Micropropagation is a technique used to rapidly multiply plant materials under sterile conditions. The document discusses micropropagation of banana and pomegranate. For banana, tissue culture is used to produce disease-free planting materials for year-round availability and improved yields. Explants from banana suckers are sterilized and cultured on media to induce shoot formation. Shoots are then rooted and hardened for planting. For pomegranate, shoot tips are used as explants and cultured on MS media supplemented with growth regulators and compounds. This allows for mass production of true-to-type pomegranate plants.

2. 2
MICRO PROPAGATION IN DIFFERENT
CROPS
Presented by ,
ABHISHEK(BA1TAG001)
ABHISHEK.D.(BA1TAG002)
ABU DAWOOD(BA1TAG004)
ADARSHA.H.C.(BA1TAG005)
ADITHYA KUNAL(BA1TAG006)

3. INTRODUCTION :
Terminologies:
 Micro-propagation:A whole plant can be regenerated from a small
tissue or plant cells in a suitable culture medium under controlled
environment is called Micro-propagation.
 The plantlets so produced are called tissue culture raised plants.
 The size of meristem tissue used for micro-propagation is about
0.1-0.5 mm size having only one or two leaf primordia.
 Micro propagation is vegetative propagation of plant using plant
tissue culture . This also known as direct differentiation.

4.  Tissue culture: is defined as “In-vitro propagation of plants where
cells, tissues or organs of the plant body are isolated and grown
on artificial nutrient medium under aseptic conditions.”
 Totipotency : Totipotency is the ability of a single cell to divide
and produce all the differentiated cells in an organism.
 Explant :Any part of a plant taken out and grown in test tube
under sterile conditions in special nutrient media is called explant.
Ex: Root tips ,Shoot tips, Suckers ,Embryos, etc.,
 Soma clones : The plants produced by micro propagation are
genetically identical to the original plant are called Soma clones.
 Somatic Hybrids : The fusion of isolated protoplast from the cells
of two different varieties of the plant is called Somatic Hybrids.
This process is called Somatic Hybridization.
Ex: The fusion of protoplast of Tomato with Potato to form a
Hybrid called Pomato.

5. •Callus : These are the Group of undifferentiated and
unorganised mass of cells developed from the explants in tissue
culture .
•Organogenesis :It is the process of development of primordial
organs in the callus developed from explant.
Caulogenesis: Induction of shoot development from callus.
Rhizogenesis : Induction of root development from callus.
Meristemoid : Localized group of meristematic cells which give
rise to shoots or roots is called Meristemoid.
•Plasticity : It is the ability of plants to alter their metabolism,
growth and development to best suit their environment.
•Virus free plants : are raised by culturing meristems(Apical and
Axillary) ,these are transferred to land to get disease free plants.

7. •1902 - Haberlandt proposed concept of invitro cell culture
•1922 - Kolte & Robbins successfully cultured root & stem
tips respectively.
•1926 - Went discovered first plant growth hormone- Indole
acetic acid.
•1955 - Skoog & Miller discovered Kinetin as cell division
hormone.
•1957 - Skoog & Miller gave concept of hormonal control of
organ formation.
•1960-Kanta & Maheswari developed test tube fertilization
technique.

8. •1974 - Reinhard introduced biotransformation in plant
tissue cultures.
•1977 - Chilton etal. successfully integrated Ti plasmid DNA
from Agrobacterium tumefaciens in plants.
•1978 - Melchers etal. carried out somatic hybridization of
tomato & potato resulting in Pomato.
•1981 - Larkin & Scowcroft introduced the term somaclonal
variation.
•2005 - Rice genome sequenced under International Rice
Genome Sequencing Project.
•Gottlieb Haberlandt, pioneer of plant tissue culture.

9. Objectives of Micro Propagation :
1)Providing us a basic understanding of physical and
chemical requirements of cell, tissue, organ culture, their
growth and development.
2)To learn and understand a procedure that is often used to
propagate many plants of the same genetic background.
Steps of micropropagation :
Stage 0: Selection of mother plant and explant isolation.
Stage 1: Explant Establishment .
Stage 2: Shoot Multiplication .
Stage 3: Rooting of Shoots .
Stage4: Hardening and Transfer to Soil/Field.

