Gottlieb Haberlandt is considered the father of plant tissue culture, as he first developed the concept of culturing isolated plant cells in artificial conditions in 1902. Though his experiments failed, he established the concept of totipotency. In the 1930s-1940s, Gautheret and White established tissue culture techniques and nutrient media that are still used today. Important milestones included the discoveries of auxins, kinetin, protoplast isolation techniques, and the development of the Murashige and Skoog medium in 1962. Modern plant biotechnology resulted from over a century of basic research on topics like tissue culture, organogenesis regulation, and bacterial-plant interactions combined with technological innovations in cell and
2. Plant Tissue Culture
Tissue culture is the in vitro aseptic culture of cells, tissues, organs or
whole plant under controlled nutritional and environmental
conditions. often to produce the clones of plants. The resultant clones
are true-to type of the selected genotype. The controlled conditions
provide the culture an environment conducive for their growth and
multiplication. These conditions include proper supply of nutrients,
pH medium, adequate temperature and proper gaseous and liquid
environment.
3. PTC- How It started
• During the 1800s, the cell theory, which states that the cell is the
basic structural unit of was very quick to gain acceptance.
• However, the second portion of the
cell theory states that these structural units are distinct and
potentially totipotent physiological and developmental units, failed to
gain universal pass acceptance.
• The skepticism associated with the latter part was because of the
inability of scientists such as Schleiden and Schwann to demonstrate
totipotency in their laboratories
4. Historical Developments: PTC
• It was in 1902 that the well-known German plant physiologist,
Gottlieb Haberlandt (1854-1945), attempted to cultivate plant tissue
culture cells in vitro.
• He clearly stated the desirability of culturing the isolated vegetative
cells of higher some plants.
• He stated: "To my knowledge, no systematically organized attempts to
culture isolated vegetative cells from higher plants in simple nutrient
solutions have been made.
5. Historical Developments: PTC
• Yet the results of such culture experiments should give some
interesting insight to the properties and potentialities that the cell, as
an elementary organism, possesses. Moreover, it would provide
information about the interrelationships and complementary
influences to which cells within a multicellular whole organism are
exposed.
6. Historical Developments: PTC
• He experimented with isolated photosynthetic leaf cells and other
functionally differented cells and was unsuccessful, but nevertheless
he predicted that one could successfully cultivate artificial embryos
from vegetative cells. He, thus, clearly established the concept of
totipotency, and further indicated that the technique of cultivating
isolated plant cells in nutrient solution permits the investigation of
important problems from a new experimental approach
7. Historical Developments: PTC
On the basis of that address
and his pioneering
experimentation before and
later, Gottlieb Haberlandt
(1854-1945), is justifiably
recognized as the father of
plant tissue culture.
8. Historical Developments: PTC
• In 1902, a German Botanist Gottlieb Haberlandt developed the
concept of culture of isolated cells of Tradescantia in artificial
condition. Though his experiment failed to induce the cells to divide.
• He did not succeed because by that time even auxin was not
discovered. But he lent a foundation to plant physiology.
• He described the cultivation of mesophyll cells of Lamium purpureum
and Eichhornia crassipes, epidermal cells of Ornithogalum and hair
cells of Pumonaria
11. Historical Developments: PTC
• Cell survived for 3-4 weeks. Due to this endeavour, Haberlandt is
regarded as the father of tissue culture. Most importantly he
suggested the concept of totipotency.
• From 1902 to 1930 attempts were made for organ culture.
• Hannig (1904) isolated embryos of some crucifers and successfully
grew on mineral salts and sugar solutions.
• Simon (1908) successfully regenerated a bulky callus, buds, roots
from a poplar tress on the surface of medium containing IAA which
proliferated cell division.
12. Historical Developments: PTC
The two important discoveries made in the mid 1930s which gave a big
push to the development of plant tissue culture technique were:
• (a) identification of auxin as a natural growth regulator, and
• (b) recognition of the importance of B-vitamins in plant growth.
13. Historical Developments: PTC
In 1934, Gautheret had cultured cambium cells of some tree species
(Salix capraea, Populus nigra) on Knop's solution containing glucose
and cysteine hydrochloride and recorded that they proliferated for a
few months.
Salix capraea
14. Contributions of Gautheret
• The first true plant tissue cultures were obtained by Gautheret from
cambial tissue of Acer pseudoplatanus.
• He also obtained success with similar explants of Ulmus campestre,
Robinia pseudoacacia and Salix capraea using agar-solidified medium
of Knop’s solution, glucose and cysteine hydrochloride.
• The first continuously growing tissue cultures from carrot root
cambium were established by Gautheret in 1939.
15. Philip Rodney White (1901–1968)
• White (1939) reported the
establishment of similar
cultures from tumour tissue
of the hybrid Nicotiana
glauca x N. langsdorffii.
• Then the possibility for
cultivation of plant tissues for
unlimited period was
announced simultaneously
by P.R. White (1939) and R.J.
Gautheret (1939).
16. Philip Rodney White (1901–1968)
• Unlimited Growth of Cultured Root Tips
• Unlimited Growth of Callus Tissues
• Autonomous Growth of Bacteria-Free Secondary Crown Gall Tumors
• Development of Chemically Defined White's Nutrient Solution
• Founding of the International Association for Plant Biotechnology
17. Historical Developments: PTC
• Gautheret and White during 1930-40 were responsible for
establishing the media composition we use today
• Subsequent detailed work by Raghavan and Torrey (1963), Norstog
(1965) and others led to the development of synthetic media for the
culture of younger embryos
• During 1940 to 1970, suitable nutrient media were developed for
culture of plant cells, tissue, protoplasts, anthers, roots tips and
embryos.
• in vitro morphogenesis (i.e. regeneration of complete plant from
cultured tissue) of plants was always successfully done.
18. Historical Developments: PTC
• In 1957, Skoog and Miller put forth the concept of hormonal control
of organ formation
• Murashige was instrumental in giving the techniques of in vitro
culture a status of a viable practical approach to propagation of
horticultural species. He worked extensively for the popularization of
the technique by developing standard methods for in vitro
propagation of several species ranging from ferns, to foliage, flower
and fruit plants.
19. