hairy root culture

1. PROTEGES-Kartikey Vikram Singh
And
Vineet Dubey
MENTOR-Santosh Kr. Mishra

2. 

Recently there has been a perceptible revival of
interest in the use of medicinal plants
throughout the world culminating into a
manifold increase of medicinal plant based
products.
At present there are about 130 clinically useful
prescription drugs of known chemical
structures, solely derived from higher plants
and used in modern system of medicines.

3. 
In view of the spectacular increase in the
demand of plant based compounds, the
concerned medicinal plants have been
indiscriminately exploited leading to scarcity
or extinction of many valuable plant species.
plant species
product
Cost($/kg)
Cath. roseus
Ajmalicine
37,000
P. somniferum
Codeine
17,000
Cath. roseus
Vincristine
20,00,000
Cath. roseus
Vinblastine
10,00,000
Colchium autumnale
colchicine
35,000
Panax ginseng
Ginsenosides
50,000

4. 


This project deals with the production of highvalue secondary metabolites including
pharmaceuticals and food additives through
root cultures and transgenic roots obtained
through biotechnological means.
Recent developments in transgenic research
have opened up the possibility of the metabolic
engineering of biosynthetic pathways to
produce high-value secondary metabolites.

5. 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.

6. 

Agrobacterium
recognizes
some
signal
molecules exuded by wounded plant cells and
becomes attached to it.
The bacteria contain the Root inducing plasmid
(Ri-plasmid) The bacteria genetically transfer
part of the Ri-plasmid called the transfer DNA
(T-DNA) to the plant genome.

7. 

where it gets expressed and make 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, and cucumopine) which is used by
the bacterium as a carbon, nitrogen and energy
source.

10. 



The hairy root system is genetically and
biosynthetically stable.
High production of secondary metabolites.
The culture can grow under phyto-hormonefree conditions.
The culture shows fast growth which reduces
the culture time and easy the handling

11. 1.
2.
3.
4.
5.
The main aim of this project is to synthesize secondary
metabolites from hairy root culture of medicinal plants
this process will comprise of following stages:
Selecting a plant species appropriate for the desired
product.
Through Micropropagation obtaining an infection free
sample.
By means of genetic transformation induce the hairy
root culture.
Obtaining the product and analysing the final product
for quality.
Observing effect of different chemicals on the culture.

12. Time
Objective
two month
Literature and Review
two month
Micro propagation of the Selected
Plant
two month
Biotransformation
two month
Optimisation of different parameters
of culture
two month
effect of different types of chemical
factor on hairy root culture
two month
Analysis of secondary metabolites
produced by hairy roots

13. 



To grow the culture of A. rhizogenes we can
prepare media either LB or YEB.
To grow the hairy root we are preparing MS
media and enzyme supplement stock solutions.
Establishment of culture room is in progress.
Selection of the plant species for micro
propagation.

14. References:



S. Ramachandra Rao and G.A. Ravishankar;
Biotechnology advances 20 (2002) 101 – 153; Plant cell
cultures: Chemical factories of secondary metabolites.
Suchitra Banerjee, Sailendra Singh, Laiq Ur Rahman;
Biotechnology advances 30 (2012) 461–468;
Biotransformation studies using hairy root cultures.
Flores HE, Dai YR, Freyer AJ, Michaels PJ.
Biotransformation of menthol and geraniol by hairy
root cultures. Plant Physiol Biochem 1994;32:511–9.
Jung G, Tepfer D. Use of genetic transformation by the
T-DNA of Agrobacterium rhizogenesto stimulate
biomass and tropane alkaloid production in Atropa
belladonna and Calystegiasepium roots grown in vitro.
Plant Sci1987;50:145 – 51.

15. 



Ambros PF, Matzke AJM, Matzyke MA.
Localization of Agrobacterium rhizogenes T-DNA
in plant chromosomes by in situ hybridization.
EMBO J 1986;5:2073 – 7
Newman DJ, Cragg GM. Natural products as
sources of new drugs over the last 25 years.J Nat
Prod 2007;70:461–77
Subroto MA, Kwok K, Hamill JD, Doran PM.
Coculture of genetically transformed roots and
shoots for synthesis, translocation and
biotransformation of secondary metabolites.
Biotechnol Bioeng 1996;49:481 – 94.
Banerjee S, Shang TQ, Wilson AM, Moore AL,
Strand SE, Gordon MP, et al. Expression of
functional mammalian P450 2E1 in hairy root
cultures. Biotechnol Bioeng 2002;77:462–6.