Micro Prorogation - Tissue Culture

Introduction:-
Tissue culture, an important area of biotechnology can be use to
improve the productivity of planting material through enhanced
availability of identified planting stock with desired traits. The idea of cell
and tissue culture were put forth by a German Scientist Haberlandt in
1902. All attempts to culture the plant cells and tissue were unsuccessful
till the 1930’s. Around 1939 the possibility of culturing plant
tissues for definite periods was independently reported by white,
Nobecourt and Gautheret.

Micro propagation is one of the important contribution of Plant

Tissue Culture to commercial plant propagation and has vast significance.
The name micro propagation derives Item the miniature shoots/plantlets
initially produced horn this method of plant propagation. Micro
propagation is the true to type propagation of selected genotype using in
vitro culture technique. This technique provides a rapid reliable system for
a production of large number of genetically uniform disease free plantlets
The pioneering work in this area of research was reported by Indian
scientists. Today almost all the prime universities and institutes like BARC,
Bombay, NCL, Pune, ISC Bangalore etc. in India are carrying out research
in tissue culture.

Commercial tissue culture born in India in 1987 when NV. Thomas

& Go. in kerala established their commercial unit for large scale
production of cardarnurn. This was based on the bench scale protocol
developed by Dr. R. S. Nadgauda and Dr. A. F. Mascarenhas at N.C.L.
Pune, India. These pioneering efforts of AVI, Kerala, were followed by
servile other commercial companies who entered in this area. The second
unit was put by Indo-American Hybrid Seed Company at Bangalore in
1988. Now there are several companies in this field viz.

Hindustan Lever, Tata Tea, Unicorn Biotech, Nath seeds, RPG

Enterprises, Indian tobacco, and Hindustan Agri Genetics limited etc. are
the major companies in the field or micro propagation in the country.

There has been an increasing awareness about Bio-technology in

India in recent years. The discussions are not only confined to workshops,
seminars, conferences, symposium but people from various section of the
society are curious F. know and use this technology in various fields.
Modem bio technology offers unlimited opportunities for enhancing genetic
potential of crops, management of biotic and biotic stresses, waste
management by organic recycling. and also faster multiplication of plaiting
material. the technique of bio technology like tissue culture has attracted
the attention of farmers in the Country.

Tissue culture has helped to develop new strain of food crops,

cereals, vegetables flowers, oilseeds and plantation crops such as spices,
coffee, tea and rubber. Maharashtra is in forefront in the use of bio
-technology in the country with tissue culture technology the production of
quality planting material is gaining immense popularity for micro
propagation of various cash crops flowering shrub and trees. Since the
farmers are attracted immensely towards this technology, Govt. of
Maharashtra would like to decide the slate policy regarding use of bio
technology in agriculture. In order to take a stock of present scenario of
various aspects pertaining to bio technology Govt. of Maharashtra has
appointed a committee under the chairmanship of Principal Secretary,
Agriculture of Govt. of Maharashtra. One Sub committee is also appointed
under the chairmanship of Director of Horticulture with the experts to
evaluate the present situation arm suggest appropriate interventions to
boost this technology in the sin ~ Members of this committee are listed as

appendix-I

Such committee is to look into the following issues:-

(A) To collect and consolidate the information regarding present
scenario of micro propagation technology and study the utility of
this technology.
(B) To study the potential for increasing agriculture production with
the help of tissue culture and decide strategy for next 5 years
for use of this technology.
(C) To decide strategy for establishment of tissue culture
laboratories along with criteria regarding financial assistance.
(D) To suggest extension plan for use of micro propagated plants by
the farmers.
(E) To suggest quality control measures for micro propagated
plants.
(F) Decide future direction for research and development in the
field of tissue culture considering needs of the state.

Accordingly the sub committee members met and discussed various

issues, four meetings were conducted on 10/6(99, 5/7199, 1911199
— and 30/7/99. Abstract of discussions is included the report.

Present Scenario:-
Plant tissue culture has been promoted by Government of India
since last 2 decades to enhance the production Land availability of disease
free, true to type, quality planting material.

