chapter no. 2 introduction to plant tissue culture

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Chapter No. 2 Introduction to Plant Tissue Culture

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Page 1: Chapter No. 2 Introduction to Plant Tissue Culture

Chapter No. 2

Introduction to Plant Tissue Culture

Page 2: Chapter No. 2 Introduction to Plant Tissue Culture

INTRODUCTION TO PLANT TISSUE CULTURE

Contents

2.1 Introduction and Definition

2.2 Advantages of Plant Tissue Culture

2.3 Objective of Tissue Culture Project

2.4 Conceptual Foundation of Plant Tissue Culture

Technology

2.5 History of Plant Tissue Culture

2.6 Applications of Plant Tissue Culture

2.7 Flow Chart of Plant Propagation by Tissue Culture

IVIethod

2.8 Important Steps in Plant Tissue Culture

2.9 Commercialization of Plant Tissue Culture Technique

2.10 Indian Scenario

2.11 Assistance for Plant Tissue Culture Business from

Different Sources

2.11.1 Assistance from Government Agencies

2.11.2 Assistance from Micro Propagation

Technology Parks

2.11.3 Assistance from Financial Institutes

2.11.4 Assistance for Marketing

2.11.5 Assistance from Biotech Consortium India

Limited

2.12 Conclusion

Page 3: Chapter No. 2 Introduction to Plant Tissue Culture

INTRODUCTION TO PLANT TISSUE CULTURE

2.1 Introduction and Definition

Agriculture today is on the verge of a technological revolution, in a manner it

has never been seen before. As we enter the new millennium, one key

development that comes to our mind is the emergence of biotechnology, which

offers some of the best opportunities and solutions to some of the

uncontrollable problems faced by us.

Biotechnology is a group of technologies that share two things in common;

they manipulate living cells and their molecules and have a wide range of uses

that can improve our lives. The major techniques of biotechnology are genetic

engineering, cell culture, tissue culture, bioprocessing, protein engineering etc.

Plant Tissue Culture, Cell Culture or Micropropagation is the technique of

producing selected plants of known desirable agriculture qualities, in large

numbers of plants from small pieces of plant in relatively short periods of time.

It is a method of rapid propagation under controlled disease free conditions.

Entire crop population with premium qualities can be created from a single

elite specimen plant. Depending on the species in question, the original tissue

piece may be taken from shoot tip, leaf, lateral bud, stem or root tissue of the

mother plant. Ex-plants from selected mother plant are established and

multiplied under 'In-Vitro' conditions, providing the optimum pre-requisite for

plant growth. These ex-plants go through the initiation, multiplication and

rooting methods for producing a cell into a full-fledged plant. These ready

plantlets are then hardened in climatically controlled green houses or poly

houses. Depending on the species, the plants become ready for plantation in the

field. This technique of plant propagation greatly reduces the labour and space

Page 4: Chapter No. 2 Introduction to Plant Tissue Culture

requirement, for producing new varieties and can also markedly enhance

propagation rates.

Mettiods of plant propagation and establishment are of particular interest to our

country and work on a wide range of vegetables, fruit crops and trees are in

progress. Several scientists have been experimenting to extend the application

of tissue culture to make plant species commercially important. For example

experiments on crops like coconut, date palm. Cashew, Mango, Orange etc. are

being made in the different research laboratories.

With the advent of plant tissue technology, it is now possible to propagate fine

varieties of flowers, forest, and fruit trees by tiny plantlets. Commercialization

of these crops has already taken place. In Ornamental crops, Orchids,

Carnation, Gladiolus, Gerbera, Anthurium etc. has been commercially grown.

In Fruit crops, Banana, Sugarcane etc. has been commercially grown. In Forest

trees. Teak, Eucalyptus etc. has been commercially grown. Medicinal plants are

also being experimented by tissue culture method and soon would be ready for

commercial plantation.

Biotechnology is an area with a tremendous potential in solving basic problems

of food, fiber, fuel and medicine particularly in developing Asian countries.

2.2 Advantages of Plant Tissue Culture are as follows

• Mass multiplication of elite clones: Micropropagation allows the

production of large numbers of plants from small pieces of the mother

plant. The production requires relatively short periods of time to grow

plants. Depending on the species under production, a single ex-plant can be

multiplied into several thousand plants in less than one year.

