alters genes therefore disadvantaging crops by:
- causing higher mutation rates
- lower genetic variation (therefore less chance of survival)
- they have equal susceptibility to diseases
- can overtake other similar species and lead

A cloned plant affected by a

virus/ disease
Natural vs Cloned

• Natural Plants Cloned Plants

• Ever since the beginning of time Billions of years ago plants began to clone themselves. A
plants have been reproducing parent plant had a part of it broken off and grown into a new
and creating new plants with genetically identical plant. Now nearly 1 out of the 4 plants
different genetic makeups and you see every day is a cloned plant from a tree to a flower.
characteristics. When humans create cloned plants it is a whole different
thing. Now cloned plants can:
• They have evolved over time  have longer life spans
and have changed in appearance  have better appearances
and quality. They have grown  be stronger and able to withstand various things
naturally, have a normal  be invulnerable to diseases and viruses
lifespan, experience diseases and  have a larger supply
sicknesses, all need water and
sunlight and most need care
from people.
Applications of Genetic Engineering
Applications in Plants
Why genetic engineer plants?
• To improve agricultural, horticultural and ornamental value of crop
• Serve as bioreactor for the production of economically important
• For studying the gene action
General mechanism
Applications of modified plants
• To improve nutritional quality
• Insect resistance
• Herbiside resistance
• Delay fruit ripening
Improved nutritional value
• Increse linec acid and stearic acid
• Gene from E.coli
Delay fruit ripening
• Increase shelf life
• Decrease ethylene production
Insect resistance
• Pestiside resistance
• Soil bacteria:
• Gene from Bascillus thuringiensis
Subgenic plants
• Chines scientist creat strains of whe at that is resistant to powdery
• The strain lacks genes that encode proteins that repress defense
against mildew.
Genetic engineering in medicine
Insuline production
• Insulin gene from human DNA is removed and culture in bacteria to
produce human insulin.
Gene therapy
Severe Combined immune deficiency
• Immune disorder in children
• Gene codes for the enzyme adenosine deaminase
• Caused by mutation in gene on chromosome
• Abnormal ADA do not bind to deoxyadinosine
• Cause increase of deoxyadinosine level
• Kill B and T cells of the immune system
• Cure by gene therapy:
• The therapeutic gene called ADA in, to the bone marrow cells of
patient in laboratory followed by transplantation to patient
Chronic Granulomatus Disorder (CGD)
• Genetic disease in immune system
• Unable to produce immune protein
• Normal genes are introduce through bone marrow cells
• Then normal cells are able to produce
Inheritid blindness
• Cure blindness of inherited condition
• Caused by abnormality In gene called RPE65
GM Crops
• Since their introduction in 1996, genetically modified (GM) plants
have become an important component of our food supply.
• One genetic modification uses bacterial genes that produce a protein
known as Bt toxin.
• This toxin is harmless to humans and most other animals, but
enzymes in the digestive systems of insects convert Bt to a form that
kills the insects.
• Plants with the Bt gene do not have to be sprayed with pesticides.
• In addition, they produce higher yields of crops.
GM Animals
• Transgenic animals are becoming more important to our food supply.
• About 30 percent of the milk in U.S. markets comes from cows that have
been injected with hormones made by recombinant-DNA techniques to
increase milk production.
• Pigs can be genetically modified to produce more lean meat or high levels
of healthy omega-3 acids.
• Using growth-hormone genes, scientists have developed transgenic salmon
that grow much more quickly than wild salmon.
• Scientists are working to combine a gene for lysozyme—an antibacterial
protein found in human tears and breast milk—into the DNA of goats.
• Milk from these goats may help prevent infections in young children who
drink it
Application in Industries:
• Almost everything we eat and much of what we wear come from
living organisms
• Researchers have used genetic engineering to try to improve the
products we get from plants and animals
• Genetic modification could lead to better, less expensive, and more
nutritious food as well as less harmful manufacturing processes.
Oil Industry
• Plants store oil in their seeds (e.g., ground nut, mustard, rapeseed,
sunflower, sesamum, soya bean etc.) or in fruits (e.g., olive, avocado,
oil palm etc.)
• Such vegetable oils are used either as food or in industrial purposes
• According to various requirements the fatty acid quality and yield can
be improved by using genetic engineering technology
For example
• he wild type canola (rapeseed) contains 60% oleic acid along with
some other fatty acids
• A genetically engineered canola was constructed with a single gene
from the California bay tree that encodes an enzyme involved in the
synthesis of lauric acid
• These transgenic canola seeds contain 60% lauric acid instead of 60%
oleic acid; the new oil resembles the oil found in coconut and palm oil
• Similarly, a genetically engineered soya bean with desirable
monounsaturated fatty acid content of about 25% oleic acid to over
85% in its oil has been developed
• Such improved variety of soya bean oil is very good for health. The
production of such genetically engineered oil has already been