Hairy root cell culture

an attempt to provide brief data on the topic!

1. By: anjali p. parab HAIRY ROOT CELL CULTURE

2. HAIRY ROOT CELL CULTURE

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4. 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 . 3/30/2011 4

5. PROPERTIES OF HAIRY ROOTS Genotype and phenotype stability Autotrophy in plant hormones Fast growth High levels of secondary metabolite production 3/30/2011 5

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7. Production of hairy roots in vivo : 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, where it gets expressed and make the plant cell to: 3/30/2011 7

8. Production of hairy roots in vivo : 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. 3/30/2011 8

9. THE PROCESS………… Agrobacterium cell Plant cell Ri-plasmid 3/30/2011 9

10. Ri -Plasmid Ri-Plasmid 3/30/2011 10

11. 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 , vancomycin 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. 3/30/2011 11

12. 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 3/30/2011 12

13. Bubble column with Hairy roots of B. vulgaris Stirred tank reactor with suspension culture of Helianthus annuus 3/30/2011 13

14. Bubble column with Hairy roots of Harpagophytum procumbens 3/30/2011 14

15. Agar plate with Hairy roots of Beta vulgaris (beetroot) 3/30/2011 15

16. 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 3/30/2011 16

17. Application of hairy root cultures: Functional analysis of genes. Expressing foreign proteins. Production of secondary metabolites. The culture may produce compounds which is not found in untransformed roots. The culture may change the composition of metabolites. The culture can be used to regenerate a whole plant. 3/30/2011 17

18. secondary metabolites produced in root culture 3/30/2011 18 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

19. 3/30/2011 19 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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21. REFERENCES 1) Pharmacognosy by Trease & Evans 2)Plant tissue culture by Bhojwani & Razdan 3)www.springerlink.com 4)www.420magazine.com 3/30/2011 21

22. THANK YOU! 3/30/2011 22