banana tissue culture

The propagation of a plant by using a plant part or single cell or group cell in a test tube under very controlled and hygienic conditions is called "Tissue Culture".

Banana is a globally important fruit crop with 97.5 million tones of production. In India it

supports livelihood of million of people. With total annual production of 16.91 million

tones from 490.70 thousand ha., with national average of 33.5 T/ha. Maharashtra ranks

first in production with 60 T/ha. Banana contributes 37% to total fruit production in India.

Banana is one of the major and economically important fruit crop of Maharashtra.

Banana occupy 20% area among the total area under crop in India. Maharashtra ranks

second in area and first in productivity in India. Jalgaon is a major Banana growing

district in Maharashtra which occupy 50,000 hectares area under Banana. But most of

Banana is grown by planting suckers. The technology development in agriculture is very

fast, it results in developing Tissue Culture Technique.

Banana is basically a tropical crop, grows well in temperature range of 13ºC – 38ºC with RH regime of 75-85%. In India this crop is being cultivated in climate ranging from humid tropical to dry mild subtropics through selection of appropriate varieties like Grandnaine. Chilling injury occurs at temperatures below 12ºC. The normal growth of the banana begins at 18ºC, reaches optimum at 27ºC, then declines and comes to a halt at 38ºC. Higher temperature causes sun scorching. High velocity wind which exceeds 80 km phrs damages the crop.

Soil for banana should have good drainage, adequate fertility and moisture. Deep, rich

loamy soil with pH between 6-7.5 are most preferred for banana cultivation. Ill drained,

poorly aerated and nutritionally deficient soils are not suitable for banana. Saline solid,

calcareous soil are not suitable for Banana cultivation. Avoided soil of low laying areas,

very sandy & heavy black cotton with ill drainage.

A soil that is not too acidic & not too alkaline, rich in organic material with high nitrogen

content, adequate phosphorus level and plenty of potash are good for banana.

In India banana is grown under diverse conditions and production systems. Selection of

varieties, therefore is based on a large number of varieties catering to various kinds of

needs and situations. However, around 20 cultivars viz. Dwarf Cavendish, Robusta,

Monthan, Poovan, Nendran, Red banana, Nyali, Safed Velchi, Basarai, Ardhapuri,

Rasthali, Karpurvalli, Karthali and Grandnaine etc..

Grandnaine is gaining popularity and may soon be the most preferred variety due to its

tolerance to biotic stresses and good quality bunches. Bunches have well spaced hands

with straight orientation of figures, bigger in size. Fruit develops attractive uniform yellow

colour with better self life & quality than other cultivars.

Prior to planting banana, grow the green manuring crop like daincha, cowpea etc. and

burry it in the soil. The land can be ploughed 2-4 times and leveled. Use ratovator or

harrow to break the clod and bring the soil to a fine tilt. During soil preparation basal

dose of FYM is added and thoroughly mixed into the soil.

A pit size of 45cm x 45cm x 45cm is normally required. The pits are to be refilled with

topsoil mixed with 10 kg of FYM (well decomposed), 250 gm of Neem cake and 20 gm of

conbofuron. Prepared pits are left to solar radiation helps in killing the harmful insects, is

effective against soil borne diseases and aids aeration. In saline alkali soil where PH is

above 8 Pit mixture is to be modified to incorporate organic matter.

Addition of organic matter helps in reducing salinity while addition of purlite improves,

porosity and aeration. Alternative to planting in pits is planting in furrows. Depnding on

soil strata one can choose appropriate method as well as spacing and depth at which

plant is required to be planted.

Sword suckers weighing approximately 500-1000 gm are commonly used as propagating

material. Suckers generally may be infected with some pathogens and nematodes.

Similarly due to the variation in age and size of sucker the crop is not uniform, harvesting

is prolonged and management becomes difficult.

Therefore, in-vitro clonal propagation i.e. Tissue culture plants are recommended for

planting. They are healthy, disease free, uniform and authentic. Properly hardened

secondary seedlings are only recommended for planting

True to the type of mother plant under well management.

Pest and disease free seedlings.

Uniform growth, increases yield.

Early maturity of crop - maximum land use is possible in low land holding country like India.

Round the year planting possible as seedlings are made available throughout the year.

Two s uccessive ratoons are possible in a short duration which minimizes cost of

cultivation.

