Chapter No.

Introduction to Banana Tissue Culture

INTRODUCTION TO BANANA TISSUE CULTURE

Contents

3.1 Origin of Banana Fruit

3.2 Commercial Banana Cultivation
3.3 Difference Between Conventional and Scientific
Banana Farming
3.4 Brief History of Tissue Culture of Banana
3.5 Banana Tissue Culture Technology
3.6 Merits of Tissue Culture of Bananas
3.7 Scientific Plantation Method
3.8 Cost-Benefit Analysis
3.9 Marltet Potential
3.10 Banana Export
3.11 Conclusion
 INTRODUCTION TO BANANA TISSUE CULTURE

3.1 Origin of Banana Fruit

Banana is believed to have originated in South Eastern Asia with India as one
of the centers of origin apart from Indonesia, Phihppines, and Malaysia etc.
Edible banana varieties are mostly hybrids of two wild seeded species namely
Musa Acuminata and Musa Balbisiana. i

Banana is one of the popular fruits in India because of its low cost and free
availability. Banana provides a balanced diet than other fruits. Banana is
composed of mainly water and carbohydrates which provides energy (104 Cal.
Per 100 g.) It is rich in Minerals, Phosphorus and Calcium. 2

Banana ranks first in production and second in area among the fruits grown in
India, accounting for the production of 104 lakh tons annually from an area of
4 lakh hectors. Its share in total fruit production is 32 percent from 12 percent
area under fruits. The important banana growing states are Maharashtra, Gujrat,
Tamil Nadu, Karnataka, Andhra Pradesh, Kerala, Orissa, Bihar, West Bengal
and Assam.

1. Seminar on technological advancement in Banana production handling and processing

management - Souvenir 27-28 March 1999 S. Uma, B. Shyam, S. D. Pandey and M.
Dayarani. National Research Centre on Banana No. 44, Ramalinga Nagar South Extn.
Vayalur Road, Trichy 620 017. Pg. 10
2. Seminar on technological advancement in Banana production handling and processing
management - Souvenir 27-28 March 1999 B.M.C. Reddy and Prakash Patil. AICRP on
Tropical Fruits. Indian Institute of Horticultural Research. Hessaraghatta, Bangalore 560 089.
Pg. 1-3,6

31
Climatic Requirements
Banana is strictly a tropical plant, however, it is also grown in subtropical to
humid climates. The optimum temperature required for bananas is between 25
to 30°C, with upper and lower limits of about 40° and lO^C respectively. For
the optimum growth of the banana plant, the area should have a monthly
rainfall of 20-22 mm. distributed evenly. Strom and strong wind of 40 to 60
km/hr can cause damage to leaves and wind velocity of 95km/hr can cause
complete destruction of banana plantations. Banana must be protected from
strong winds by planting windbreak of tall trees in the windward side. 3

Soil Requirement
Banana is grown successfully on a wide range soil. It can be grown on all type
of soils except deep black cotton and pure sandy soils where small remedial
measures are to be taken. Since banana crop is sensitive to water logging, soil
with good drainage with low water table meter should be selected. A soil pH of
5.5 to 8.0 is found to be the optimum.

Varieties
In Maharashtra different banana varieties like Safed Velchi, Dwarf Cavendish,
Robusta, Grand nain, Sindhurni, Hanuman, Ardhapuri, Lalvelchi and Rajeli are
grown. 4

3. Seminar on technological advancement in Banana production handling and processing

management - Souvenir 27-28 March 1999 B.M.C. Reddy and Prakash Patil. AICRP on
Tropical Fruits, hidian Institute of Horticultural Research. Hessaraghatta, Bangalore 560 089.
Pg. 1-3,6
4. Seminar on technological advancement in Banana production handling and processing
management - Souvenir 27-28 March 1999 Table 4 Banana Cultivars grown in different
states. Singh and S. Uma, 1996. Pg. 15

32
Planting
For Dwarf Cavendish varieties optimum plantation i.e. higher density
plantation of (4444 plants per hector) with closer spacing of (1.5 x 1.5 m.) was
recommended. 5 Farmers used the recommended spacing which was calculated
as 5'x5' and plantation of 1700 banana plants per acre.

Fertilizers
The fertilizer recommendation for Cavendish varieties is 200 g. N, 100 g. P 2O5
and 200 g. K2O/PIant/crop. Nitrogen and Potash are to be applied in four equal
split doses at 30,75, 120 and 165 days after planting while that of phosphorus
cab be applied at the time of planting, g

Drip Irrigation
Drip irrigation is network of pipes through which water is supplied to the plant.
Drip irrigation was used for banana plantation which was also useful in
providing liquid nutrients and showed better performance.

Fruiting and Covering

Banana bunches matures in about 110 to 130 days after flowering. The grown
bunches are covered with polythene bags to protect it from the sun.

