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ABT 401 Agrl Biotech/NM Boopathi/Lec 3 History of Tissue Culture

History of Plant Tissue Culture

Early concepts:

1838: Schwann and Schleiden

postulated Totipotency theory:

Cells have the capacity to

regenerate into complete plant.

1902: German botanist G.

Haberlandt developed the

concept of in vitro cell culture. He

was the first to culture isolated,

fully differentiated palisade cells of

Lamium purpureum in Knop’s

medium enriched with sucrose.

Cells enlarged but never divided.

Haberlandt is regarded as the

father of tissue culture.

1904: Hanning excised nearly

mature embryos of crucifers and

successfully grew them to maturity

on mineral salts and sugar

solutions. This is the first attempt

in embryo culture.

Root tip and embryo culture:

1922: Kotte and Robbins

simultaneously conceived that in

vitro culture could be made easier

by using meristematic cells such as

root tip or bud.

1925&1929: Laibach

demonstrated the practical

applications of zygotic embryo

culture in the field of plant

breeding.

1934: White developed a synthetic

medium which was later proved to

be one of the basic media for

variety of cell and tissue culture.

1969: Murashige and Skoog

proposed a media for tobacco

tissues. Still now this is the media

of preference with slight

modifications according to the

need.

1958: Reinert first reported

somatic embryo formation from

carrot tissue.

1959: Braun regenerated the first

plant from a mature plant cell.

Gautheret published first

extensive hand book in Plant

Tissue Culture.

Role of growth regulators:

1939: The possibility of culturing

plant tissues for an unlimited

period using media enriched with

growth regulator such as auxins

(Eg. IAA) was announced

independently by Gautheret,

White and Nobecourt. The

methods and media now used are

modifications of those established

by these pioneers.

1941: Van Overbeek

demonstrated for the first time the

stimulatory effect of coconut milk

(embryo sac fluid) on embryo


development and callus formation

in Datura.

1955: Miller separated the first

known cytokinin from the DNA

herring sperm and named it

kinetin.

1957: Skoog and Miller proposed

the concept of hormonal control of

organ formation. High

concentration of auxin promoted

rooting whereas proportionately

more kinetin initiated bud or shoot

formation.

Plant regeneration and

micropropagation:

1944: Skoog shown for a first time

that in vitro cultures of tobacco can

be used to study adventitious shoot

formation.

1946: Ball successfully raised

transplantable whole plants and

shown micropropagation.

Murashige has developed

standard methods of propagation

in large number of species and he

is intimately associated with this

technique.

1953: Muir demonstrated single

cell culture in Nicotiana tobaccum.

1965: Vasil and Hildebrandt

raised whole plants starting from

single cells of tobacco.

1980: Vasil and his group

demonstrated that embryogenic

cultures of most recalcitrant

cereals can be established using

immature embryos as explants.

Secondary metabolites:

1950-1960: G. N. Nickell, made

distinguished studies in secondary

metabolite production through

tissue culture. The first attempt for

the industrial production of

secondary metabolites in vitro was

made in this period by Pfizer

Company.

1983: The first tissue culture

commercial product, Shikonin, was

produced by Mitsui

Petrochemical Co, Japan, from

cell culture of Lithospermum

erythrorhizon.

Pathogen free plants:

1952: Morel and Martin

recovered virus free Dahlia and

potato plants from culture obtained

by cultivating the shoot meristem

of infected plants. The technique

allowed the production of an

estimated 4 million genetically

identical plants from a single bud in

a period of 1 year.

1975: Hendre and Co-workers

established technique for obtaining

virus free Sugarcane, Citrus, Potato

and Cassava.


Genetic Variability:

1955: Gautheret and Nobecourt

shown independently that tissue

cultures are a direct source of

genetic variability.

1977: Heinz and Mee reported

variation in sugarcane hybrids

regenerated from cell cultures, for

a first time.

1981: Larkin and Scowcroft,

proposed the term somoclonal

variation.

In vitro culture of reproductive

organs:

1960: Kanta (India) demonstrated

successful test tube fertilization in

Papaver rhoeas for a first time (in

vitro pollination).

1966: Guha and Maheswari

(India) first produced haploid

Datura plant from pollen grains.

Normally, somatic cells of higher

plants have a diploid chromosome

number while reproductive cells

(gametes) are haploid.

1976: San Noeum reported her

first result on induction of haploid

plants (parthenogenetic or

apogamous) from unpollinated

ovaries through in vitro culture of

ovary isolated from Hordeum

vulgare.

