History of plant tissue culture and journey through scientific inventions which laid idea on plant tissue culture and its nourishment

1. History of Plant tissue culture
KIRAN K.M.
2022-21-029
Ph.D. (Genetics and plant
breeding)
KERALA AGRICULTURAL UNIVERSITY
College of Agriculture, Padannakkad
Department of Genetics and Plant breeding
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2. Introduction
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Plant tissue culture, or the aseptic culture of cells, tissues, organs, and their
components under defined physical and chemical conditions in vitro, is an
important tool in both basic and applied studies as well as in commercial
application.

3. Birth of the concept
• The principles of tissue culture were contained in
the cellular theory, expressed in 1838-1839 by
Schleiden and Schwann.
• 1853- A. Trecul. performed experiment on callus formation by
decorticated trees such as Robinia, Pawlonia and Ulmus.
• 1759- Louise Duhamel : Invitro experiments on Wound healing in
plants
He removed bark of the elm tree  Observed callus formation in the
wounded region
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4. Conti..
• 1878- H. Vochting : Proposed polarity in development of
buds(Brassica rapa)
Upper portion of a stem piece : Shoot
Lower portion of a stem piece : Root
“Plants contain organ forming substances which are polarly
distributed” (Sachs 1880-1882)
• 1885- W. Z. Roux :The first experimental step in tissue culture
A fragment of the neural plate of a chick embryo removed and
cultivated in warm salt solution.
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(Gautheret, 1983)

5. • 1893- C. Rechinger : Determine the limits of divisibility of plant
materials
Conclusion : Pieces thicker than 1.5 mm would develop
The first approach of Plant tissue culture
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• 1902- G. Haberlandt – Father of plant tissue culture
Paper : "Experiments on the culture of isolated plant cells”
Work with single cells (palisade cells, pith cells, glandular hairs,
stamen hairs of Tradescantia:)
(Gautheret, 1983)

6. • Knop's nutrient solution, sucrose, asparagine and peptone
served as nutrients
Conspicuous cell growth, but no cell division
Used pieces of mature potato tuber
Indirect approach of tissue culture : cell division was controlled
by two hormones  Leptohormone & wound hormone
https://en.wikipedia.org/wiki/File:Gottlieb_Haberlandt.jpg
Gottlieb Haberlandt
Proposed concept of totipotency : “Theoretically all plant cells are able to give
rise to a complete plant”
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Conti..
• 1904- Hanning: Successfully cultured mature embryo of some crucifers
(Gautheret, 1983)

7. Developments in Animal tissue culture
• 1907- R. G Harrison : Cultivated the neuroblast of the frog in clotted
lymph
• 1910- A. Carrel established the present method of cultivating excised
animal tissues in a nutrient made of blood plasma and embryo juice or
their equivalents
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8. Root tip culture
• 1922- W. Kotte : succeeded in cultivating small root tips of pea and
maize in various nutrients for limited periods.
• Media : Diluted Knop's solution supplemented by glucose, some aminoacids,
Liebig's meat extract
• No sub-culture attempts
• 1922- Robbins and Maneval : Using subcultures, succeeded in
maintaining cultures of corm root for 20 weeks.
• Media : Yeast extract
Ten years later, White (1934a), working with tomato roots, obtained unlimited
cultures with Robbins' medium, also containing yeast extract.
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9. • 1925- F. Liabach Demonstrated application of embryo culture by
crossing Linum perenae with L. austriacum to get hybrid plants from
shriveled seeds.
• 1926- Went : Discovered Auxin – (Kogl: IAA )
• 1934- P R White :Established continuously growing root cultures of
tomato
On this medium he could maintain root cultures for 34 years by continuous sub
culturing (till his death at 1968)
Medium containing
1. Inorganic salts
2. Yeast extract
3. Sucrose
1. Pyridoxine
2. Thiamine
3. Nicotinic acid
Later yeast extract was replaced by
three components of vitamin B
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(Gautheret, 1983)

10. • Cultured cambium cells of some
tree species
• Medium : Knop’s solution with
Glucose, Cystein HCl, solidified
with agar
• Callus formation after 2 months
• After 6 months, growth was
ceased due to nutrient
deficiency
1930-34- R. J. Gautheret
 Salix caprea
 Acer psedoputamus
 Ulmus
1935 - Snow
• Demonstrated that IAA stimulated
cambial activity
P. R White – (1939)
• Made similar observation from
tumour tissues of the hybrid N.
glauca and N. longsdorfii
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Conti..

11. • 1939- Philip R. White, R. J. Gautheret and P. Nobecourt :
Independently announced the possibility of cultivating plant tissues
for unlimited periods.
Philip. R. White R. J. Gautheret Pierre Nobecourt
 P. R. White : Tumors tissue of hybrid Nicotiana glauca x Nicotiana langsdorfi
 R. J. Gautheret and P. Nobecourt : Carrot tissue
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Conti..

12. • 1942- P. R. White and A. C. Braun: Initiated studies on crown gall and
tumour formation in plants.
• 1944- F. Skoog: Started his work on organogenesis in tobacco callus.
Bud culture
• 1945- Loo: Obtained excellent cultures of Asparagus and dodder stem
tips.
• 1946- E.A. Ball: Father of micropropagation
Development of plantlets from sterile cultures of stem tips in
Tropaeolum and Lupinus.
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Conti..