Page 5: Chapter No. 2 Introduction to Plant Tissue Culture

<• Elimination of diseases in planting material: Another purpose for which

plant tissue culture is uniquely suited is in the obtaining, maintaining, and

mass propagating of specific disease-free plants. The concept behind

indexing plants free of pests is closely allied to the concept of using tissue

culture as a selection system. Plant tissues known to be free of the disease

under consideration (viral, bacterial or fungal) are physically selected as the

explants for tissue culture. Tissue culture could be a useful way of

circumventing or eliminating disease, which can accrue in stock plants.

•J* Plant improvement through tissue culture: Creation of superior varieties

of agricultural crops is possible through tissue culture method, which

otherwise is not possible through conventional plant breeding methods.

•> True to Type production: Large number of true to the type plants could be

propagated within a short time and space and that too throughout the year.

For example, it may be possible to propagate Two to Four lakhs of tissue

cultured plants from a single bush or rose against 10 to 15 plants by

conventional means. Also, it may take about Two to Four months to

produce a healthy planting material by tissue culture means, whereas a

minimum of Six to Eight months is required for most species by the latest

method of plant propagation.

•> Higher Yields: Tissue Culture Plants may have increased branching and

flowering, greater vigour and higher yield, mainly due to possibility of

elimination of diseases.

<• Beneficial when conventional propagation is difficult: The method may

succeed to propagate plants where seeds or conventional propagation is not

possible or difficuk or undesirable.

• Efficient method in saving space and energy: The method saves space

and energy of the farmer. For example, in a conventional method the plants

are grown in the open farm requiring an area of about 25,000 m , same

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Page 6: Chapter No. 2 Introduction to Plant Tissue Culture

number of plants would require only 10 m^ space, if they are grown in the

tissue culture laboratory.

•> Flexible method: The flexibility of nurseries can be improved. As the

capital investment on mother plant is reduced to almost zero, it may be

easier to adapt to changing conditions. Additionally, a better programme of

production is possible, because of the greater plant uniformity and the

availability in the mass at any time.

• Innovation of new varieties: Tissue culture can be utilized for breeding

new varieties.

2.3 Objective of Tissue Culture Project

The primary objective of tissue culture project could be propagation of large

quantity of good quality planting material from elite mother plants within short

time, space with minimum cost per plant.

2.4 Conceptual Foundation of Plant Tissue Culture Technology i

Plant tissue culture refers to the cultivation "In-Vitro" (Vitro-Glass) of all plant

parts, whether a single cell, a tissue or organ, under disease-free conditions on

nutrient medium.

In the life cycle of any organism, two gamates of opposite sex fuse to form a

single cell-zygote. From this single-celled zygote originates the entire

multicellular and multiorganed body of a higher organism. In a flowering plant,

for example, structures as functionally diverse as underground roots, green

leaves and flowers all arise from the single-celled zygote through millions of

divisions of cells.

1. Plant Tissue Culture Course Material, Modern College

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Page 7: Chapter No. 2 Introduction to Plant Tissue Culture

Theoretically, therefore, all the cells in a plant body, whether residing in the

flowers, conducting tissues or root tips, should have received the same

genetical material as originally present in the zygote. There must be some other

factors superimposed on the genetic characteristics of cells, which bring about

this vast variation expressed by the genetically identical cells. The process of

variations is called differentiation. This differentiation is actually preceded by

certain cellular and sub cellular changes. A question that arises at this stage is

whether the cellular changes underlying differentiation of various types of cells

are permanent and irreversible or whether there is merely an adaptive change to

suit the functional need of organism in general.

During the normal life cycle of a plant, it is believed, that the events leading to

differentiations are of permanent nature. However, the experiments of

Vochting on polarity in cuttings (1878) suggested otherwise. He had observed

that all cells along the length are capable of forming roots as well as shoots but

their density is decided by their relative positioning of the cutting.

The best way to answer this question and understand more about the

interrelationship between different cells of an organ and different organs of an

organism would however be to remove them from the influence of their

neighbouring cells and tissue and grow them in isolation on nutrient media.

This has led to the foundation of a new branch of biology as 'Cell and Tissue

Culture'. It is applicable to both plant and animal cells. Plant tissue culture has

acquired many practical applications in agriculture, horticulture and forestry. It

is increasingly becoming popular as a part of recent field of Biotechnology.