10.  General steps :

11. BASIC REQUIREMENTS FOR MICRO-PROPAGATION :
1.A well equipped laboratory.
2.Asepsis .
3.A suitable culture medium .
4.Controlled environment .
FACTORS AFFECTING PLANT MICROPROPAGATION :
1. Growth Media.
2. Environmental Factors.
3. Explants Source.
4. Genetics.

12. IMPORTANCE OF MICRO PROPAGATION IN
AGRICULTURE :
 A single explant can be multiplied into several thousand plants in
less than a year – this allows fast commercial propagation of new
cultivars
 Once established, a plant tissue culture line can give a
continuous supply of young plants throughout the year.
 In plants prone to virus diseases, virus free explants (new
meristem tissue is usually virus free) can be cultivated to provide
virus free plants.
 Plant ‘tissue banks’ can be frozen, then regenerated through tissue
culture.
 Plant cultures in approved media are easier to export than are soil-
grown plants, as they are pathogen free and take up little space
(most current plant export is now done in this manner).

13. ADVANTAGES OF MICROPROPAGATION :
 Year-round availability of plants .
 Fast multiplication of true-to-type planting material.
 Disease-free plant production .
 Export and import of germplasm become easy requiring
minimum quarantine checks .
 Easy transport of propagation material .
 Conservation of plant diversity .
 Small space is required to maintain and multiply large number
of plants.
 Small tissue is required as an explant, hence saves the scion
wood to a great extent.
 Micro propagated plants exhibit vigorous growth and higher
yields .
 It helps in reducing the breeding cycle, through embryo rescue
and somaclonal variation .
 Production of homozygous plants.

14. DISADVANTAGES OF MICROPROPAGATION :
 Expensive laboratory equipment and service.
 No possibility of using mechanization.
 Plants are not autotrophic.
 Poor Acclimatization to the field is a common problem
(hyperhydricity).
 Mass propagation cannot be done with all crops to date.
In cereals much less success is achieved.
 Regeneration is often not possible, especially with adult
woody plant material.

15. PROBLEMS ENCOUNTERED DURING
MICRO-PROPAGATION :
 Contamination.
 Release of Phenolic Compounds.
 Variations in Tissue Culture-Raised Plants.
 Mortality in Greenhouse .
 Facilities are costly .
 Highly technical skills required .

16. MICRO PROPAGATION IN BANANA

17. INTRODUCTION :
Banana belongs to Musaceae family.
Scientific name : Musa spp.
Origin : South-East Asia.
 Second largest food-fruit crops of the world produced in
the tropical and sub-tropical regions.
Several constraints including highly season dependent and
sometimes poor quality of planting material of banana
become a limiting factor for banana propogation.
Therefore, tissue culture technology was considered an
appropriate option to overcome this problem.
The suckers are the major source of planting
material for banana and normally remain true-to-type.

18. Application of tissue culture technology to propagate banana
plants gives some advantages resulting in
disease free planting materials,
more vigorous growth,
high yields,
better quality of fruits,
earlier fruiting and
more uniform crop since they are made from selected high
yielding mother plants.

19. PLANTING MATERIAL :
•Sword suckers weighing approximately 500-1000 gm are
commonly used as propagating material.

21. PROCEDURE :
Stage 0: Selection and maintenance of stock plants for culture
initiation :
EXPLANT
SHOOTTIPSFROMYOUNG
SUCKER
APICALMERISTEM(1-2 Cm3)
SURFACESTERILIZATION

28. BANANA SUCKER TRIMMING :

29. STAGE- I : INITIATION AND ESTABLISHMENT OF ASEPTIC CULTURE :

43. STAGE –IV: HARDENING

44. Secondary hardening :
Plants transferred to nurserybags
Kept for 6 to 8 weeks under50%shade
Regular foliar sprays
Variation if observed isdiscarded
Plant ready for sale(1feet height)/7leaf
stage