13. Thanks to coconut milk : Discovery of
Cytokinines
• 1940- Blakeslee – Embryo culture of Datura hybrid in liquid medium
of coconut milk
• 1948 - Caplin and Steward: Applied coconut milk in the tissue
culture field for the first time- Strong proliferation of carrot tissue
• 1950- Morel: Obtained the indefinite growth of tissue from a
monocotyledon species, Amorphophalus rivieri using medium
enriched with coconut milk
What might be growth promoting factor in coconut milk?
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14. • 1951- Skoog and Tsui : The continued induction of cell division, callus
growth and bud formation in tobacco pith tissue cultures were
achieved by adding adenine and high concentrations of phosphate.
• 1952- F. C. Stward, S. M. Caplin and F. K. Miller Discovered the
synergistic action of 2, 4-D and coconut milk in a culture of potato
tissue.
• 1954- Muir, W.H. Hildebrandt A.C. and Riker. A.J: First suspension
cultures by transferring callus (Tagetes erecta and Tobacco) fragments
to agitated liquid medium
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Conti..

15. • 1958- Muir : Nurse culture technique
• 1955- Skoog, Miller, Okumura, Von Saltza and Strong: undertook
experiments with nucleic acid extracted from herring sperm; calf
thymus and yeast.
Isolated – Kinetin (6-furfurylaminopurine);
A derivative of adenine–From yeast extracts 
The first member of the cytokinin family
Folke A. Skoog
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• 1957- Skoog and Miller : Organogenesis (tobacco explant) was
controlled by a balance between auxin and kinetin, the former
introducing root initiation and the second bud organization.

16. Toshio Murashige
• 1963- Letham : Isolated Zeatin ; Coconut milk contain
Ribosylzeatin
• 1962- Murashige and Skoog (1962)- MS medium
• 1958- Jacob Reinert (Germany) and F. C. Steward (USA):
Somatic embryogenesis in suspension culture of carrot.
Steward and Reinert independently reported bipolar somatic
embryos in carrot 16

17. • 1960- E. C. Cocking : Protoplast isolation by enzymatic degradation
• 1960- L. Bergmann : Bergmann plating technique. This plating
involved mixing cells with warm sugar medium just prior to gelation in
petridish.
• 1964- S. Guha and P. Maheshwari: Haploid plant induction via anther
culture - Datura innoxia
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S. Guha mukharjee
P. Maheshwari
https://fellows.ias.ac.in/profile/v/FL1988026

18. • 1970- Kasha, K. J and Kao, K.N: Bulbosum technique in Hordeum
 Hordeum vulgare x Hordeum bulbosum  Haploid plants of H. vulgare
Elimination of bulbosum chromosome occurred.
• 1972- Withers L and Cocking E C: Laid foundation for the protoplast
fusion technique.
• 1972- Carlson and collaborators : Plants regenerated from fusion
between protoplast of N. glauca and N. langsdorffi
• 1973 – Melchers and co workers: Fusion of protoplast from tomato
and potato
• 1973- Inkgo Potrykus: First attempt on chloroplast and nucleus transfer
from Petunia hybrids into albino protoplasts of the same species.
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19. • 1974- Melchers, G. and Labib, G proposed hybrids resembling the sexual
hybrids by fusing protoplasts of two varieties of Nicotiana tabacum
• 1974- Murashige: Developed the concept of developmental stages in
cultures in vitro: Stage I: Explant establishment; Stage II: Multiplication of
propagule and Stage III: Rooting and hardening for planting into soil.
• 1981- Larkin P. J and Scowcroft W. R:
Developed the concept of somaclonal variation: A noval source of
variability from cell cultures for plant improvement
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20. • 1982- Krens, F.A., Molondijk, L. Williams G. J. and Schilperoort, R.A
developed poly ethylene glycol method for the direct delivery of DNA
into protoplasts.
• 1985- Gheysen G., Dahese P., Van Montaque M and Schell J
Developed very efficient gene transfer system using natural gene
transfer mechanism of Agrobacterium tumifaciens.
• 1986- Pirrie, A. and Power, J.B: Produced fertile, interspecific gameto-
somatic triploid hybrids of tobacco by fusing protoplasts of leaf (2n)
and pollen tetrad (n).
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(Thorpe, 2007)

21. • 1986- Kinsara A., Patnaik, S.N., Cocking, E.C. and Power, J.B
Produced somatic hybrids between Lycopersicon esculentum and L.
peruvianum.
• 1987- Terada R. Kyozuka J. Nishibayashi S and Shimamoto K
Regenerated plantlets form somatic hybrid cells of Oryza sativa, and
Echinochloa oryzicola.
• 1990- Milanova V and Zagorska N A: Succeeded in overcoming hybrid
incompatibility between Nicotiana africana and N. tabacum and
produced cytoplasmic male sterile plants by embryo culture.
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(Thorpe, 2007)

22. Conclusion
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23. References
• Gautheret, R. J. 1983. Plant tissue culture: A history. Bot. Mag. 96:
393-410.
• Thorpe, T. A. 2007. History of plant tissue culture. Mol. Biotechnol..
37: 169-80.
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24. THANK’s
to science