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Page 8: Chapter No. 2 Introduction to Plant Tissue Culture

2.5 History of Plant Tissue Culture 2

The German Botanist Guttlieb Haberlandt first proposed the importance of

plant tissue and cell culture in isolation, in 1902. He is regarded as the father of

plant tissue culture. He used tissue of Lamium Puroureum and Eichhornia

crassipes, the epidemis of Ornithooalium and epidermal hairs of Pulmonaria

Mollissima. He grew them on a particular salt solution with sucrose and

observed obvious growth in the cells. The cells remained alive for up to 1

month. They grew in size, changed shape; thickening of cell walls occurred and

starch appeared in the chloroplasts, which initially lacked it. However, none of

the cells divided. The failure was that he was handling highly differentiated

cells and the present day growth hormones, necessary for inducing division in

mature cells, were not available to him.

Hanning (1940) had initiated a new line of investigation, which later developed

into an important applied area of in-vitro techniques. Hanning excised nearly

mature embryos of some plants like Raphanus Sativus and successfully grew

them to maturity on mineral salts and sugar solution. Van Overbeck (1941) and

co-workers demonstrated for the first time the stimulatory effect of coconut

milk, which was similar to embryo sac fluid, on embryo development and

callus formation in Datura. This proved a turning point in the field of embryo

culture, for it enabled the culture of young embryos which failed to grow on a

mixture of mineral salts, vitamins, amino acids and sugar.

Subsequent detailed work by Raghavan and Torrey (1963), Norstog (1965) and

others led to the development of Synthetic media for the culture of younger

embryos. Laibach (1925, 1929) demonstrated the practical applicafion of

2. Plant Tissue Culture Course Material, Modern College

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Page 9: Chapter No. 2 Introduction to Plant Tissue Culture

embryo culture in the field of plant breeding. He isolated embryos from non­

viable seeds of a particular plant and reared them to maturity on a nutrient

medium.

In 1922, working independently Robbins (USA) and kotte (Germany) reported

some success with growing isolated root tips. White made the first successful

report of continuously growing tomato root tips in 1934.

During 1939 - 1950 extensive work on root culture was undertaken by Street to

understand the role of vitamins implant growth and shoot-root relationship.

Gautheret (1934), White (1939) and Nobecourt successfully cultured cells of

Salix, Nicotiana-Hybrid and carrot on synthetic media. They, for the first time,

demonstrated that growth regulators and vitamins if added to media enhanced

the growth forming mess of cells called callus.

Skoog (1944), Tsui (1951), and Miller (1955) demonstrated the induction of

divisions in isolated, mature and differentiated cells by using synthetic as well

as natural compounds. Muir (1953) developed a technique of growing single

cells into liquid medium in case of Tenetes Erecta and Nicotiana Tabacum.

Vasil and Hildeprandt (1965) raised whole plants starting from single cells of

tobacco. Skoog and Miller (1957) showed that changing the relative

concentrations of the two substances in the medium could regulate the organ

differentiation.

The first reports of some embryo formation from Carrot tissue appeared in

1958-59 by Reinert (Germany) and Steward (USA). Ball (1946) successfully

raised whole plants of Lupinus and Tropaeolum by culturing shoot tips. Morel

and Martin (1952), for the first time, recovered virus-free Dahlia plants from

infected individuals by culturing their shoots.

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Page 10: Chapter No. 2 Introduction to Plant Tissue Culture

Murashige (USA) used this technique to muUiply plants in large number for

several species ranging from ferns to foliage, flower and fruit plants. Guha and

Maheshwari (1966) demonstrated the possibility of raising large numbers of

plantlets from pollen grains of Dhatura. In 1972 Carlson and others produced

the first somatic hybrid between two plants by fiising their protoplasts.

2.6 Applications of Plant Tissue Culture 3

In the early fifties it was observed that plant cells are amenable to chemical

manipulations in the medium whereby they can be induced to form organized

structures and complete plants. This discovery is considered to be very

important for the application of cell and tissue culture methods to overcome

several problems connected with agriculture, horticulture and plant breeding.

1) The technique provides a way for rapid multiplication of desirable and rare

plants. 20,000 plants/year/bud in turmeric, 1,00,000 plants/year/bud in

Eucalyptus were found.

2) As the experiments reveal, virus infected plants also contain some healthy

stocks as such they can be obtained by separating shoot tips for their in-vitro

propagation. This has given successfiil results in Strawberries and Sugarcane.

