Hrre is the brif information about plant tissue culture

1. PLANT TISSUE
CULTURE
Prepared by:
Mr. Abhay Shripad Joshi
Assistant Professor
Yash Institute of Pharmacy, Aurangabad
abhay.joshirss@gmail.com

2. PLANT TISSUE
CULTURE

3. PLANT TISSUE CULTURE
• Plant tissue culture is a collection of
techniques used to maintain or grow plant
cells, tissues or organs under sterile
conditions on a nutrient culture medium
of known composition. Plant tissue
culture is widely used to produce clones
of a plant in a method known
as micropropagation .

4. Advantages of Plant Tissue Culture
• The production of exact copies of plants that
produce particularly good flowers, fruits.
• To quickly produce mature plants.
• The production of multiples of plants in the
absence of seeds.
• The regeneration of whole plants from plant
cells that have been genetically modified.

5. • The production of plants in sterile containers that
allows them to be moved with greatly reduced
chances of transmitting diseases, pests, and
pathogens.
• The production of plants from seeds that
otherwise have very low chances of germinating
and growing, i.e.: orchids and Nepenthes.
• To clear particular plants of viral and other
infections and to quickly multiply these plants as
'cleaned stock' for horticulture and agriculture.

6. ADVANTAGES OF PLANT TISSUE
CULTURE
1) Availability of raw material
2) Variation in supplies and quality
3) Patent rights
4) Easy purification of the compounds
5) Modification of chemical structure
6) Disease free and desired product
7) Crop improvement

7. EXPLANT
• Any part of a plant taken out and grown
in test tube under sterile conditions in
special nutrient media is called explant.

8. METHODS OF PLANT TISSUE
CULTURE
Plant tissue culture includes two major methods:
• (A) Type of in vitro growth-callus and suspension
cultures.
• (B) Type of explant— single cell culture, shoot
and root cultures, somatic embryo culture,
meristem culture, anther culture and haploid
production, protoplast culture and somatic
hybridisation, embryo culture, ovule culture,
ovary culture, etc.

9. Environmental Conditions
• There are three important aspects in vitro
culture
1) Nutrient medium
2) Aseptic condition
3) Aeration of the tissue

10. Nutrient Medium
• Medium depends upon the type of plant tissue
or cell used for culture
• Generally nutrient consist of
 inorganic salts (both micro & macro elements)
 a carbon source (usually sucrose)
 Vitamins (eg. nicotinic acid, thiamine, pyridoxine
 Amino acids (eg. arginine)
 Growth regulators (eg. auxins)
• An optimum pH (5.7) is also vary important

11. Aseptic Condition
• Nutrient medium contains sugar which
increases growth of microbes
• These microbes compete with growing tissue
and finally kill it.
• It is important to maintain aseptic condition.
• Sterilization is very important to stop the
growth of microbes.

12. Aeration of the Tissue
• Proper aeration of the cultured tissue is also an
important aspect of culture technique.
• It is achieved by occasionally stirring the
medium by stirring or by automatic shaker.

13. Types of Plant Tissue
Culture

14. Callus Culture
• In Callus culture, cell division in explant forms a
callus.
• Callus is irregular unorganized and undifferentiated
mass of actively dividing cells.
• Darkness & solid medium gelled by agar stimulates
callus formation.
• The medium contains the auxins and BAP (Benzyl
amino purines). Both are growth regulators
( Hormones).
• This stimulates cell division in explant.
• Callus is obtained within 2-3 weeks.

15. Callus culture and Sub culture

16. Maintenance
• After sufficient time of callus growth on the
same medium following changes will occur :
 Depletion of nutrient in the medium
 Gradual loss of water
 Accumulation of metabolic toxins
• Hence for maintenance of growth in callus it is
necessary to subculture the callus.
• Subculture should be repeated after 4-5 weeks

17. Single Cell Culture
• As stated earlier, cells derived from a single
cell through mitosis constitute a clone and the
process of obtaining clones is called cloning
(asexual progeny of a single individual make
up a clone). There are two popular techniques
for single cell culture.

18. Root tip culture
• Tips of the lateral roots are sterilized, excised and
transferred to fresh medium.
• The lateral roots continue to grow and provide
several roots.
• After 7 days that are used to initiate stock or
experiment.
• Thus the root material derived from a single
radical.
• Such genetically uniform root cultures are
referred to as a clone of isolated roots.

