This document discusses hairy root cell culture. Hairy root culture involves infecting plant explants with the soil bacterium Agrobacterium rhizogenes, which transfers genes to the plant genome and causes roots to form with increased cell division and elongation, producing "hairy roots". Hairy roots have properties like genotype/phenotype stability, fast growth, and high production of secondary metabolites. The process involves wounding explants, inoculating with A. rhizogenes, inducing hairy roots within 1 week to 1 month, and subculturing in antibiotic media to remove bacteria. Hairy roots can be cultured in various bioreactors and have applications like gene analysis, protein expression, and secondary metabolite production.

1. Hairy Root Cell Culture
Presented by Abu Raihan
M.Tech, 1st year,Biotechnology

2. Culture Types
Callus Culture
Cell Suspension Culture
Protoplast Culture
Hairy Root Culture

3. Hairy Root Culture
Definition:
It is the culture produced after the infection of explants and
cultures by the gram negetive Soil bacterium
Agrobacterium rhizogenes.
This processes take advantage of the naturally occuring hairy
root disease in Dicotyledons.

4. Properties of hairy roots
Genotype and phenotype stability
Autotrophy in plant hormones
Fast growth
High levels of secondary metabolite
production

5. Production of hairy root in plant cell
Agrobacterium recognizes some signal
molecules exuded by wonded plant cells and
becomes attached to it.
The bacteria contain the Root inducing
plasmid ( Ri-plasmid).
The bacteria Genetically trasfer part of
the Ri-plasmid called the trasfer DNA
(T-DNA) to the plant genome.

6. Production of hairy root in plant cell
The plant cell to proliferate by increasing
the rate of cell division ( cytokine expression )
and cell elongation (auxin expression) to
produce the hairy roots.
Produce the opines which is a type of unusual
amino acids ( octopine, agropine, nopaline,
mannopine amd cucumopine) which is used
by the bacterium as a carbon , nitrogen and
energy source.

7. The Process

8. Ri-plasmid

9. Induction of hairy root cultures In Vitro
Explants are wounded and then inoculated with
Agrobacterium rhizogenes.
Usually two or three days later, the explant can
be transferred into solid media with antibiotics,
such as cefotaxime , vancomycine or ampicillin to
kill or eliminate redundant bacteria.
The hairy roots will be induced within a short
period of time, which varies from one week to
over a month depending on different plant
species.
The decontaminated hairy roots can be
subcultured on phytohormone –free medium.

10. Bioreactors for hairy root cultures
Stirred tank bioreactor
Air lift bioreactor
Bubble column bioreactor
Turbine blade bioreactor
Trickle bed bioreactor
Rotating drum bioreactor
Spin filter bioreactor

11. Bubble column with hairy root culture picture

12. Agar plate with hair root picture

13. Hairy root cell culture picture

14. Advantages of hairy root cultures
The hairy root system is genetically
and biosynthetically stable.
High production of secondary
metabolites
The culture can grow under phyto-hormone-
free conditions.
The culture shows fast growth which
reduces the culture time and easy
the handling.

15. Application of Hairy Root Cultures
Functional analysis of genes.
Expression foreign proteins.
Production of secondary metabolites.
The culture may produce compounds
which is not found untransformed roots.
The culture may change the composition
of metabolites.
The culture can be used to regenerate a
whole plant.

16. Thanks