3) The development of haploids through the technique of anther culture has a

potential significance in basic and applied genetics and plant breeding. During

the past 20 years the technique has been successfully extended to about 20

plant species including some economic plants.

3. Plant Tissue Culture Course Material, Modern College

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Page 11: Chapter No. 2 Introduction to Plant Tissue Culture

4) The embryo culture has been useful in overcoming seed dormancy. It is also

utilized for producing viable plants from crosses, which normally fail due to

the death of immature embryos. Experiments were successftil in case of Jute

and Rice.

5) The embryo tissue culture is also applied for the propagation of rare plants.

In some experiments, coconuts developed soft, solid and fatty tissue in place of

the liquid endosperm (Mohan Ram, 1976). These are rare and very expensive,

served only at special banquets in Philippines. Under normal conditions the

coconut seeds fail to germinate. Using the technique of in-vitro culture of

excised embryos De Guzman (1969) succeeded in making plantlets from

makapuno nuts.

6) Another important use of embryo culture is found in obtaining some rare

hybrids. It is possible to raise complete hybrid plants through embryo culture.

This method has been profitably used for many interspecific crosses of crops

like Tomato, Papaya and Cotton.

7) It is possible to isolate and culture single cells of plants. This helps in mutant

selection in relation to crop improvement, as done in Tobacco, Datura etc. The

technique is also useful in the production of some chemical substances in the

industry. In some cases cell cultures contained twenty times more chemical

content than the roots.

8) Recently tissue culture is used in protoplast culture of different varieties of

plants and these protoplasts are used for somatic hybridization.

9) A few high performance crop varieties have been widely adopted, resulting

in the disappearance of a large number of older varieties. The forests, which

house the wild races of most of the crops, are being cut on large scale. Hence

tissue culture can be used to preserve germplasm i.e. tissue conservation of

these plants can be done identically.

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2.7 Flow chart of plant propagation by tissue culture method

PREPARATION OF MEDIUM

I STERLISATION IN AUTOCLAVE

POURING IN BOTTLES

SEALING AND RESTERILISATION

COOLING AND SETTING

I READY FOR INOCULATION

SUBCULTURING

SELECTION OF A MOTHER PLANT

I CUTTING OUT THE EX-PLANT PART

WASHING EX-PLANT IN WATER, SOAP AND ANTISEPTIC SOLUTION

I REWASHING AND SURFACE STERILISATION OF EXPLANTS IN CHEMICAL SOLUTION

SHIFTING OF EXPLANT TO LAMINAR AIR FLOW STATION

SURFACE STERILISATION IN SODIUM HYPOCHLORITE FOLLOWED BY WASING IN DISTILLED WATER FOR 3-4 TIMES

INOCULATION OF EXPLANT ON STERILISED MEDIUM OF KNOWN COMPOSITION

SHIFTING OF CULTURE TO GROWTH ROOM (I5-25''C, 3000 LUX)

3-6 WEEKS

BUNCH OF IN-VITRO SHOOTS (GROWTH)

SEPARATION OF IN-VITRO SHOOTS

TRANSFER TO ROOTING SOLUTION IN GROWTH ROOM

-3 WEEKS

TRANSFER TO GREEN HOUSE FOR HARDENING

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2.8 Important Steps in Plant Tissue Culture

1. Preparation of medium and sterilization in Autoclave: In the commercial

laboratory, the medium is prepared for initiation, subculture and rooting

purposes. The medium comprises of the nutrients such as Micro Nutrients,

Macro Nutrients, Vitamins, Irons and growth regulators required for the growth

of the plant. The medium is sterilized in the electrically operated Autoclave at

Fifteen pounds temperature. This sterilization is important to avoid the

bacterial contamination. This medium is then poured into the culture bottles

and these bottles are again sterilized in the autoclave. Then the medium is

cooled and is ready for inoculation.

2. Selection of a mother plant and sterilization: The commercial laboratory

decides the plant species, which are to be multiplied in the laboratory.

Accordingly, the mother plants are selected from the virus free areas. Healthy,

disease and virus free plants are selected as mother plants. The actual plant

part, which is called as ex-plant is selected for inoculation. This plant part is

washed with water, liquid soap and antiseptic solution. This ex-plant is washed

again with the chemical solution to avoid any fungal contamination coming

from the field environment. To remove the remnants of the chemicals it is

thoroughly washed by distilled water.