45. BANANA PLANTS READY FOR SALE

46. AN IDEAL TISSUE CULTURE RAISED PLANT SHOULD:
• Be 30 cm in height and have a pseudo stem circumference of 5.0-
6.0 cm after 60 days of total hardening.
• Have 4-5 photo synthetically active leaves and inter-foliar space
must be not less than 5.0 cm.
• Have approximately 25-30 more than 15cm active roots at the
end of secondary hardening.
• Be free from any visual symptoms of Leaf spot, pseudo stem rot
and physical deformations;
• Be free from root pathogens like Erwinia, nematode lesions and
root knots.
• Random checking of roots is essential to ensure health of
plantation.
IDEAL TISSUE CULTURE RAISED PLANT :

48. Sl.
No
Parameters TC
propagated
bananas
Sucker
propagated
bananas
Comparison
percentage
1 Mean yield (bunches/ha) 2,663 2,416 -
2 Mean price received (Rs/bunch) 94.47 76.42 -
3 Value of main product (Rs/ha) 2,51,573 184,630 -
4 Value of by-product (Rs/ha) 1729 2,518 -
5 Gross income (Rs/ha) 2,53,302 187,149 26%
6 Total expenses (Rs/ha) 1,41,040 108,294 23%
7 Net income (Rs/ha) 1,12,262 78,855 30%
8 Cost of production per bunch 52.31 43.78 -
9 Net income per bunch 42.16 32.64 -
COMPARATIVE INCOME FROM TISSUE-CULTURE AND SUCKER-
PROPAGATED BANANAS :

49. MICROPRAPOGATION IN
POMEGRNATE

50. INTRODUCTION :
•Botanical name: Punica granatum L.
•Family: Punicaceae .
•Origin: Iran .
•India is one of the major pomegranates producing country.
•Salinity and drought hardy fruit crop .
•A fully matured fruit contains many of the important
nutrients, minerals, protein, fat, fiber, carbohydrate, etc.
•The fruit are rich in Fe, Ca, and antioxidant component like
phenol, pigments and tannins .

51. •The conventional method - hardwood cutting and layering
•However, it has several limitations :
- Low success
-Less availability of planting material at a time
•This results is non- availability of planting material through-
out the year .
•Micro propagation would help in overcoming difficulties of
vegetative propagation, producing true to-type plants, rapid
and mass production of planting materials .
•Tissue cultures of edible pomegranate via shoot
organogenesis, somatic embryogenesis and enhanced
auxiliary bud proliferation have been reported .

52. PROCEDURE FOR MICRO PROPAGATION IN
POMEGRANATE :
Shoot tips of about 3-5 cm long were collected from mature
trees.
Cleaned under running tap water.
And give treatment of bavistin for 30minuts for kill all
fungal infections on explant.
Wash it again 2-3 times to remove all residues of bavistin.
After washing it again wash with twin20 lab Detergent.
And take material under laminar airflow cabinet.
In laminar airflow cabinet give treatment of 0.1% mercury
chloride.

53. CULTURAL MEDIA FOR PROPAGATION :
MS-Media used for micro propagation of pomegranate we
add some other chemicals in this viz.
media supplemented with organic acids and vitamins.
Sucrose was added at 30.0 g/l and Myo-instol at 0.1g/l.
pH of the prepared media was then adjusted to 5.6- 5.8.
Agar-agar powder used for solidification of media For
establishment stage,
BAP 2mg/l & NAA is used in media.
In media also use of PVP, INISOTOL, Silver nitrate.
PVP-as phenol absorbent.

54.  After sterilization that explant transferred in media.
 Transfer explant every day in new media for remove
phenol from that explant.
 It give rooting after 12-15 days.
 After rooting subculture it for further mass
production.
 After rooting and when its potential of cultivation ends
it transfer to cocopit for primary hardening.

55. And after primary hardening it transfer for secondary
hardening in soil.
Both hardening carry out in poly houses.
After hardening it ready to sale.