Yield
The average yield range of Cavendish is 50 - 100 tons / hector, however 150
tons/hector can be obtained under good cultural practices with high density
planting. 7

5., 6., 7. Seminar on technological advancement in Banana production handling and

processing management - Souvenir 27-28 March 1999 B.M.C. Reddy and Prakash Patii.
AICRP on Tropical Fruits. Indian Institute of Horticultural Research. Hessaraghatta,
Bangalore 560 089. Pg. 1-3,6

33
3.2 Commercial Banana Cultivation

Commercial banana cultivation requires specific climatic conditions, soil,

irrigation, fertigation, planting material and post harvest technologies. By
following scientific method of plantation, the grower can produce optimum and
best qualities of yields and earn more profits.

Banana is a crop which can be grown throughout the year. This has least
gestation period where the first yield can be expected in just 11 to 12 months
from the period of transplanting. Reduced gestation period greatly helps in
achieving higher profits. To achieve reduced gestation period, selection of
proper planting material is required.

Planting Material

Banana is commonly propagated through sword suckers having narrow leaves.

Three month's old suckers with developed rhizomes are used for plantation.
Disease free planting material is made ready for plantation. Though some
farmers are using sword suckers for banana plantation, many farmers are now
using Micropropagated plants for plantation. Tissue cultured banana plants are
getting commercial acceptance due to their qualities viz. superior, disease free
varieties and mass production. From tissue culture of bananas no genetic
improvement was expected, yet these plants have proved better, compared to
traditional planting material, only under a good management system.

34
3.3 Difference between conventional and scientific Banana
Farming

Conventional Farming: Suckers are used as planting material, which may

not be free from any infections and can be of inferior quality. In such case,
there can be some mortality. As characteristics of the parent material is not
exactly known, yield upto the expectations may not be achieved. The banana
plant yields, on an average, about 25 kg per plant. Total plant population is
from 1200 to 1380 plants per acre. Water is fed either through flood, furrow or
basin system so there is possibility of under or over water resulting yield
variation. As solid fertilizers are applied manually, there is high labour cost and
the fertilizer is not utilized fully due to either under or over watering. As
flowering and fruiting is inconsistent and at indifferent times, market
acceptability is moderate. Total profit is not so high in unit area, which is due
to less plant population and mortality.

Scientific Farming: Tissue cultured plants are used as planting material,

which are free from all infections, resulting no mortality of plants. The yields
are very high, upto the expectation. Plants yield on an average of about 40 kg
per plant. Total Plant population is about 2003 plants per acre according to
high-density method of plantation. As water is fed through drip irrigation, the
desired amount of water is available directly to the root zone. So higher yield is
expected. As water solubleorliquid fertilizers are used through drip irrigation,
there is a saving of labour cost and fertilizer is used fully as it is directly
available to the root zone. As flowering and fruiting is uniform and timely,
market acceptability is very high. Total profit is very high due to high-density
i.e. high population and nil or negligible mortality.

35
3.4 Brief history of tissue culture of banana

Every plant cell has the potential to regenerate into a single plant. When this
regeneration of the plant cell into a single plant rapidly takes place at optimum
level, it is called as Micro propagation. When a cell is born, it divides, grows or
dies. These cells are kept in a disease free, clean and controlled environment
with artificial light and temperature where cells start growing and give
desirable results.

Soon after, reports of banana plantlets produced by in vitro shoot tip culture
came from Taiwan (Ma and Shii, 1974). Nearer home, intense work was
initiated at the IIHR by Dr. R. Doreswamy and co-workers (Doreswamy et al.,
1983). Eversince, several investigators the world over have reported success
(Cronauer and Krikorian, 1986). g

It was found that with the help of tissue culture technology constant
improvement in the productivity, profitability, stability and sustainability of the
banana plantation system could be achieved.

3.5 Banana Tissue Culture Technology

Scientists experimented on the different nutrient medias and observed where

the plant showed best results. These results were recorded and when
standardized, these medias were called as the Protocols to produce those
particular plants. The media for each and every stage of growth was different.
For Initiation, Muhiplication, Rooting different medias were prepared.

8. Tissue culture propagation of Banana. Dr. R. M. Pandey, Director, Indian Institute of

Horticultural Research. Hessaraghatta Lake P.O. Bangalore 560 089 Pg. 2

36
Banana tissue culture involved following steps:

Procurement of mother plants

Disease free banana plantation areas are located. From these areas, the best
yielding farms are chosen. The elite mother plants are selected from the disease
free farms and maintained under hygienic conditions by spraying fungicides,
bactericides and insecticides. At the time of use, these plants are sterilized and
then used for initiation. In the laboratory five major steps are involved in the
production process.

Stage I : Initiation

In the laboratory, after the surface sterilization of the plant part, the innermost
tissue (ex-plant) is dissected in sterile conditions and put onto the initiation
medium for growth. Initiation medium contains micro and macro elements,
vitamins, irons and growth promoting hormones, solidified by agar agar.
(Figure 1.)

Stage II : Multiplication

This is the next stage to multiply the plants in sterile conditions. When the
tissue starts growing and forms a shoot, it is transferred to another medium
containing growth-promoting hormones that enhance the cell division. The
growing shoot multiplies and forms a cluster of three or four shoots. Same
cycles are repeated for ten to twelve times to reach to the optimum production.
(Figure 2.)