19. Leaves culture
• Leaves (800µm) may be detached from shoots,
surface sterilized and place in healthy
condition for long period.
• Growth rate in the culture depends on their
stages of maturity at excision.
• Young leaves have more growth potential then
the nearly mature ones.

20. Shoot tip culture
• The excised shoot tips (100-1000µm long) of
many plant species can be cultured on
relatively simple nutrient media.
• This media must contains growth hormones
and will often form roots and develop into
whole plants.

21. Complete flower culture
• Flowers (2days after pollination) are excised,
sterilized by immersion in 5% calcium
hypochloride, washed with sterilized water.
• Transfer this to culture tubes containing an
agar medium.
• Fruits, which develop are smaller than their
natural counterpart, size can be increases by
supplementing the medium with appropriate
combination of growth hormones.

22. Anther Culture
• Young flower buds are removed from the plant
& surface sterilized.
• The anthers are then excised and transferred to
an appropriate nutrient medium.
• The plantlet are formed after 4-5 weeks of
inoculation.
• Many plantlets are produced from the single
anther.

23. Pollens culture
• Pollen grains are removed from the anther.
• Anthers are placed in a 5ml liquid medium in
petri dish.
• Petri dishes containing the pollen grains in the
culture media are sealed with parafilm &
incubated at 28ºC in dark for 14 days.
• 3-4 weeks may be required to obtain haploid
plantlets.

24. Application of tissue culture
• Tissue culture is used to conserve the rare
species in the forest.
• A plant breeder may use tissue culture to
screen cells rather than plants.
• Large-scale growth of plant cells in liquid
culture as a source of secondary products, like
recombinant proteins used as
biopharmaceuticals.

25. • To cross distantly related species by protoplast
fusion.
• Help in crop improvement.
• Creation of additional genetic variation.
• Selection of plants resistant to toxins, viruses
etc.
• Suitable for breeding of tree species.
• Production of many plants that are clone to
each other.
• Production of disease free plants.

26. • Tissue culture have a advantage of time sever,
With the help of root culture plants are ready to
grow which save time.

27. POLYPLOID
• Polyploidy refers to when an organism has more
than two complete sets of chromosomes.
• This condition is frequently found in plants.
Polyploids can be divided into types based on the
number of chromosomes they carry
1. Triploids (three sets)
2. Tetraploids (four sets)
3. Pentaploids (five sets)
4. Hexaploids (six sets)
5. Octoploids (eight sets)
6. Decaploids (ten sets)
7. Dodecaploids (twelve sets)

28. Importance of Polyploids
1) Identify genetic origin of crops.
2) Generate new plant genotypes and species.
3) Enable introgression of genes from related
species.
4) Self fertilization.
5) Heterosis ( crossbred plant species having superior
qualities than those of either of its parents).
6) Complete the no of chromosomes having
uneven sets of chromosomes.

29. 7) A plant may be made sterile by developing a
triploid - fertile tetraploids and diploids are
hybridized to get triploids.
8) Polyploidy can be used to obtain seedless fruits,
such as the seedless watermelon. Another example
of successful polyploidy is the banana, which is a
triploid, seedless and sterile.
9) As with hybrids, polyploidy have been observed
to be bigger and more robust than their diploid
parents. This may include bigger leaves, flowers and
fruits, as well as an increase in height. An example
is the strawberry plant, diploid varieties of
strawberry have smaller fruit with less yields,
whereas the tetraploid varieties have bigger and
better yields. The polyploids can also be more
disease and pest resistant.

30. Seedless Watermelon Strawberry (Heterosis)

31. Mutation
• Mutation is the inheritable change in the
characters of organisms.
• Mutation can occur spontaneously in nature or
they may be induced experimentally.
• Mutation can be beneficial or harmful to an
individual.

32. Importance of Mutation
• Evolution of new and better food plants having
good desirable characters such as higher yield
and greater resistance.
• Production of new species.
• Agricultural revolution.
• Mutation breeding often provide solution for
some specific plant breeding problems.
• Can produce improved verity of plants than the
traditional verity.
• Less time consuming.

33. • Improvement in genetic characters so that the
new species give desirable effects.
• To increase crop yield.

34. Hybridization
• Hybridization is the process of crossing two
genetically different individuals to result in a
third individual with a different one.