1 A III

Totipotency is a concept!
TOTIPOTENCY & TISSUE CULTURE
(of a cell or part) having the
potential for developing in various
specialized ways in response to
external or internal stimuli.

Totipotency is the ability of a

single cell to divide and produce all
of the differentiated cells in an
organism.
 Spores and zygotes are examples
of totipotent cells.
 In the spectrum of cell
potency, Totipotency represents
the cell with the greatest
differentiation potential.
TOTIPOTENCY

The ability of cells such as an

embryonic stem cell to differentiate
into any type of body cell.
 TISSUE CULTURE
Tissue culture is a process that involves exposing
 Tissue culture is a plant tissue to a specific regimen of nutrients, hormones,
and light under sterile, in vitro conditions to produce many
technique new specific tissue, each a clone of the original mother
tissue, over a very short period of time.
 Used to maintain or grow
specific cells, tissues or
organs under sterile
conditions on a
nutrient culture medium
of known composition.
OTHER EQUIPMENT

• Seeds of white • weighing scales,

• mustard, • plastic tray,
• agar, • 250ml beaker,
• distilled water, • glass rod,
• damp sponge,
• scissors,
• cling film,
• sunny window sill
• McCartney bottles,
METHOD:

 sprinkle seeds on damp sponge and allow to germinate.

 Use when just starting to unfold their cotyledons (seed leaves).
 Make up Agar gel and pour 2cm height of gel into McCartney bottles and allow to set.
 With sharp scissors, cut the tops off just below the shoot apex (including the cotyledons).
 This is called an explants.
 Push the stem of the explant into the gel (making sure cotyledons don’t touch agar) cover with cling film and
place on sunny windowsill.
 Observe over 10 days.
OUTCOME:

 explant grows roots and leaves continue to grow.

Question (Homework):
Explain why they are covered in cling film and why they continue to grow
even when covered. Also why they shouldn’t be opened again.
POSSIBLE EVALUATION ISSUES

 Unwanted pathogens growing in the gel as it is a good source of

water and nutrients
 Wrong part of the plant cut and inserted into gel.
INVESTIGATE THE CONDITIONS NEEDED FOR THE
GROWTH OF PLANT CELLS.
USING TISSUE CULTURE TECHNIQUES
** NOTE: PLANT CELLS CAN GROW TO FORM A CALLUS, A MASS OF UNDIFFERENTIATED CELLS
CALLUS
In plants parench
yma is the ground
tissue of non-
 Plant woody structures.
callus (plural calluses or calli)
 Is a growing mass of
unorganized
plant parenchyma cells.
 In living plants, callus cells are
those cells that cover a plant
wound.
THE MAIN INGREDIENTS NEEDED FOR THE GROWTH OF CALLUSES
ARE:

 • mineral salts
 • auxin (IAA)
 • cytokinin
 • sugar, such as sucrose or glucose
OBJECTIVE:

 Using tissue culture techniques …

 Investigate the conditions needed for the growth of plant cells.
 Plant cells can grow to form a callus, a mass of undifferentiated cells.
 Investigate the effect of glucose concentration on the growth of calluses
from pieces of cotton plant tissue
PROCEDURE

 He mixed the ingredients listed above with an agar solution to make a gel on which he could grow the calluses.
 Working in sterile conditions, he removed small pieces of tissue, known as explants, from one cotton plant.
 The explants were placed on this gel.

Is a cell, organ, or piece of tissue which has been

transferred from plants and animals into a nutrient
medium
REPEAT

 This procedure was repeated using agar gel containing different

concentrations of glucose.
DATA COLLECTION

 After two weeks,

Data – 1:recorded the percentage of explants that had formed
calluses
Data – 2: the dry mass of the callus.
DEPENDENT VARIABLES ?
DEPENDENT VARIABLES

 percentage of explants {showing growth / forming calluses}

 dry mass of calluses
INDEPENDENT VARIABLES ?
INDEPENDENT VARIABLES

 glucose (concentration / percentage)