No staggered harvesting.

95% - 98% plants bear bunches.

New varieties can be introduced and multiplied in a short duration.

Planting of tissue culture Banana can be done throughout the year except when

the temperature is too low or too high. Facility of drip irrigation system is

important. There are two important seasons in Maharashtra, India;

Mrig Baug (Kharif) Month of planting June - July.

Kande Baug (Rabi) Month of planting October - November.

Crop Geometry

Traditionally banana growers plant the crop at 1.5m x 1.5m with high density,

however plant growth and yields are poor because of competition for sunlight.

Various trials are conducted at Jain Irrigation System R&D farm with Grandnaine

as cultivar. And then suitable spacing of 1.82m x 1.52m is being recommended, it

accommodates 1452 plants per acre (3630 plants per hectare) keeping row

direction North-South with wide spacing 1.82m between the rows. The region like

north India, coastal belt and where humidity is very high and temp falls down upto

5-7ºC, the planting distance should not be less than 2.1m x 1.5m.

Polybags is separated from the plant without disturbing the root ball of the plant

and then plants are planted in the pits keeping the pseudo-stem 2cm below the

ground level. Soil around the plant is gently pressed. Deep planting should be

avoided.

Banana, a water loving plant, requires a large quantity of water for maximum

productivity. But Banana roots are poor withdrawal of water. Therefore under Indian

condition banana production should be supported by an efficient irrigation system like

drip irrigation.

Water requirement of banana has been worked out to be 2000mm per Annum.

Application of drip irrigation and mulching technology has reported improved water use

efficiency. There is saving of 56% of water and increasing yield by 23-32% under drip.

Irrigate the plants immediately after planting. Apply sufficient water and maintain field

capacity. Excess irrigation will lead to root zone congestion due to removal of air from

soil pores, thereby affecting plant establishment and growth. And hence drip method is

must for proper water management in Banana

Month (Maug Baug) Qty. (lpd.) Month (Kande baug) Qty. (lpd.)

June 06 October 04-06

July 05 November 04

August 06 December 04

September 08 January 06

October 10-12 February 08-10

November 10 March 10-12

December 10 April 16-18

January 10 May 18-20

February 12 June 12

March 16-18 July 12

April 20-22 August 14

May 25-30 September 14-16

Banana requires high amount of nutrients, which are often supplied only in part by the soil. Nutrient requirement has been worked out on all India basis is to be 20 kg FYM, 200gm N; 60-70gm P; 300gm K/plant. Banana requires heavy nutrition. Banana crop requires 7-8 Kg N, 0.7- 1.5 Kg P and 17-20 Kg K per metric tonne yield. Banana responds well to application of nutrients. Traditionally farmers use more of urea and less of phosphorous and potash.In order to avoid loss of nutrients from conventional fertilizers i.e. loss of N through leaching, volatilization, evaporation and loss of P and K by fixation in the soil, application of water soluble or liquid fertilizers through drip irrigation (fertigation) is encouraged. A 25-30% increase in yield is observed using fertigation. Moreover, it saves labour and time and the distribution of nutrients is uniform.

The fertilizer schedule for tissue culture banana variety Grand Naine both in solid and

water soluble form is given in the tables below:

Solid fertilizer schedule for Grand Naine Banana

Total nutrient requirement

N - 200 gm/plantP - 60-70

gm/plantK - 300 gm/plant

Total quantity of fertilizer required per acre (Spacing 1.8 x 1.5 m;

1452 plants)

Urea (N) SSP (P) MOP (K)

431.0 375.0 500 gm/plant

625.0 545.0 726 kg/acre

Period Application Source Quantity (gm / plant)

At the time of Planting S.S.P. 100

M.O.P. 50

10th Day after planting Urea 25


S.S.P. 100

M.O.P. 50

Micronutrient 25

MgSO4 25

Sulphur 10


S.S.P. 100

M.O.P. 50


S.S.P. 100

M.O.P. 50

Micronutrient 25

Sulphur 30

MgSO4 25


M.O.P. 100


M.O.P. 100


M.O.P. 60


M.O.P. 60


M.O.P. 60


M.O.P. 60


M.O.P. 60

Schedule is directive only and may change according to planting season and soil fertility

status (Soil analysis)

SSP = Single Super Phosphate, MOP = Muriate of Potash.