1990: Kranz et al., reported

electrofusion of isolated male and

female gametes of maize and plant

regeneration from the fusion

product. This is the first

demonstration of in vitro

fertilization in higher plants.

Protoplast culture:

1960: Cocking produced large

quantities of protoplasts by using

cell wall degrading enzymes

(Cellulase).

1972: Carlson obtained first

somatic hybrid by fusion of

protoplasts isolated from Nicotiana

galuca and N. langsdorffii. 1979:

Melchers shown Somatic

hybridization of potato and tomato

by protoplast fusion.

1982: Zimmermann fused

protoplasts by electrical stimulus.

Germplasm storage:

1973: Nag and Street, first

reported the possibility of

regeneration of plants from carrot

cells frozen at the temperature -

196 °C of liquid nitrogen.

1985: Kitto and Janick produced

artificial seeds.

Genetic Transformation:

1907: Smith and Townsend has

shown that Agrobacterium

tumefaciens causes crown gall

disease in some plants.

1977: The utility of the bacteria as

a gene transfer system in plants


was first recognized by Chilton

and his group.

1983: Barton demonstrated that

heterologous DNA inserted into the

T-DNA of Ti plasmid could be

transferred to the plant.

1984: Horsch et al., produced the

first transgenic tobacco plants

expressing engineered foreign

genes were produced using A.

tumefaciens.

1986: Abel and his group produced

the first transgenic plants using

microprojectile bombardement or

biolistic method (Particle gun).

Cellular Totipotency:

The inherent potentiality of a

plant cell to give rise to a whole

plant is described as cellular

totipotency. This is a capacity

which is retained even after a cell

has undergone final differentiation

in the plant body. In plants, even

highly mature and differentiated

cells retain the ability to

regenerate to a meristematic state

as long as they have an intact

membrane system and a viable

nucleus. This is contradicting to

animals, where differentiation is

generally irreversible.

For a differentiated cell, to

express its totipotency, it first

undergoes dedifferentiation

followed by redifferentiation. The

phenomenon of a mature cell

reverting to the meristematic state

and forming undifferentiated callus

tissue is termed ‘dedifferentiation’.

The phenomenon of conversion of

component cells of callus tissue to

whole plant or plant organs is

called as ‘redifferentiation’.

Explant and explant selection:

A plant organ or piece of

tissue used to initiate a culture.

Almost all parts of plant are

amenable to in vitro plant

regeneration (E.g. Tobacco).

However, in certain plants some

organs may be more regenerative

than the others (e. g. in Glycine

max, the hypocotyls exhibits higher

potentiality for shoot formation

than the root segments).

The regenerability of an explant is

influenced by several factors:

1. Organ from which it is

derived.

2. The physiological state of

explant

3. Size of the explant

4. Orientation of the explant on

the medium and


5. Its inoculation density.

Sterilization and its

techniques:

Sterilization is the process of

inactivating or removing all living

organisms from a substance or

surface. Different kinds of

sterilization procedures were

adapted in plant tissue culture.

They are,

1. Heating:

Dry heat (e.g. Hot air

oven) is used for glassware and

Steam or moist or wet heat

(e.g. Autoclave) is used for

sterilization of media and

fermenter vessels.

2. Radiation:

UV radiation and high efficiency

particulate air (HEPA) filter in

laminar air-flow chamber used for

sterilization of transfer area.

3. Chemicals:

95 % ethanol used for sterilizing

forceps, scalpels, needles etc.,

Hypochlorite solutions (sodium or

calcium) and antibiotics are used

as sterilizing agents to disinfect

plant tissues.

4. Ultra filtration:

The solution of thermolabile

compounds (e.g. vitamins,

antibiotics etc) is sterilized by

membrane filtration.

Do you know….

The difference between somatic

embryo and zygotic embryo?

What temperature and pressure is

used in normal autoclaving

procedure?

Further readings…

Bhojwani. S.S. and Razdan, M.K.

2004. Plant Tissue Culture:

Theory and Practice, a revised

edition. Elsevier, Amsterdam.

Razdan, M. K. 1994. An

introduction to plant tissue

culture. Oxford and IBH publishing

company, New Delhi.

Singh, B. D. 2003. Biotechnology.

Kalyani publishers, New Delhi.

Slater, A., N. W. Scott and Mark, R.

Fowler. 2003. Plant

Biotechnology: the genetic

manipulation of plants. Oxford

University press Inc., New York.

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