Hairy root cell culture


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Hairy root cell culture

  1. 1. By: anjali p. parab<br />HAIRY ROOT CELL CULTURE<br />
  3. 3. 3/30/2011<br />3<br />
  4. 4. Hairy root cultures:<br />Definition: <br /><ul><li>It is the culture produced after the infection of explants or cultures by the gram negative soil bacterium Agrobacteriumrhizogenes.
  5. 5. This processes take advantage of the naturally occurring hairy root disease in Dicotyledons.</li></ul>3/30/2011<br />4<br />
  6. 6. PROPERTIES OF HAIRY ROOTS <br /><ul><li>Genotype and phenotype stability
  7. 7. Autotrophy in plant hormones
  8. 8. Fast growth
  9. 9. High levels of secondary metabolite production</li></ul>3/30/2011<br />5<br />
  10. 10. 3/30/2011<br />6<br />
  11. 11. Production of hairy roots in vivo:<br />Agrobacterium recognizes some signal molecules exuded by wounded plant cells and becomes attached to it.<br />The bacteria contain the Root inducing plasmid (Ri-plasmid) <br />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:<br />3/30/2011<br />7<br />
  12. 12. Production of hairy roots in vivo:<br />Proliferate by increasing the rate of cell division (cytokine expression) and cell elongation (auxin expression) to produce the hairy roots. <br />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.<br />3/30/2011<br />8<br />
  13. 13. THE PROCESS…………<br />Plantcell<br />Agrobacterium cell<br />Ri-plasmid<br />3/30/2011<br />9<br />
  14. 14. Ri-Plasmid<br />Ri-Plasmid<br />3/30/2011<br />10<br />
  15. 15. Induction of hairy root cultures in vitro:<br />Explants are wounded and then inoculated with Agrobacteriumrhizogenes. <br />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.<br />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. <br />The decontaminated hairy roots can be subcultured on phytohormone-free medium.<br />3/30/2011<br />11<br />
  16. 16. Bioreactors for hairy root cultures<br /><ul><li>Stirred tank bioreactor
  17. 17. Air lift bioreactor
  18. 18. Bubble column bioreactor
  19. 19. Turbine blade bioreactor
  20. 20. Mist (Trickle bed) bioreactor
  21. 21. Rotating drum bioreactor
  22. 22. Spin filter bioreactor</li></ul>3/30/2011<br />12<br />
  23. 23. Stirred tank reactor with suspension culture of <br />Helianthus annuus<br />Bubble column with<br /> Hairy roots of B. vulgaris<br />3/30/2011<br />13<br />
  24. 24. Bubble column with Hairy roots of Harpagophytumprocumbens<br />3/30/2011<br />14<br />
  25. 25. Agar plate with Hairy roots of Beta vulgaris (beetroot) <br />3/30/2011<br />15<br />
  26. 26. Advantages of hairy root cultures:<br />The hairy root system is genetically and biosynthetically stable<br />High production of secondary metabolites. <br />The culture can grow under phyto-hormone-free conditions. <br />The culture shows fast growth which reduces the culture time and easy the handling<br />3/30/2011<br />16<br />
  27. 27. Application of hairy root cultures:<br />Functional analysis of genes.<br />Expressing foreign proteins.<br />Production of secondary metabolites.<br />The culture may produce compounds which is not found in untransformed roots.<br />The culture may change the composition of metabolites.<br />The culture can be used to regenerate a whole plant.<br />3/30/2011<br />17<br />
  28. 28. secondary metabolites produced in root culture<br />3/30/2011<br />18<br /><ul><li>L-DOPA: a precursor of catecholamines, an important neurotransmitter used in the treatment of Parkinson’s disease
  29. 29. Shikonin: used as an anti-bacterial and anti-ulcer agent
  30. 30. Anthraquinone: used for dyes and medicinal purpose
  31. 31. Opiate alkaloids: particularly codeine and morphine for medical purposes
  32. 32. Berberine: an alkaloid with medicinal uses for cholera and bacterial dysenterry
  33. 33. Valepotriates: used as a sedative
  34. 34. Ginsenosides: for medicinal purposes
  35. 35. Rosmarinic acid: for antiviral, suppression of endotoxin shock and other medicinal purposes
  36. 36. Quinine: for malaria
  37. 37. Cardenolides or Cardioactive glycosides: for treatment of heart disease</li></li></ul><li>3/30/2011<br />19<br /> 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)<br />
  38. 38. 3/30/2011<br />20<br />
  39. 39. REFERENCES<br /><ul><li>1)Pharmacognosy by Trease & Evans
  40. 40. 2)Plant tissue culture by Bhojwani & Razdan
  41. 41. 3)
  42. 42. 4)</li></ul>3/30/2011<br />21<br />
  43. 43. THANK YOU!<br />3/30/2011<br />22<br />