Water Soluble Solid fertilizers

Schedule of water soluble fertilizer application.

PeriodGrade

Qty. per 1000 plants

(Kg)

every 4th day basis

Total Qty.

(Kg.)

After planting upto 65 days

Urea 4.13 82.60

12:61:00 3.00 60.00

00:00:50 5.00 100.00

65 to 135 days

Urea 6.00 120.00

12:61:00 2.00 40.00

00:00:50 5.00 100.00

135 to 165 daysUrea 6.50 65.00

00:00:50 6.00 60.00

165 to 315 daysUrea 3.00 150.00

00:00:50 6.00 300.00

Schedule is directive only and may change according to planting season and soil fertility

status (soil analysis).

The Root system of banana is superficial and easily damaged by cultivation, use of

intercrop which is not desirable. However short durational crops (45-60 days) like mung,

cowpea, daincha are to be considered as green manuring crops. Crops from

cucurbitaceous family should be avoided as these carry viruses.

Weeding

Spraying of Glyphosate (Round up) before planting at the rate of 2 lit/ha is carried out to

keep the plantation weed free. One or two manual weedings are necessary.

Micronutrient Foliar Spray

Combined foliar application of ZnSo4 (0.5%), FcSo4 (0.2%), CuSo4 (0.2%) and H3Bo3

(0.1%) can be adopted to improve morphological, physiological and yield attributes of

banana. The micronutrient spray solution is prepared by dissolving the following in 100

lit. of water.

Zinc sulphate 500

gm

For every 10 litre of mixture 5-10ml of sticker

solution such as

Teepol should be added before spraying.

Ferrom

sulphate

200

gm

Copper

sulphate

200

gm

Boric acid100

gm

There are operation specific to banana crop which influence productivity and quality.

Desuckering

Removal of unwanted suckers is a critical operation in banana for reducing internal

competition with mother plant.

Desuckering should be done regularly until shooting. However in areas where ratoon is

also taken for the second crop, a follower is allowed after inflorescence has appeared and

this should be managed that planting space is not disturbed. Follower should be opposite

to the inflorescence. It should not be far apart from the main plant.

Deflowering

It consists of removal of the withered style and perianth. This is generally not practiced.

Therefore, they remain attached to the fruit bunch & then removed after harvesting

which is damaging to the fruits. It is therefore suggested that you remove them just after

flowering.

Pruningof leaves

Rubbing leaves damages the fruit, therefore, such leaves should also be pruned during

regular check. Older leaves and infected leaves also be pruned as required. Green leaves

should not be removed.

Earthing up

Keep the soil loose by harrowing from time to time. Earthing up should be done at 3-4

months after planting i.e. raising the soil level around the base of the plant by 10-12”. It

is better to prepare a raised bed and keep the drip line on bed 2-3” away from the plant.

It also helps to protect plants from wind damage and production losses to some extent..

Removal of male buds

(Denavelling) Removal of male buds helps fruit development and increases bunch

weight. Male buds are removed from the last 1-2 small hands with a clean cut keeping a

single finger in the last hand.

Bunch Spray

Spray of monocrotophos (0.2%) after emergence of all hands takes care of the thrips.

Thrips attack discolors the fruit skin and makes it unattractive.

Bunch Covering

Covering bunch using dried leaves of the plant is economical and prevents the bunch

from direct exposure to sunlight. Bunch cover enhances quality of fruit. But in rainy

season this practice should be avoided.

Sleeving of bunch is done to protect fruits against dust, spray residue, insect and birds.

For this blue plastic sleeves are preferred. This also increases temperature around

developing bunch and helps in early maturity.

Dehandling of false hands of bunch

In a bunch there are some incomplete hands which are not fit for quality produce. These

hands should be removed soon after bloom. This helps in improving the weight of other

hands. Sometimes the hand just above the false hand is also removed.

Propping

Due to heavy weight of bunch the plant goes out of balance and the bearing plant may

lodge and production and quality are adversely affected. Therefore they should be

propped with the help of two bamboos forming a triangle by placing them against the

stems on the leaning side. This also helps in uniform development of bunch.

