1. Plant tissue culture is a set of techniques used to propagate plants under sterile conditions on artificial nutrient media. 2. Meristematic cells have the ability to differentiate into all plant cell types and can be used to regenerate an entire plant through tissue culture techniques. 3. The basic steps of micropropagation include sterilization, establishment of explants on nutrient media, shoot multiplication, and rooting to regenerate whole plants that can be hardened off and transferred to soil conditions.

1. Presented
by
Dr. C. SOUNDAR RAJU
Assistant Professor
VIVEKANANDA COLLEGE
(Residential & Autonomous)

2. We know about plant regeneration
 Plant propagation is the process of creating new plants
 Plant propagation can also refer to the artificial or natural
dispersal of plants.

3. Vegetative propagation (asexual reproduction)

4. MONOCOT
DICOT
Seed propagation (sexual reproduction)

5. Meristem
 Meristematic cells (undifferentiated cells) capable of cellular differentiation.
 Meristematic tissue is made up of small cells that have thin walls and large nuclei.
 The cells have no vacuoles and intercellular spaces.
 It can be induced to reproduce an entire plant.
 Shoot apical meristem (SAM)- shoot, leaves and flowers.
 Root apical meristem (RAM)- root growth.
 Further developed into permanent tissues
 Protective tissue, parenchyma tissue,
 Sclerenchya tissue, collenchyma tissue,
 Xylem tissue and phloem tissue.
 Meristem- merizein (greek word ) meaning to divide, in recognition of its inherent
function (1858)
Carl Wilhelm von Nägeli
(1817 – 1891)
How is grow ?

6. What is differentiation ?

8.  It is the techniques
 Plants raised under controlled condition
 Plants are regenerated from cell, protoplast, tissue, organ (leaf, stem, root, flower
part) etc.,
 They were cultured on artificial nutrient medium
 Incubated at aseptic condition (pathogenic free / controlled temperature,
photoperiod etc)
Plant tissue culture Explant
Gottlieb Haberlandt (1854-1945)
(Father of Tissue Culture)

9. Micropropagation
Temperature : 25 ± 2 ºC
Photoperiod : 16 h / 8 h
(light / dark)

10. Inherent capacity of plant
cell to develop into
complete plant
Totipotency
Dedifferentiation:
Mature cell reverting back to
meristamatic state and forming an
undifferentiated mass of cells (callus)
Callus:
Undifferentiated cells, non functional
cells.
Redifferentiation:
The ability of dedifferentiated cell/callus
to form a whole plant.

11. Artificial medium
 Macronutrient
Nitrogen, phosphorus, potassium, magnesium, calcium & sulphur
 Micronutrient
Manganese, iodine, copper, cobalt, boron, molybdenum, iron & zinc
 Organic supplement (vitamin)
Mitol, Nicotinic acid, Pyridoxine-HCl, Thiamine-HCl, Glycine
 Carbon source
Sucrose
 Plant growth regulators
Auxin- IAA, IBA, NAA, 2,4-D
Cytokinin- Kinetin, Zeatin, BA, Thidiazuron
Gibberellins- GA3
 Gelling agent (except liquid medium)
Agar
 pH
5.6-5.8

12. Elements important for plant nutrition and their physiological function

13. MS medium
Murashige, T. and Skoog, F.
Composition of commonly used plant tissue media

14. Auxin
IAA
IBA
NAA
Cytokinin
Kinetin
Zeatin
BA
TDZ
.
.
.
.
.
.
.

16. Sterilization
Autoclave
121 ºC, 15 min

17. Surface sterilization

18. UV- 15-20min
Tissue culture room (25 ± 2 ºC)
Laminar air flow chamber
In vitro
establishment
Field transformationHardening

19. Organogenesis
Direct
organogenesis
Indirect
organogenesis
Somatic
embryogenesis
Direct somatic
embryogenesis
Indirect somatic
embryogenesis
Plant regeneration
The rearing of plant cell / tissue / organ / callus / protoplast into young plants

20. Direct method
Indirect method

21. Shoot multiplication

22. Analysis of genetic stabilityMolecular markers
 RAPD, RFLP, SSR, ISSR
PCR

23. Direct organogenesis

24. Indirect organogenesis

25. Induction of callus
MS salts + 2,4-D

27. somatic embryogenesis

28. Direct somatic embryogenesis

30. Indirect somatic embryogenesis

31. Synthetic seed production

32. Protoplast culture & somatic hybridization
 Present cytoplasm and nucleus
 Cell wall absent
Washing sloution
Dihydrogen Potassium orthophophate
Potassium nitrate
Calcium chloride dihydride
Megnesium sulphate hepta hydrate
Potassium iodide
Copper sulphate pentahydrate
Sucrose
pH 5.7

33. Liquid culture
Medium does not contain solidifying agent

34. Cell suspension culture

35. Callus

36. Gene transformation
 The first genetically modified crop plant was produced in 1982, an antibiotic-resistant
tobacco plant
 The first field trials occurred in France and the USA in 1986, when tobacco plants
were engineered for herbicide resistance

37. Southern blot analysis

40. Rhizogenesis
Induction of roots
Hairy root culture
Agrobacterium rhizogenes

41. Industrial application

49. Applications

50. Thanking you