Plant Cell and Tissue Culture Technology:

General Introduction
 What is plant tissue culture?

Tissue culture
Ti lt is
i the
th term
t usedd for
f “the
“th process off growing
i cells
ll
artificially in the laboratory”

Tissue culture involves both pplant and animal cells

Tissue culture produces clones, in which all product cells have the
same genotype (unless affected by mutation during culture).
 Tissue culture had its origins
g at the
beginning of the 20 th century with
the work of Gottleib Haberlandt
(plants)

and Alexis Carrel (animals)

Gottleib Haberlandt
 What’s the History?
1902 – Haberlandt conducted the first but unsuccessful attempt of tissue culture using 
monocotyledons. His idea was to achieve continued cell division in explanted tissue grown 
on nutrient medium 

1926 ‐ FW Went demonstrated that there were growth substances in coleoptiles 
from oats

A. Germination of an oat seed 
B. Decapitate tip of coleoptile
p p p and 
place on agar block.
C. Agar block is placed on top of the 
decapitated tip of the seedling (what 
happens?).
happens?)
D. Agar block is placed on the side of 
the leaf on the decapitated tip of the 
seedling (what happens?).
 What’s the History?
1934 ‐ White generated continuously growing culture of meristematic cells of 
tomato on medium containing salts, yeast extract, sucrose and 3 vitamins 
(pyridoxine, thiamine, nicotinic acid). This established the importance of 
additives
dditi i ti
in tissue culture
lt

The development of improved nutrient solutions, use of the newly discovered 
plant growth regulator indole‐3‐acetic acid (IAA), informed choice of plant 
material, and appreciation of the importance of aseptic cultures, led to 
sustained growth of root tips, and carrot root and tobacco stem tissues, for 
indefinite periods of time (Gautheret 1934, 1939, 1985; Nobe´court 1939; 
White 1939)
White 1939).

1953 ‐ Miller and Skoog discovered kinetin, a cytokinin that plays an active 

role in organogenesis
1954 – First plant grown from a single cell

1957 ‐ Discovery that root or shoot formation in culture depends on 
auxin: cytokinin ratio
auxin: cytokinin
1962 ‐ Development of the Murashige and Skoog (MS) medium
 What is plant tissue culture?

The growth or regeneration of plant cells, tissues, organs or 
h h f l ll
whole plants in artificial medium under aseptic conditions
 Totipotency?

There are numerous methods to propagate plants in tissue culture.

But the one principle that is constant is totipotency
But the one principle that is constant is totipotency – all plants and plant 
all plants and plant
parts have this potential.

Totipotency is the genetic potential of a plant cell to produce the 
entire plant. In other words, totipotency is the cell characteristic in 
which the potential for forming all the cell types in the adult 
organism is retained.

Expression of totipotency
Expression of totipotency depends on competence, by which the ability of cells to 
depends on competence, by which the ability of cells to
be induced along a particular developmental pathway and determination, in which 
cells become irreversibly committed to a particular pathway. To express totipotency
by a mature differentiated cell, it first undergoes de‐differentiation followed by re‐
diff
differentiation
ti ti
 Basic Plant Development & Tissue culture?

reversible
Differentiation
De‐differentiation

Irreversible
Re‐differentiation
SEED Callus: a mass of 
highly dividing 
Plant undifferentiated cells
Totipotency:
The ability of a cell to differentiate and develop into a whole plant when 
g
given the correct conditions. This is because every cell has the genetic 
y g
potential of the parent plant.

Gene transfer
Plant Cell

Plant tissue T
Transgenic plant
i l
Callus

Plant Organ Somatic embryo
Cell suspension culture

Production of value 
Production of value
added secondary 
New Plant metabolites
 Terminology

• Explant
– Living tissue transferred 
from a plant to an artificial 
medium for culture
medium for culture. 

– It can be any portion of 
the shoot leaves roots
the shoot, leaves, roots, 
flower or cells from a plant.
 Differentiation:
The cells derived from root apical meristem (RAM) and shoot apical meristem
The cells derived from root apical meristem (RAM) and shoot apical meristem
(SAM) and cambium, differentiate, mature to perform specific functions. This act 
leading to maturation is termed differentiation. They, undergo a few or major 
structural changes both in their cell walls and protoplasm.

Example: Tracheids are elongated cells in the xylem of vascular plants that serve in the
transport of water and mineral salts. Tracheids are one of two types of tracheary
elements, vessel elements being the other. Tracheids, unlike vessel elements, do not
have perforation plates.
All tracheary elements develop a thick lignified cell wall, and at maturity
the protoplast has broken down and disappeared. The presence of tracheary elements
i the
is th defining
d fi i characteristic
h t i ti off vascular
l plants
l t to
t differentiate
diff ti t them
th f
from non‐vascular
l
plants.

Irreversible

Meristem cell No protoplast

De‐differentiation: The living differentiated cells, that by now have lost the capacity 
to divide can regain the capacity of division under certain conditions This
to divide can regain the capacity of division under certain conditions. This 
phenomenon is termed as de‐differentiation. For example, formation of meristems –
interfascicular cambium and cork cambium from fully differentiated parenchyma 
cells. While doing so, such meristems / tissues are able to divide and produce cells 
that once again lose the capacity to divide but mature to perform specific functions, 
i.e., get redifferentiated.

In tissue culture you can change 
y g
permanent cell into meristmatic 
cell by applying artificial conditions

Growth 
regulator 
and other 
conditions

Callus
Plant callus is a growing and actively dividing mass of unorganized 
plant  parenchyma cells. 

In living plants, callus cells are those cells that cover a plant wound

Wound callus Tissue culture callus

X
Re differentiation: While doing so, such meristems
Re‐differentiation: While doing so such meristems / tissues are able to 
/ tissues are able to
divide and produce cells that once again lose the capacity to divide but 
mature to perform specific functions, i.e., get re‐differentiated and this 
process is called re‐differention.

re‐differention.

de‐differention.

Growth regulator & 
other conditions
 Natural differentiation (ND)  vs re‐differentiation (RD)

Shoot and root will 
Sh t d t ill
come together

ND

Shoot Root
Regeneration

Root Shoot

RD
Is plant cells are really 
Totipotent?
 Aseptic: Free from contamination by microorganisms such as bacterial, 
fungi, and viruses – For surroundings

Axenic: Free from contamination by microorganisms such as bacterial, 
fungi and viruses – for culture / biological part
fungi, and viruses  for culture / biological part