A large number of fungal, viral and bacterial diseases and insect pests and nematodes

infest the banana crop and reduce production, productivity and quality. Summary details

of major pest and diseases of banana along with control measures are given herewith:

Sno

.Name Symptoms Control measures

Pest

i) Rhizome

weevil(Cosmopolites

Sordidus)

a) Large creates

network of galleries in

rhizome and weakens

the plant.

a) Use healthy planting

material

b) Sanitation in orchard

c) Trapping of adult

weevils using pseudostem

or rhizome pieces and

d) Soil application of

carbufuran @.2gm/plant

ii) Pseudostem

weevil(Odaiporous

longicolis)

a) Small holes on

pseudostem with

exudation of

transparent gummy

substance

a) Management approach

is identical to rhizome

weevil

b) Existence tunneling

in leaf sheath and

inner core of the stem

b) Secondly, injection of

lime solution

(Monocrotophos 150 ml in

350 ml water) using stem

injector 4 ft. above the

ground level at 30º angle

is recommended.

c) Abortion of bunches c) Use longitudinal split

(30cm length) or disc on

stump traps @ 100/ha.

Keep the split portion of

tray facing the ground.

Collected weevils are then

killed.

iii) Thrip

s(Chaetanaphotrips &

signipennis &

Heliaothrips

kodaliphilus)

a) They scrap from

attacked plant organs

and render them

brown and discolored

especially the fruits.

a) Spray or inject

Monocrotophos @ 0.05%

on the inflorescence

before the unfurling of top

most bract.

iv) Fruit scarring

battle(Besilepta

subcostatum)

a) Adults feed on

tender unfolded leaves

and fruits and cause

scarring of skin

a) Sanitation spray of

0.05% moncrotophos or

0.1% carbaryl on the

heart of the plants

immediately after the

emergence of new foliage

and during fruiting season

is recommended.

b) Plant losses its

vigour and quality of

bunch is poor)

v) Aphids (Pentalonia

nigronervosa)

a) They are vecturs of

banana bunchy top

visus (BBTV) and can

be seen as

congregation under

the leaf base of

pseudo stem

a) Spray of 0.1%

monocrotophos or 0.03%

phosphonidon on the

leaves is effective

vi) Nematodes a) Stunted growth a) Apply corbofuron @40

gm per plant at planting &

4 month after planting.

b) Small leaves

c) Cutted roots b) Use neem cake as

organic manure.

d) Purple black lesions

on roots and their

splitting.

c) Use merigold as trap

crop.

Fungal Diseases

vii) Panama wilt(Furarium a) Yellowing of old a) Cultivation of resistant

oxysporium) leaves progressing cultivar towards younger

leaves. (Covendish group)

b) Affected leaves

collapse near petiole

and hang.

b) Trim and treat the

suckers in 0.1% Bavistine

before plant.

c) Pseudo stem

splitting is common.

c) Apply bioagents like

trichoderma and

Pseudomonas

fluorescence with organic

manure

d) Reddish brown

discoloration in cross-

section of root &

rhizome

d) Keep good drainage

and apply lot of organic

manure in field.

viii Head rot (Erwinia

carotovora)

a) Rotting of collar

region and epinasty of

leaves)

a) Use healthy planting

material

b) On pulling out of

affected plant, the

plant topples from the

collar region leaving

the corn with root in

soil

c) On opening up of

collar region of

affected plants,

yellowish to reddish

ooze can be seen.

b) Drench plants with

0.1% Emison followed by

another drenching after 3

months.

d) In early stage of

infection, dark brown

or yellow, water

soaked areas in critical

region which may

decay to form cavities

surrounded by dark

spongy tissues.

c) Avoid planting in rocks

and in poorly drained

soils.

ix) Sigatoka leaf spot a) It is characterized a) Remove infected leaves

(Mycospharella spp) by small lesions on the

leaves, the lesion

become pale yellow to

greenish yellow

streaks visible from

both the surfaces of

leaf

and destroy

b) Thereafter linear

brownish to blackish

streaks appear.

c) The centre of the

streak eventually dries

up and give

appearance of eye

spot.

b) Keep proper drainage

and avoid water logging.

d) Some times

premature ripening is

observed

c) Spray dithane M-45

(1250 g/ha) or Bavistine

500 g/ha.

