steps & procedure of micro propagation

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MICRO PROPAGATION IN BANANA
AND
POMEGRANATE

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Created by…………….
NAME:- digambar hole
REG NO:- 2009/063
Department Of Botany & Plant Tissue
Culture(ELBOT)
College Of Agriculture Pune-05

4. Introduction.
Why do micropropagation?
Advantages of micropropagation.
General steps of micropropagation.
Varieties grown in India.
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content

5. Banana is a globally important fruit crop with 97.5 million tones
of production.
India produces about 26.217 million tones of Banana from an
area of 0.709 Mha., with an average productivity of 37.0 mt/ha.
Micropropagation is refers to rapid asexual in vitro
multiplication of a desired plant of banana.
In India only 7 percent area under the tissue culture plantlets
and remaining 93percent propagated by sucker.
New entrepreneurs having sound scope in production of banana
micropropagated plantlets.
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INTRODUCTION

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Leading Banana Producing Countries
Source: National Horticulture Board (NHB) 2011-12

7. IndianProductionof BananaState wise Production of Banana in India
Source: National Horticulture Board (NHB) 2011-12
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What is MICROPROPAGATION ?
A whole plant can be regenerated from a small tissue or
plant cells in a suitable culture medium under controlled
environment. The plantlets so produced are called tissue-
culture raised plants.

9. 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.
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Why do micropropagation ?

10. 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).
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Cont.…

11. 1.Rapid multiplication
2.Requirement of limited mother stock
3.Product uniformity
4.Season independent production
5.Agronomic advantages
6.Plant exchange
7.High returns
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ADVANTAGES OF micropropagation

12. 12Source : Cadila pharmaceuticals limited

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Stage 0 : Selection and maintenance of stock plants for culture
initiation
EXPLANT
SHOOT TIPS FROM YOUNG
SUCKER
APICAL MERISTEM (1-2 Cm3)
SURFACED STERILIZATION

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Stage- I : Initiation and establishment of aseptic culture
Source: Jain Irrigation Systems Ltd

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Initiation

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Stage –II : Multiplication of shoot

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Stage -III: Rooting

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Stage –IV: Acclimatization / Hardening

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Secondary hardening
Plants transferred to nursery bags
Kept for 6 to 8 weeks under 50% shade
Regular foliar sprays
Variation if observed is discarded
Plant ready for sale (1 feet height)/7 leaf
stage

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BANANA PLANTS READY FOR SALE

21. An ideal tissue cultureraisedplant should:
 Be 30 cm in height and have a pseudostem
circumference of 5.0-6.0 cm after 60 days of total
hardening.
 Have 4-5 photosynthetically active leaves and inter-
foliar space must be not less than 5.0 cm.
 Have approximately 25-30 more than 15 cm active
roots at the end of secondary hardening.
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Ideal tissue culture raised plant

22.  Be free from any visual symptoms of leaf spot,
pseudostem 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.
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Cont….

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Hand
Finger
Source: Jain Irrigation Systems Ltd

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Suckers Tissue
culture
Source: Jain Irrigation Systems Ltd

25. Sl.
No
Parameters TC
propagated
bananas
Sucker
propagated
bananas
Comparison
percentage
1 Mean yield (bunches/ha) 2,663 2,416 -
2 Mean price received (/bunch) 94.47 76.42 -
3 Value of main product (/ha) 2,51,573 184,630 -
4 Value of by-product (/ha) 1729 2,518 -
5 Gross income (/ha) 2,53,302 187,149 26%
6 Total expenses (/ha) 1,41,040 108,294 23%
7 Net income (/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 -
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Comparative income from tissue-culture and sucker-propagated
bananas
Source: Hanumantharya et al. (2009)

26. State Varieties grown
Maharashtra Grand naine, safed velchi, (AB), Dwarf Cavedish, Shindhuri,
Ardhapuri, Lalvelchi (Mysore), Rajeli (Plantain).
Gujarat Grand naine, Dwarf Cavedish, Lactan, Harichal, Gandevi Selection.
Tamil nadu Grand naine, matti, sevvazhai (red banana), ney poovan, robusta
poovan (mysore), sthali (silk), virupakshi (pome), pachanadan,
nedran, sakkai.
Andhra Pradesh Grand naine, Dwarf Cavedish, robusta.
Assam Jahaji, borjahaji, manjahaji, robusta, honda, chenichampa, alpan,
malbhoj (silk), jatikol (mysore) manohar, bhimkol.
Bihar Dwarf Cavedish, chinai, chenichampa, alpan (mysore), malbhoj
(silk), gauria .
Karnataka Grand naine, Dwarf Cavedish, poovan (mysore), robusta , elakki
bale.
Kerala Njali poovan, red banana, nendran (plantain), palayankondan
(mysore) poovan silk, monthan.
West bengal Amrit sagar, giant governor, lacton, mortman (silk), champa
(mysore)
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Varieties grown in India

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MICROPRAPOGATION
in
POMEGRNATE

28. SELECTION OF EXPLANT FOR
MICROPRAPOGATION OF POOMEGRANATE
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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 resedues of bavistin

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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.
After follow all techniques of sterilization it used in
micro propagation.

30. Culturemedia for pomegranate
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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.

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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.

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

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By this method
Micro propagation
is
carried out in
Banana
&
pomegranate

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