3. Inoculation of explants on sterilized medium of known composition:

Inside the laminar flow station, the ex-plant is treated again with a disinfectant-

Chemical called Sodium Hypochloride. After this treatment the ex-plant is

washed thrice with double distilled water to remove the chemical remnants.

The explants are inoculated on the sterilized medium. The inoculation takes

place in the Inoculation room, on the Laminar Air Flow station, which

maintains a continuous air current, which keeps this bench without the risk of

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contamination. The expert technicians in the laboratory do the inoculation.

(Figure 1, 2)

4. Shifting of cultures to the growth room: Inoculated cultures are then

shifted to the growth room where a typical temperature of Fifteen to Twenty

degree centigrade with the help of air conditioners and Class 1000 clean air is

maintained. The artificial lighting arrangement is also made for the growth of

the plant. (Figure 3)

5. Shifting of cultures for Subculture: After three to six weeks from

inoculation, the inoculated ex-plant shows growth in multiple shoots. These

shoots are transplanted on the subculture or multiplication medium for ftirther

growth of the plants. For subculture, separate growth medium is prepared.

Several subculture cycles are done in the laboratory for mass production from

the ex-plant. (Figure 2)

6. Separation of in-vitro shoots and rooting: The shoots are separated in the

laminar airflow stations and these fully grown shoots are transferred to the

rooting medium for root generation. Depending upon the species, rooting

requires One to Three weeks. (Figure 2)

7. Transfer to green house for hardening: Rooted shoots are removed from

the laboratory and are transferred to the green house for hardening. In the

hardening procedure the plants are first kept in the humidity chambers for

acclimatization and then are transferred to the green house. (Figure 4)

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Figure 1.

Closer look of ex-plant at inoculation

Figure 2.

Laboratory operators at inoculation, subculture and rooting work.

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Page 16: Chapter No. 2 Introduction to Plant Tissue Culture

Figure 3.

Cultures in Growth Room

X V

Figure 4.

Plants transferred to green house for hardening

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2.9 Commercialization of Plant Tissue Culture technique

In Western Europe, commercial micro-propagation started as late as early

1980's and there were total two hundred and forty commercial plant tissue

culture laboratories producing millions of plants per year by the end of 1988.

Even Holland started commercial micro-propagation in 1980's and by the end

of 1988 Holland was producing sixty two million plants with the help of sixty-

seven tissue culture laboratories. Even Israel had five commercial tissue culture

laboratories producing five million plants per year by the end of 1988.4

Also in other countries many commercial tissue culture laboratories were set up

and produced millions of plants for their country. These countries included,

Poland, Yugoslavia, and Soviet Union etc.

In America commercial micro-propagation started in 1965 and there were

about hundred commercial tissue culture laboratories then.

2.10 Indian Scenario

In 1980's while all these countries were producing millions of plants, India had

only four commercial tissue culture laboratories. Eventually the laboratories

increased, but they were unable to produce the quantity that agriculture and

horticulture market needed.

Many commercial plant tissue culture laboratories commenced operations in

1990's. Currently the plant tissue culture is well studies, experimented and

4. Biotechnologies for Agriculture and Aqua Culture. Chapter 27-Present capabilities in Commercial Tissue Culture and the Potential for Growth. Dr. Jitendra Prakash 195-199

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accepted in India. India has achieved a milestone in this technique by

conducting research and development with well-equipped research laboratories

like; Indian Council for Agriculture Research, Delhi; Indian Institute of

Horticulture Research, Bangalore and National Chemical Laboratory, Pune.

Now the need is to make this technique strong in the commercial area of

production.

Despite of the support of the national level Research Institutes, Agricultural

Universities and Government Agriculture Department, commercial tissue

culture is still facing multifarious problems.

2.11 Assistance for Plant Tissue Culture business from different

sources

It was found that different type of financial and technical assistance was

available for commercial plant tissue culture laboratories. The commercial

laboratories did not identify these sources. Following are the different

assistance schemes available for commercial tissue culture laboratories.

2.11.1 Assistance from Government Agencies

Various Central and State Government agencies have been trying hard to boost

agriculture business and agriculture processing sector in the State. The lacking

factor is, a dialogue between the farmers, agriculture industry and the

Government development agencies. To fill up this gap. Government

departments have come up with different schemes. Most of the commercial

laboratories were not aware about these schemes. Commercial laboratories

should study these schemes and make use of them in making their business

cost-effective.