Viral Diseases

i) Banana Bunchy Top

Virus(BBTV)

a) Appearance of

irregular, dark green

'Morse code' streaks

along secondary veins

on leaves on underside

of the leaves.

a) Use virus free planting

material i.e. Tissue

Culture.

b) Survey and eradicate

infected plants regularly.

b) Leaf size is reduced

and leaves remains

abnormally erect,

brittle and results.

c) Control insect vectors

especially aphids and

mealy bugs.

d) Indexing should be

followed in the case of

mass multiplication

c) Leaves short, close

to each other, and

bunched at the top

e) Prohibit movement of

any plant part from

diseased area to healthy

area.

d) The tips of the

bracts in male buds

f) Use resistant cultivar.

have greenish.

e) Virus is spread

through aphids.

g) Avoid growing of

alternate lost as mixed

crop or in near by areas.

ii) Banana Mosaic

Virus(BMV)

a) Chlorosis with mild

chlorotic streaks along

the veins they never

turn necrotic as in BSV.

a) Elimination of affected

plants and maintenance of

disease free plantation

through the use of disease

free planting material i.e.

Tissue culture seeding.

iii) Banana Bract Mosaic

Virus(BBMV)

a) Presence of spindle

shaped pinkish to

reddish streaks on

pseudo stem, mid ribs,

petioles and lamina.

a) Use of disease free

planting material i.e.

Tissue culture seeding.

iv) Banana Streak Virus(BSV) a) Presence of

inconspicuous chlorotic

flecking to small lethal

systematic necrosis,

and includes yellow,

brown and black

streaking, cigar leaf

necrosis, based

pseudo stem splitting

internal internal

pseudo stem necrosis

and formation of small

deformed bunches.

a) Use of disease free

planting material i.e.

Tissue culture seedings.

Banana should be harvested at the physiological maturity stage for better post harvest

quality. The fruit is climacteric and can reach consumption stage after ripening operation

Maturity indices

These are established on the basis of fruit shape, angularity, grade or diameter of the

median figure of the second hand, starch content and number of days that have elapsed

after flowering. Market preferences can also affect the decision for harvesting a slight or

full mature fruit.

Removal of bunch

Bunch should be harvested when figures of second hand from top are 3/4 rounded with

the help of sharp sickle 30cm above the first hand. Harvest may be delayed upto 100-

110 days after opening of the first hand. Harvested bunch should generally be collected

in well padded tray or basket and brought to the collection site. Bunches should be kept

out of light after harvest, since this hastens ripening and softening.

For local consumption, hands are often left on stalks and sold to retailers.

For export, hands are cut into units of 4-16 fingers, graded for both length and girth, and

carefully placed in polylined boxes to hold different weight depending on export

requirements.

Post harvest operations

At collection site injured and over mature fruits are discarded and for local market bunches should be delivered through lorries or wagons. However, for more sophisticated and export market where the quality is predominant, bunches should be dehanded, fruits are cleared in running water or dilute sodium hypochlorite solution to remove the latex and treated with thiobendasole; air dried and graded on the basis of size of fingers as already stated, packed in ventilated CFB boxes of 14.5 kg capacity or as per requirement with polythene lining and pre-cooled at 13-15ºC temperature and at 80-90% RH.Such material should than be sent under cool chain at 13ºC for marketing

YIELD

The planted crop gets ready for harvest within 11-12 months of planting. First ratoon

crop would be ready by 8-10 month from the harvesting of the main crop and second

ratoon by 8-9 months after the second crop.

1. Banana CultureR.SathesStudentDepartment of Plant Biology and Bio technologyLoyola College

2. What is Tissue Culture?Tissue culture is collection experimental methods of isolation and inoculation of organs, tissues

and cell in an artificial medium under in-vitro aseptic condition.Micro propagation is vegetative propagation of plant using plant tissue culture. This also known as direct differentiation.

3. Banana Cultivation in IndiaBanana is a globally important fruit crop with 97.5 million tones of production. In India it supports livelihood of million of people. With total annual production of 16.91 million tones from 490.70 thousand ha., with national average of 33.5 T/ha.Maharashtra ranks first in production with 60 T/ha. Banana contributes 37% to total fruit production in India.Banana occupy 20% area among the total area under crop in India. Jalgaon is a major Banana growing district in Maharashtra which occupy 50,000 hectares area under Banana. But most of Banana is grown by planting suckers. The technology development in agriculture isvery fast, it results in developing Tissue Culture Technique.