According to the BCIL, (Delhi) report, at the end of 1996 about 76

commercial micro propagation units have registered for the production
with Government of India of which 30 are functional within the Country.
Total 40 million plants annually are produced against the installed capacity
of 110 million plants.

The current progress of the industry is encouraging but the

expected rapid growth has not taken place. The primary reason for such a
slow growth is because of low awareness level amongst the people about
application of this technology. Although within the country the research
groups have put lots of effort in standardizing protocols for several plant
species, the benefits have not been sufficiently demonstrated to the
farmers at the field level Therefore, the technology is not so popular at
the grass root level.

At the national level, Department of Bio-technology has promoted

the activity with the help of which two pilot plan were set up of the
capacity or 1 million plants per annum of economically important plant
species. Out of these two units one is Ioc8ted ~t National Chemical
Laboratory, Pune, Maharashtra. Extensive efforts have undergone on
developing the experts for setting up of the laboratory and developing
cost effective indigenous technologies with felid evaluation. This unit is
now upgraded to Micro propagation Technology Park to serve the
entrepreneurs/industries institutes/universities so king for mass scale
production of plants.

Recently national workshop on commercialization of plant tissue

culture. Role of Micro propagation Technology park" was convened by
DBT, New Delhi on June 141999 with the major objective of creating an
awareness and present the concept of Micro propagation Technology
Parks.

The potential for the domestic market is enormous and by

conservative estimates it is around RS. 150/- cores with the annual
growth rate of 10%. In order to fulfill the demand of domestic market
even at the meager lever of 10 to 15% efforts needs to concern rate not
only to increase the units at private sector but also the back up at
National and slate level from the government is necessary.

Currently, the focus of the companies is mainly in the floriculture

sector however, micro propagation in banana arid sugarcane is also
gaining popularity.

Recently Bhabha Atomic Research Centre, Mumbai have also done

initial work on micro propagation Cardamom Mulberry and Sandalwood.

Currently, micro propagation technology is available on the shelf for

the crops which can be utilized for field test demonstration and farming.
In Maharashtra a research on Biotechnology is under taken in all four
agriculture universities. In addition Maharashtra State Seeds Corporation
has also established tissue culture laboratory for commercial production of
tissue culture saplings of banana and some ornamental plants. The
information regarding present production capacity, utilization and cost per
plants of tissue culture within the state is given in Appendix-II

There are 17 active public and private sector dealing with

production of tissue cultured planets producing Banana, Sugarcane arid
Ornamentals & Orchids etc. incidentally, Maharashtra is emerging these
crops for shifting their cultivation to such type There are 6 major units in
the State producing plantlet, 7 units producing 15.60 lakh plant around
31.30 lakh plants of ornamental crops particularly gerbera carnation and
orchids. The units in the State are producing 91.85 lakh tissue Cultured
plants every year. This clearly shows that state has already taken lead in
this area recognizing a great scope for this industry in future.
 Maharashtra is a major banana growing state in the country.
Maharashtra tops the list with its higher productivity with area coverage of
about 60000 hectares. In order to increase productivity of banana, it is
highly essential to adopt new production technology to ensure better
quality production. One of the eat that is being posed in recent past is
that Sigatoka (Cercospara muse) which would demand replacement of this
infected planting material Tissue Culture could be one alternative for
faster replacement of planting material. Experimentation has shown that
there is a significant increase in the yield in banana and yield parameter
litter average number of hands fingers per bunch, length and girth or
fingers due to which there is around 30% increase in the yield of banana.
Sugarcane also yields 10 -15 0/) increase in productivity due to tissue
culture plantlets. Our State occupies 5.2 lakh hectors area under
sugarcane with an average productivity or 85.5 MT/ha. Quality planting
ma that in sugarcane is also concern a matter of in sugarcane cultivation.
Once sugarcane was considered relatively less affected by pests and
diseases, however during past few years, a few of rumor pests earlier
nave now play a major role Threatening its productivity in future.