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Schemes implemented by Central and State Government for promotion of

Agri Business and Agro Processing Sector:

1. Name of the scheme: To establish Tissue Culture Laboratory in private

sector.

Scheme implementing agency: Central Government

Nature of Assistance: Assistance is available for one unit per year for

private sector for establishing tissue culture laboratory. Subsidy available is

Rupees Ten Lakhs.

2. Name of the scheme: Assistance for establishing Green Houses.

Scheme implementing agency: Central Government

Nature of Assistance: Under this scheme for establishing Green Houses for

high tech agriculture following assistance is available.

a) G.H.I: Green house frame and U.V. Film would be subsidized upto

Fifty percent with a limit of rupees Thirty-one thousand two

hundred and fifty only for one green house.

b) G.H.2: For partially controlled green houses using Fan and Pad,

subsidy at the rate of forty percent with the limit of rupees One lakh

is available.

Both the schemes of green house are provided with the assistance only for

five hundred sq.mtr. of area for each beneficiary.

3. Name of the scheme: Assistance for Drip Irrigation for High Value Crops.

Scheme implementing agency: Agriculture Department, Government of

Maharashtra

Nature of Assistance: Under Centrally sponsored scheme assistance is

provided for drip irrigation system for fruits, flowers and vegetables. The

rate of subsidy is ninety percent for B.C., S.T., small and marginal farmers

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and women. For other farmers the rate of subsidy is seventy percent.

Subsidy is available for an area upto limits prescribed under Agricultural

Land Ceiling Act. The rate of subsidy is rupees twenty five thousand per

hector as the maximum limit.

4. Name of the scheme: Property tax on Green House

Scheme implementing agency: Government of Maharashtra

Nature of Assistance: Green houses and Poly houses build for high tech

cultivators of Vegetables, Flowers and Nursery plants will not be charged

property tax by village panchayats.

5. Name of the scheme: To provide electricity at a concessional rate for high

tech. agriculture.

Scheme implementing agency: Maharashtra State Electricity Board.

Nature of Assistance: Electricity rate for high tech agriculture like tissue

culture, green houses in Private and Public sector will have concessional

rate as follows:

a) Rupees two and twenty-five paise per unit for high tech agriculture,

which would include tissue culture and mushroom cultivation.

To avail the benefit from Maharashtra State Electricity Board for a

concession in electricity charges, the laboratory should be located out side

the Industrial area.

2.11.2 Assistance from Micro propagation technology parks

In order to promote tissue culture activities, the Department of Biotechnology,

Ministry of Science and Technology, Government of India has developed

Micropropagation Technology Parks. At present there are two

Micropropagation Technology Parks in existence. One is located in

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Maharashtra at the National Chemical Laboratory, Pune and the other is at Tata

Energy Research Institute, New Delhi. The technology parks are developed to

provide an effective platform for transfer of proven technologies to the

entrepreneurs in the field of commercial plant tissue culture. The technology

parks also act as an interface between the Research Institutes and the tissue

culture business for accelerating commercialization of the tissue culture

technology.

Micropropagation technology parks offer following services to the tissue

culture business:

1. Technology transfer: Transfer of proven technologies to the users,

training, implementation and adoption of the technology at the client's

site.

2. Contract research: Development of process for the newer crops,

refinement of existing protocol.

3. Technical assistance for production of plants: An indigenously

designed highly sophisticated laboratory and green house exists to

produce plants on large scale. A strong group of scientists and highly

experienced staff are available with the unit and give technical

assistance for production of plants for plantation programs or field trials.

4. Training of the personnel: Training programs are organized on general

training for plant tissue culture and specialized training on specific plant

of interest.

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5. Setting up and commissioning of tissue culture laboratory and green

house: Designing of the laboratory and green house, equipping the

laboratory and the green house.

6. Advisory consultancy for overall running of the laboratory:

Consultancy provided for running a tissue culture unit, which includes

all the aspects, right from washing of glassware till production and

hardening of plants in the green house.

7. Turn-key project based on the client's need: All the above services

can be taken singly or in combinations as per the needs. Turn-key

project is also offered which includes all the above services together and

any other related aspects or problems.