4. Requirement for Banana Growth

5. Agro ClimateBanana is basically a tropical crop, grows well in temperature range of 13ºC – 38ºC with RH regime of 75-85%. In India this crop is being cultivated in climate ranging from humid tropical to dry mild subtropics through selection of appropriate varieties like Grandnaine. The normal growth of the banana begins at 18ºC, reaches optimum at 27ºC, then declines and comes to a halt at 38ºC. Higher temperature causes sun scorching. High velocity wind which exceeds 80 km phrs damages the crop.

6. SoilSoil for banana should have good drainage, adequate fertility and moisture. Deep, rich loamy soil with pH between 6-7.5 are most preferred for banana cultivation. Ill drained, poorly aerated and nutritionally deficient soils are not suitable for banana.Saline solid, calcareous soil are not suitable for Banana cultivation. Avoided soil of low laying areas, very sandy & heavy black cotton with ill drainage.A soil that is not too acidic & not too alkaline, rich in organic material with high nitrogen content, adequate phosphorus level and plenty of potash are good for banana.

7. VarietiesIn India banana is grown under diverse conditions and production systems. Selection of varieties, therefore is based on a large number of varieties catering to various kinds of needs and situations.Dwarf Cavendish.Robusta. MonthanPoovanNendranRed bananaNyaliSafedVelchiBasaraiArdhapuriRasthaliKarpurvalliKarthali Grandnaine

Grandnaine is gaining popularity and may soon be the most preferred variety due to its tolerance to biotic stresses and good quality bunches.

8. Grandnaine variety

9. Planting MaterialSword suckers weighing approximately 500-1000 gm are commonly used as propagating material.

10.

11. Why tissue culture?Suckers generally may be infected with some pathogens and nematodes. Similarly due to the variation in age and size of sucker the crop is not uniform, harvesting is prolonged and management becomes difficult.Therefore, in-vitro colonel propagation i.e. Tissue culture plants are recommended for planting. They are healthy, disease free, uniform and authentic. Properly hardened secondary seedlings are only recommended for planting

12. Advantages of Tissue Culture Planting MaterialTrue to the type of mother plant under well management.Pest and disease free seedlings.Uniform growth, increases yield.Early maturity of crop - maximum land use is possible in low land holding country like India.Round the year planting possible as seedlings are made available throughout the year.Two successive ratoons are possible in a short duration which minimizes cost of cultivation.No staggered harvesting.95% - 98% plants bear bunches.New varieties can be introduced and multiplied in a short duration.

13.

14. MicropropagationOf Banana

15. Initiation of shoot culturesShoot cultures of banana start conventionally from any plant part that contains a shoot meristem, i.e. the parental pseudo stem, small suckers, peepers and lateral.For rapid in vitro multiplication of banana, shoot tips from young suckers of 40-100 cm height are most commonly used as explants. From the selected sucker a cube of tissue of about 1-2 cm³ containing the apical meristem is excised.

16. The optimal size of the explants depends on the purpose. For rapid multiplication, a relatively larger explants (3-10 mm) is desirable despite its higher susceptibility to blackening and contamination. When virus or bacteria elimination is needed, meristem-tip culture is the preferred option. The explants is then further reduced in size (0.5-1 mm length), leaving a meristematic dome with one or two leaf initials. Meristem cultures have the disadvantage that they may have a higher mortality rate and an initial slower growth.

17. The suckers for culture is prepared and taken into the lab.They are soaked in Bavistin for 18 hours in order to remove the fungus and fungal spores.In lab first they are washed in running water.Then they are Dipped into detergent water (teepol) containing for one hour.After that they are taken and washed in tap water.Washed suckers are taken to LAF chamber and further sterilization is done.

18. In LAF chamber the suckers are sterilized using mercury chlorideTwo different concentrations are used for sterilization purpose.First the sucker sterilized using 0.12% of HgCl2. The sucker put into the bottle containing HgCl2 Shake well for 2 min.After that HgCl2is removed and the sucker washed using distilled water. At first the distilled water added and the bottle shaked for 1 min. then the water removed and fresh distilled water added shaked for another one min. The water removed.This step repeated under following timings 2min,3min,5min and 12 min.

19. After 1st sterilization a layer of the sucker removed carefully.The suckers are again sterilized using 0.1% HgCl2 for 5 min.After that they are washed thoroughly with distilled water in following timings 1min,1min,2min,3min,5min,12min. Finishing the above process another layer of sucker is removed.The suckers are ready for inoculation.