There are 13 major pests reported to damage sugarcane crops

heavily. A sugar industry in Maharashtra plays a vital role in the rural
economy of the State and has successfully boring out economical and
social reforms in rural Maharashtra. State produces more than .33O/~
sugar produced in the country. There has been, however, declining trend
in the yield for the past few years, major reason being lack of good quality
planting material. Using newly developed cost effective protocol of
sugarcane through micro propagation large number of sugarcane plantlets
are being produced in Maharashtra by various public and private sector
units.
One of the major issues apart from planting material has been that of low
emergence of eye bud, thereby reducing plant population, and ultimately
effecting production of mislabel cane. Tissue Culture technique for
production of good quality planting material ensures an adequate and
disease free plant to get enhanced yields.

Tissue Culture in ornamental Plants constitute a major part of the

total lancing material produced in state out of 31.25 lakh ornamental and
other plantlets, gerbera a alone is a major flower crops produced in the
State.
Despite increasing production of Tissue Cultured plants in the State, the
total plants produced by various units are only 73% of the total capacity
created for production. Considering huge requirement of tile State, it is
quite essential to augment these industries by strengthening existing units
and also creating new infrastructure for further production of Tissue
Culture of various high value crops.

Although, this technology has been adopted by the farmers,

considering area covered, adoption is at a very low level. Banana growers
used this technology on about 800 hectors, whereas spread of sugarcane
tissue cultured plantlets is only on 1 W hectare. One of the constraints for
adoption of this technology has Ken an initial high cost of tissue cultured
plants. The initial cost riot bei~4g affordable to farmers compared to the
cost of traditional planting material, there is slow increase in the demand.
The Tissue Culture Units also fail in reducing production cost of planting
material mainly du2 to heavy initial capital investment. The private sector
has good tissue culture plants, but they are not ready with relevant
standardized package of practices to meet field requirement raising field
crop with tissue cultured plants. Lack of an adequate R&D infrastructure
at unit level makes such type of recommendation impossible to private
units arm an turn, grower has accepted to use tissue cultured plaids with
traditional package of practices. This situation not only leads to wastage
market purchased inputs, but also reduces per rupee net return.

Potential:-
Horticulture is an emerging sector of economic importance in
Maharashtra. Similarly sugarcane is also an important industrial crop of
the state. Floriculture has already established i~ place in economic
development of farmers of Maharashtra. Some awareness has been
created in use of micro propagation plants in the cultivation of banana,
and sugarcane. However, coverage under micro propagation plants is
hardly to the extent of 800 hectors in case of banana and very little in
case of sugarcane. Considering the area cove age of banana and inherent
potential of technology, there is tremendous scope to use micro
propagated plants in banana. Similarly sugarcane also offers scope for use
of micro propagated plantlets on a large scale for enhancement of
productivity and better sugar recovery.

(1) Banana:-
Banana covers 60000 hector area present production of micro
propagated seedlings is 36.15 lakhs in banana covering approximately
800 hectors It is proposed to cover 1000 hectors area the first year and
reach the level of 5000 hector in the 5th year with an addition of 1000 ha
area per annum. This would require 45 lakhs tissue cultures seedlings in
the first year and the state will have to reach a production level of 225
lakhs pantalets in the 5th year. In totality this will and to a spread of
tissue culture plants on an area of 15000 hectors considering the use of
daughter suckers in the ration of 1:3. Thus 25 percent of banana in the
state can be covered under this technology which All help to enhance the
productivity of banana in Maharashtra. Even though it is possible to have
larger coverage of area, high cost of planting material becomes prohibitive
for average farmer. It is thus necessary therefore, to reduce the
production cost so that the average farmer can have a access to this
technology. One of [lie reasons for high production cost is the heavy
capital investment, for which the unit holder has to, pay heavy interest.
This interest can be subsidized and converted ii. to soft loan at the
interest rate of.10%. This difference in the rate of interest may be borne
by the state government. For strengthening the existing units a grant
towards interest subsidy may be given to the units to produce additional
25 percent plants over the existing production capacity if they agree to
reduce the rate by Rs.2/- per plant.