37
stage III : Shooting and rooting

When the plant is ready, it has to be transferred to the rooting medium. The
single shoots are separated and placed onto a shooting and rooting medium. At
this stage the hormones may or may not be required. The shoot elongates and
new roots come up. Rooting takes about three or four weeks and the plant
becomes ready for hardening. (Figure 3.)

Stage IV : Semi-Hardening

In the semi-hardening process, the plants are made ready to sustain in the
natural farm conditions. Hardening is done in the controlled conditions of the
green house. The plants are taken out of the bottle and the media adhering to
the root system is washed fiilly. Afterwards, the plants are graded as per their
size and then transferred singly into the seed tray containing sterile, soil-less
medium (a mixture of peat moss, soilrite, sand or perlite). These trays are kept
in the humidity chambers for six weeks and thereafter they are kept in open in
the Green House. Regular spraying of fungicides, bactericides and insecticides
is done to achieve good hygienic condition of the plants. (Figure 4.)

Stage V : Hardening

In the hardening process, the plants from the seed tray are separated and
transferred into polythene bags preferably black coloured, containing a mixture
of sand, soil, peat moss, soilrite, perlite or compost. These plants are kept in the
shade-house where Fifty to Seventy-five percent of the sunlight is reduced
through the nets and entry of insects is also eliminated. Irrigation is done by
drip in each polybag and sprinklers or misters maintain humidity. This
hardening also takes another six weeks and the plants get fully acclimatized to
environmental stresses. These plants are directly used for planfing into the
field. (Figure 5.)

38
 Figure 1.
Laboratory technicians at initiation stage of banana

Figure 2.
Laboratory technicians at banana multiplication

39
 Figure 3.
Grown banana plants at rooting stage

Figure 4.
Semi-hardened banana plants in humidity chambers

40
 Figure 5.
Hardened banana plant ready for sale

Flow Chart No. 2

Process Flow Chart For Tissue Culture of Banana

Selection of Elite Plants

ight Sword Sucker

Selection of Daughter
i
Surface Sterilization of Sucker and dissection of the shoot
tips under aseptic condition
i
Inoculation of shoot tip on nutrient media
and incubation in controlled microclimatic conditions
i
Establishment of cultures and origin of new shoots
I
Multiplication of Shoots

Elongation or Shooting / Rooting

Primary hardening or rooted plants in green house

Planting in the field

41
Recent experiments conducted to evaluate the performance of tissue
cultured seedlings v/s normal suckers planting in Basrai Banana: 9

A study was conducted to evaluate the performance of tissue culture seedlings

V/s normal suckers planting during 1994-95 at Banana Research Station,
Jalgaon, Maharashtra. The results of this experiments revealed that tissue
culture seedlings planting was found significantly superior over normal sucker
planting for all growth, yield and yield attributing characters studied.

The commercial method of propagation of banana is by rhizome planting. Now

a days, use of tissue culture seedlings as a planting material in banana is
becoming popular in this region. To evaluate the prospects and performance of
tissue cultured seedlings in comparison with normal suckers as planting
material, a comprehensive study was made.

Materials and methods:

The experiment was conducted about materials and methods used in banana
tissue culture during 1994-95. Two treatments viz., use of tissue culture
seedling and normal sucker as planting material in randomized and block
design replicated six times. Almost uniform suckers of Basrai and tissue culture
seedlings were planted at the spacing of 1.5 m x 1.5 m and all plant received
uniform dose of NPK. Observations on plant characters; crop, duration, yield
and yield attributes were recorded.

9. Seminar on technological advancement in Banana production handling and processing

management - Souvenir 27-28 March 1999 CD. Badgujar, S. S. Deshmukh, R. M. Birhade,
C. R. Palwe and N. M. Patil. Banana Research Station, M.P.K.V., Jalgaon -425001. Pg. 23

42
The results obtained in this experimented were as follows:

The observations on growth and yield parameters were recorded and presented.
From the data it revealed that, all the growth and yield characters were found
significantly superior in tissue culture planting over normal sucker planting.
Average height and girth of pseudostem and average number of leaves per
plant 157.35 cm, 63.28 cm. and 36.48 respectively were noticed with tissue
culture plant treatments, which was significantly superior over normal sucker
planting treatment. Least days for flowering (343.00) and harvesting (438)
were noticed with tissue culture planting treatment, which was superior over
normal sucker planting treatment.

The corresponding figures were 380.13, 477 respectively under normal sucker
planting. Thus tissue culture seedlings flowered 37 and matured 39 days earlier
than normal sucker planting.

The yield and yield parameters viz., average number of hands, fingers per
bunch, length and girth of fingers were found significantly superior in tissue
culture seedling treatment. (8.59, 142.07, 2034 cm and 11.46 cm) respectively.
The corresponding figures in normal sucker planting treatments were 6.55,
91.60, 19.60 cm and 10.89 cm.

Maximum average bunch weight (12.69 kg) was noticed under tissue culture
planting which was significantly superior over normal sucker planting
treatment (9.66 kg).

43