Hairy Root Cell Culture

Presented by Abu RaihanM.Tech, 1st year,Biotechnology

Culture Types

Callus Culture

Cell Suspension Culture

Protoplast Culture

Hairy Root Culture

Hairy Root Culture

Definition:

It is the culture produced after the infection of explants andcultures by the gram negetive Soil bacterium Agrobacterium rhizogenes.

This processes take advantage of the naturally occuring hairyroot disease in Dicotyledons.

Properties of hairy roots

Genotype and phenotype stability

Autotrophy in plant hormones

Fast growth

High levels of secondary metabolite production

Production of hairy root in plant cell

Agrobacterium recognizes some signal molecules exuded by wonded plant cells andbecomes 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.

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.

The Process

Ri-plasmid

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.

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

Bubble column with hairy root culture picture

Agar plate with hair root picture

Hairy root cell culture picture

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.

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.

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