(2) Sugarcane:-
 In sugarcane the strategy for increasing are under tissue culture plants
would be three fold. Initially the nurseries of breeder seed would be
prepared by the factories using plants produced by using meristems from
the mother source garden on 100 hector. which will require 15 lakhs
plantlets © 15000 plants/hector for planting the area. The multiplication
ratio being approximately 1:20, the production of sets would be around
300 lakhs sets sufficient to over an area of about 1200 hectors, under
foundation stage seed production. Further the sets for this foundation
stage plants would be used to produce certified stage seed sets sufficient
to cover the area of 12000 hector at an expected multiplication rate of
1:10. Further the sets produced from such certified area will be
distributed to farmers for growing commercial crop of sugarcane on an
area of about 120000 hectors. This program would be maintained by VSI
through the selected sugar factories.
Instead of producing new plants, the existing tissue culture units will have
to be brought under proposed quality control mechanism to ensure
production of good quality and true to type plantlets and subsequent
enhanced production. However, the production capacity upgradation may
be thought of after the midterm review of the program during third year.

(3) Flowers:-
Existing laboratories are also producing flowering species. Since
number of farmers for ploy house horticulture is increasing annually there
is a scope for various use of tissue culture planting material of flowering
plants and vegetable crops. To augment the growing requirement or
planting material, production capacity of existing tissue culture units
involved in production of tissue culture seedlings of flowering species like
Gerbera, Carnation, Lilly, arcadias etc. will have to be increased. With the
existing production of (31 30 laths flowering plants, hardly an area of 50
hect is covered V the green house. Considering the increasing number or
green houses in the state there is a scope of expansion of unites for
production of floriculture plants. It is proposed to reach a level of 100
hectors iii the next 5 years for which production of 65 lakhs plants is
estimated. There are many small scale floriculture units already
established in Maharashtra. Most of the units are importing planting
material horn abroad which consumes very heavy foreign exchequer.
Indigenous production of such Plants would help reducing the burden on
foreign exchequer and also help to reduce capital investment on large
scale iii floriculture units.

(4) Other Potential Crops:-

There are some species of economic importance grown in the state,
turmeric, ginger are the crops where produce has already been developed
in the state. Micro propagation of elite varieties of these crops can be
taken up in the tissue culture laboratories to benefit the farmers.
Propagation of medicinal plants like Saved Musali Asparagus through
tissue culture is also possible. To enhance the utility of infrastructure
established for tissue culture laboratories can be diversified to range of
crops so that units also run in profit.

Strategy for establishment of Tissue Culture

Laboratories
In order to meet the projected requirements. the state government will
have to
(a) Strengthen the existing units arid
(b) Encourage & establishment of new tissue culture units.

(a) Strengthen the existing units

On an average the existing units are running with 75 percent
capacity. The production cost of tissue culture plants is very high and
ranges from Ps. 5.5 to 16 depending upon the crop species. Obviously
such a high cost becomes prohibitive for an average farmer. It is thus
necessary therefore to reduce the production cost so that the average
farmer can have a free access to this technology. One of the reasons for
high production cost is heavy capital investment for which the unit holder
has to pay heavy interest. This interest cap be subsidized and converted
in to soft loan at the interest rate of 10.This difference in the rate of
interest may be borne by the state government for a period of next three
to five years.

For strengthening the existing units a grant ray be given to the

units so as to produce additional 25 percent plants over the existing
production capacity at a rate lower by at least 2/- per plant. This
assistance could be in the form of interest subsidy on the present interest
burden.

(b) Encourage & Establishment of new tissue culture units

Establishment of a new tissue culture units will be treated at par
with other industrial units so that inherent benefit of the industry will
automatically flow to such units also. The zoning declared for the general
industries should also be made applicable to tissue culture units with little
modification treating, all zones as D± Zones. Since the pressure on
industries department is very heavy for giving incentives to different small
scale industries, 100% requirement of incentive amount should be made
available for tissue culture units and should be placed at the disposal of
agriculture department for next five years. In addition to this, there
should be space reservation in the industrial area for tissue culture
laboratories for at least a period of five year.