20. SuckersLayer removal1st Layer2nd LayerInner Layer2nd sterilization1st sterilizationRemoval of a layerRemoval of a layer

21. Sterilization process

22. Medium used for Banana cultivationFor banana micro propagation, MS-based media are widely adopted. Generally, they are supplemented with sucrose as a carbon source at a concentration of 30-40 g/l. Media are poured in a glass bottle where suckers are propagated.Usually two types of growth regulators used, a cytokinin and an auxin, are added to the banana growth medium. Their concentration and ratio determines the growth and morphogenesis of the banana tissueIn most banana micro propagation systems, semi-solid media are used. As a gelling agent agar (5-8 g/l) is frequently added to the culture medium.

23. Problem of banana propagationBanana tissue cultures often suffer from excessive blackening caused by oxidation of polyphenolic compounds released from wounded tissues. These undesirable exudates form a barrier round the tissue, preventing nutrient uptake and hindering growth. Therefore, during the first 4-6 weeks, fresh shoot-tips are transferred to new medium every 1-2 weeks. Alternatively, freshly initiated cultures can be kept in complete darkness for one week.Antioxidants, such as ascorbic acid or citric acid in concentrations ranging from 10-150 mg/l, are added to the growth medium to reduce blackening, or the explants are dipped in antioxidant solution (cysteine 50 mg/l) prior to their transfer to culture medium.

24. Transfer to Growth room Banana shoot-tip cultures are incubated atAn optimal growth temperature of 28 ± 2°C In a light cycle of 12-16 h with a photosynthetic photon flux (PPF) of about 60 µE/m2s1.Air condition will be working all the time to provide needed temperature. Which also provide clean dust free environment.

25. Growth Room

26. Inoculate sucker & Sucker after 2 weeks

27. Arise of shoots from suckers

28. Sub Culturing of BananaAfter 2 weeks the suckers will become greenish in colour.2 weeks after that multiple shoot will arise from the base of the suckers.The shoots are cut at the base separated and placed in a fresh medium.In each bottle three shoots are inoculated.After a week multiple shoots arise from the inoculated shoot. Again they are separated and placed in a new fresh medium.

29. Sub Cultured Banana

30. Sub culturing and inoculated cultures

31. The sub culturing is done until the require amount of plant needed.The shoots are every day checked for contamination and the contaminated shoots are transferred to a fresh medium.Meanwhile a set of well grown healthy shoots are taken for rooting.

32. Well grown healthy shoots

33. Rooting of Banana ShootsRooting of banana shoot is done in bottle containing charcoal medium.For rooting IAA Used as growth regulator (for commercial purpose).Medium without hormone gives good results.It will take 2 weeks for rooting and fresh roots are arise at the base of the shoot.

34. Rooted Plantlet

35. Different Stages Of Banana Culture

36. Acclimatization of banana plantlet Rooted plantlet are kept in basal medium for certain time.Then there are taken for acclimatization process where they gradually trained to the in-vivo temperature and light.There are two stages in hardening processPrimary HardeningSecondary HardeningThe hardened plantlets are carefully maintained in green house.

37. Plates used for primary Hardening

38. Secondary Hardening

39. Transporting the plantlets.The plantlets after acclamatization should be transported to the required place. Normal transportation is done where the plants are placed and grown in Plastic Bags.Hence the well grown plants removed to provide the space in green house for the next cycle of plants and also to lower the cost of storage.

40. Scope of banana CultureBanana have lot of useful properties which built the pathway for banana growers.Normal banana suckers which obtained directly from the plantation grows in 12 to 15 month where PTC plantlet will grow in 10 months.The suckers are collected for PTC are healthy and high yielding so the daughter plantlets have the same character of the mother plant.

The banana is a short term crop so it provides immense opportunity to produce PTC plants through out the year.Care full measures should be taken in growing PTC banana which can give a good job opportunity and income for the producers.

Thus over a period to 28-30 months, it is possible to harvest three crops i.e. one main

crop and two ratoon crops. Under drip irrigation combined with Fertigation yield of

Banana as high as 100 T/ha can be obtained with the help of tissue culture technique,

even similar yield in the ratoon crops can be achieved if the crop is managed well.

banana tissue culture

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