2.11.3 Assistance from Financial Institutes

Financial assistance is available to tissue culture business houses from the State

Bank of India, Bank of Baroda, Central Bank of India, Dena Bank and

NABARD. The terms and conditions of assistance differ from institution to

institution.

2.11.4 Assistance for Marketing

National Horticulture Board, Ministry of Agriculture, Government of India was

set up in 1984 for integrated development of horticulture in the country.

National horticulture board encourages and promotes the development of

horticulture industry in the country. The board helps in increasing production

and marketing of horticulture produce.

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National Horticulture Board's developmental program in marketing:

Development of marketing of horticulture produces through participation of

Soft Loan.

Objective: To increase flow fresh and processed horticultural produce to

targeted domestic and external markets.

Commodities: Fruits, vegetables of commercial importance

Assistance: The loan support will be made upto forty percent of the loan

portion upto a limit of rupees hundred lakhs per project @ four percent

surcharge per annum, repayable in five installments after a moratorium of three

to five years.

2.11.5 Assistance from Biotech Consortium India Limited

Biotech Consortium India Limited was set up with the objective of providing

the linkages to facilitate accelerated commercialization of biotechnology. This

organization was incorporated as a public limited company in 1990 under the

Indian Companies Act 1956. It is promoted by the Department of

Biotechnology, Government of India.

Biotech Consortium India Limited has been engaged in technology

development, technology transfer, project consultancy, fund syndication,

information dissemination, manpower training and placement related to

biotechnology.

It has assisted over One hundred and twenty clients including scientists,

technologists, research institutions, universities, first generation entrepreneurs,

the corporate sector, government, banks and financial institutions.

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2.12 Conclusion

Propagation by tissue culture offers good commercial prospects in ornamental

plants, vegetables and fruit plants, where value of the product is high. In India

the tissue culture technique has reportedly been successful in more than a

hundred species of plants. It has been estimated that in India, more than Three

hundred and fifty million tissue cultured plants are being produced annually

through tissue culture method.

Plant Tissue Culture has come to stay as a tool in plant biology. Plant Tissue

Culture has the potential to resolve the problems of experimental biology,

which otherwise through conventional methods is difficult to tackle. In the near

future this technique will play a very prominent role in genetic engineering,

breeding and afforestation programs. Tissue culture technique is a boon for the

agriculture and the horticulture industry because of its numerous advantages.

Tissue culture can produce several number of healthy, virus free and true-to-

type plants. The advantage of this technique is such that these plants can be

planted anytime during the year, which solves farmers' seasonal plantation

problems.

Disease-free plants, multiplied through tissue culture, produce higher yields

than infected ones. This biotechnology application can be used for both,

traditional and new varieties. Carefully monitored production would ensure

clean plantlets for distribution.

Tissue culture activity is taking a shape of an industry as many farmers are

planting tissue culture grown plantlets; agro-traders are buying and selling

tissue culture grown plantlets while some are exporting either the plantlets or

the produces of plants grown by tissue culture, especially varieties of flowers

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like roses, anthuriums and gerberas are enjoying high profits. Tissue cukure is

the greatest advancement in plant breeding. Agriculture and horticulture

industry should take full advantage of this technique in reaching the greatest

heights at national and international levels.

In India, tissue culture is rapidly becoming a commercial method for

propagating new and rare species and difficult-to-propagate plants. From a few

research laboratories several years ago, a whole new industry is emerging.

Currently, the demand for micro propagated plants is greater than the supply

with some plants. Some growers specialize in only the micro propagation of

plantlets, leaving the growing-on i.e. hardening activity to others. Many

growers are integrating a tissue culture laboratory into their overall operation.

While many plant tissue culture laboratories are coming up, some of the

laboratories are being closed down due to various reasons and many of the

existing laboratories are found to be complaining about the problems they are

facing. Commercial use of plant tissue culture technique has vast business

potential if tissue culture laboratories are freed of their constraints, particularly

financial and marketing. They are the businesses that would enable India retain

its self-sufficiency on agricultural production front. Finance being the heart of

any business, finances will have to be controlled and regulated systemically in

tissue culture business as well. A thorough study of the financial viability and

commercial prospects of the tissue culture venture should be carried out by the

new entrant. It is the need of the time with the backdrop of an era of

globalization and the emergence of World Trade Organizafion.

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