In order to make these units viable certain policy decision need to

be taken up by the State Government-
i) Tissue culture plants are charged Sales Tax. Although the producer
has to pay the tax, the ultimate burden o~ this tax is shifted on
the farmer. it is proposed that the sales tax on tissue culture
plantlets should be waved at least for initial 10 years.
ii) The electricity tar ii r needs to be brought at par with tariff for
agriculture purpose. For this specific order from government needs
to be issued with an express mention that tissue culture is an
agricultural activity.
iii) The tissue culture units extending hardening facilities in captive
areas, should be given an incentive for polenta paining. Sheds at
the rate or 50 percent of erection cost.
 iv) For establishment of new unit in private sector, grant of Rs. 10
lakhs may be given for the production capacity o~ minimum 5
lakhs plant per annum.
v) The job it venture may be encouraged with the help of agriculture
universities, on the governments lands like TSF,TCDs, and
university farms
vi) 100 percent assistance may be given to the unit holder for
preparation of a project report production capacity.
vii) Wide publicity for such incentives from government may be given
in the state so that more member of entrepreneurs can avail
benefit of this opportunity.

Extension Plan
The extension system today is partially successful in creating
general awareness among farmers about benefits of tissue culture
However, farmers are not clear about me entire package of tissue culture
technology. The presumption that tissue cultured seedlings itself will
increase the yield needs to be addressed in effective manner. Entire
package of practices required for getting optimum yield needs to be
demonstrated effectively on the farmers fields In order to spread the
proper message of benefits of micro propagation, extension machinery
itself needs to be equipped and trained in this area. The entire extension
program for bio technology has to be organized in such a manner that
extension workers and the farmers get simultaneous training and could
upgrade their skills for quality production from tissue culture plantlets.
Agronomic practices for tissue culture plantlets have to be finalized
through research and then have to tb propagated through extension
machinery of the state.

Training
Intensive training program for extension staff may be organized
first to train the master trainers and generate in louse resource persons.

This training may be organized for identified persons in the

institutes of repute. This training should be a season long training so that
the ‘technology per-se and it’s on farm application could be
simultaneously observed and studied by the trainee. In second phase this
trained manpower or resource persons will train the field functionaries so
that the field level workers will also be able to develop technology. During
the first year, there has to be a major thrust this training of staff.

The farmers also need training in this aspects of bio technologies.

i) It is experienced that the farmers who use tissue culture plants is

may it be open land cultivation or the green house cultivation, they
are not fully aware of the crop agronomy in a changed situation.
Their training needs to be concentrated on the production aspects
particularly the agronomy and nutritional management. Agriculture
univer5ti~s may not be able to train such huge number of farmers.
Private se 4cr also needs to be involved in the process of farmers
training. Training farmers should be mandatory on the part of
entrepreneur for which 10 percent financial assistance on the
 amount of effective sale may be given as assistance from the
Government. This training will also help hi generating demand for
tissue culture plants.
ii) Sugar industry should also be actively involved in tissue culture
technology. Sugar factories may be persuaded U establish small
tissue culture laboratories in their operational area VSI can take
lead and coordinate these activities.
iii) There needs to be a regular interaction arid continuous dialogue
between the develop in en t agencies, extension workers and
Regular work shops, seminars, symposium needs to be organized
for making farmers aware and also getting feed back from them.
electronic media will play a vital role in propagating this technology
There is a need to produce video films and CDs detailing the
production technology along with success story.
iv) Technology Demonstrations
Through there is an awareness about use of tissue culture in
agriculture there is hardly a demonstration on tissue culture
technology exhibiting total package of practices .Extension
machinery and the private sector will have to be aggressive in
organizing demonstration on the farmers fields .Listing out the
technology generated in various crops will be the first stage in this
process. Identification of user farmers will be the second stage.
There should be a cross section of farming community
representing the different tissue culture trails. The demonstration
should be minimum 0.2 hector area so that impact of the
technology will be visible.
The assistance for organizing demonstrations should be given by
the state government and by respective private sector. There should be a
fan mer rally on the demonstrations plots to acquaint farmers with the
result of this technology. Farmers trips needs to be organized from the
areas where technology adoption is low to the areas with high level of
technology adoption may it be in the state or out of the stale.

Quality Control Mechanism

Although tissue culture laboratories have established in the state
during last decade producing tissue culture o plants for distribution to the
farmers, there is no mechanism to checked the quality of planting
material produced by them. Since the quantum of tissue culture plant
would be increasing at higher rate The quality of plants will have to be
ensured for getting desired type of quality planting material. It should be
mandatory for all the tissue culture units to register themselves with the
competent authority designated by the state 90\ eminent. This will help
monitoring the production and also the quality of sources of multiplication.
The plants produced through tissue culture technology will also be
subjected to the provisions under existing Seed laws, Rules and Orders
Present enforcement agency of the state government. would be deployed
to undertake regular inspections and checks to ensure supply of quality
planting material. The purity standards of a particular clone/variety should
be ascertained by raising the mother clones in the genetic garden which
will be maintained by the state agriculture universities aid institutes like
VSI. There should be a mechanism of regular inspection of laboratories by
a team constituted by the Government. This team will fix the cities for the
proper working of laboratory, sanitation of [lie equipment, (All production
related infrastructure and undertake regular ii inspections of the
production units.

One central quality testing labor Atari government in public sector.

Those LII laboratory will avail this facilities by pay as fixed by state
government. One laboratory should be established by the to test the
samples drawn by info amendment should be made in the exit will
established by state its without the quality testing lug app operate service
charges central Fever quality testing qovemnierit under public sector
icei’iient autlmrity. Appropriate sting seed laws to incorporate
provisions for quality control of tissue culture.

Future Research and developed needs

At present the cost of tissue culture planted are quite high due to
how multiplication rates in many protocol efforts need to take place in this
field for example, banana plants multiplication needs to be taken from 2.5
- 3.0 at last 5.00 Various issues regarding decrease in cost needs to be
concentrated for at least next five years. Secondly agronomic practices
commensurable with tissue culture technology have not been finalized so
far. This need to be taken up by agricultural universities and Vasantdada
Sugar institute with strategy to propagate it amongst farmer in nest five
year. Thirdly new protocol. need to be firmed up and propagated for these
crops which have tremendous potential for Maharashtra, those may
include medicinal and aromatic plants , spices, mango and cashew nut for
state ’s agro climatic conditions. Fourthly, be spelled out and
quality testing facilities standardization of such quality parameters so that
provision of seed Act, Rules and Seed Control Order could be effectively
enforced is a pre-requisite for quality control measures, Fifth import
replacement substitutes need to be developed so that quality outflow of
royally could be eliminated in may of the flower varieties. If would also
save precious for exchange . Finally extension methodologies and post
harvest measures need to be addressed so that tissue culture technology
expands and quality product reach up to the consumer end of good
economic returns .

TISSUE CULTURE CENTRES

1. Central Research Institute for Dryland Agriculture

Santoshnagar, Hyderabad - 500059
Tel :4530177, Fax:4531802

2. Chowgule Industries Ltd.

No. 2. 2nd Floor, Unity Bldg,
Tower Block. Mission Road,
Bangalore - 550002.
Tel : 2224171 / 2235583
Fax :2226217

3. Cornpackt
16-A, South End Road,
Basavanagudi
 Bangalore - 560004.
Tel : 6647847
Fax : 6647637

4. Growrnore Biotech (P) Ltd.

418, Sipcot Phase 2, Moranapalli
Village Hosur (East Street)
Bangalore -635109.
Tel : 25565 Fax: 25565

5. Indo American Hybrid Seeds (lndia) Pvt. Ltd.

2nd Main, 17th Cross, Kr Road,
BSK 2nd Stage
Bangalore – 560070
Tel : 6650111 Fax : 6650479

6. Khoday Biotech
Palm Groves Nursaries, 7th Mile.
Karnataka Road
Bangalore – 560062
Tel: 6641886 / 8432441
Fax : 6644680

7. Kurnar Gentech & Tissuecullures Go

Kumar Capilal
2413 East Steet, Gamp
Pune -411001
Tel: 650065 / 672052
Fax: 653365

8. Laxmi. Associates
4/21, Marg Darshan
Prof. N. S. Phndke Marg,
Andheri ((B)
Muinbai - 4000069 t
Tel :8373538
Fax :8373538

9. Somniya Organo Chemicals Ltd.

4th Floor, Tower Block.
Unity Building, J. C. Road,
Bangalore - 560002.
Tel: 2236479 / 219103
Fax: 2219103

10. Varsha Agro Industries

61/C, Ghikkallasandra
Subramayapura Posi
Bsngalore – 560061
Tel: 6699591 /6593394
Fax : 5693394