BY: ANJALI P. PARAB HAIRY ROOT CELL CULTURE
Hairy root cell culture
May 10, 2015
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BY: ANJALI P. PARAB
HAIRY ROOT CELL CULTURE
HAIRY ROOT CELL CULTURE
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Culture Types
CallusCell
suspension culture
Protoplast Hairy root
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HAIRY ROOT CULTURES:
Definition:
• It is the culture produced after the infection of explants or
cultures by the gram negative soil bacterium Agrobacterium
rhizogenes.
• This processes take advantage of the naturally occurring hairy
root disease in Dicotyledons.
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PROPERTIES OF HAIRY ROOTS
Genotype and phenotype stability Autotrophy in plant hormones Fast growth High levels of secondary metabolite
production
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PRODUCTION OF HAIRY ROOTS IN VIVO:
a) Agrobacterium recognizes some signal molecules exuded by
wounded plant cells and becomes attached to it.
b) The bacteria contain the Root inducing plasmid (Ri-plasmid)
c) The bacteria genetically transfer part of the Ri-plasmid called the
transfer DNA (T-DNA) to the plant genome, where it gets
expressed and make the plant cell to:
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PRODUCTION OF HAIRY ROOTS IN VIVO:
1. Proliferate by increasing the rate of cell division (cytokine
expression) and cell elongation (auxin expression) to produce the
hairy roots.
2. Produce the opines which is a type of unusual amino acids
(octopine, agropine,nopaline, mannopine, and cucumopine) which
is used by the bacterium as a carbon, nitrogen and energy source.
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THE PROCESS…………
Agrobacterium cell Plant cell
Ri-plasmid
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RI-PLASMID
Ri-Plasmid
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INDUCTION OF HAIRY ROOT CULTURES IN VITRO:
1. Explants are wounded and then inoculated with Agrobacterium rhizogenes.
2. Usually two or three days later, the explant can be transferred into solid
media with antibiotics, such as cefotaxime, vancomycin or ampicillin to kill
or eliminate redundant bacteria.
3. 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.
4. The decontaminated hairy roots can be subcultured on phytohormone-free
medium.
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BIOREACTORS FOR HAIRY ROOT CULTURES Stirred tank bioreactor Air lift bioreactor Bubble column bioreactor Turbine blade bioreactor Mist (Trickle bed) bioreactor Rotating drum bioreactor Spin filter bioreactor
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Bubble column with Hairy roots of B. vulgaris
Stirred tank reactor with suspension culture of Helianthus annuus
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Bubble column with Hairy roots of Harpagophytum procumbens
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15Agar plate with Hairy roots of Beta vulgaris (beetroot)
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ADVANTAGES OF HAIRY ROOT CULTURES:
1. The hairy root system is genetically and biosynthetically stable
2. High production of secondary metabolites.
3. The culture can grow under phyto-hormone-free conditions.
4. The culture shows fast growth which reduces the culture time
and easy the handling
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APPLICATION OF HAIRY ROOT CULTURES:
1. Functional analysis of genes.
2. Expressing foreign proteins.
3. Production of secondary metabolites.
4. The culture may produce compounds which is not found in untransformed
roots.
5. The culture may change the composition of metabolites.
6. The culture can be used to regenerate a whole plant.
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SECONDARY METABOLITES PRODUCED IN ROOT CULTURE
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L-DOPA: a precursor of catecholamines, an important neurotransmitter used in the treatment of Parkinson’s disease
Shikonin: used as an anti-bacterial and anti-ulcer agent Anthraquinone: used for dyes and medicinal purpose Opiate alkaloids: particularly codeine and morphine for medical purposes Berberine: an alkaloid with medicinal uses for cholera and bacterial dysenterry Valepotriates: used as a sedative Ginsenosides: for medicinal purposes Rosmarinic acid: for antiviral, suppression of endotoxin shock and other medicinal purposes Quinine: for malaria Cardenolides or Cardioactive glycosides: for treatment of heart disease
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30-L bioreactor cultures of medicinal plant - hairy roots of Atropa belladonna (upper left, start to culture; upper right, 3-week old; bottom, recovery of the 4-week old hairy roots)
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REFERENCES
1)Pharmacognosy by Trease & Evans 2)Plant tissue culture by Bhojwani &
Razdan 3)www.springerlink.com 4)www.420magazine.com
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THANK YOU!
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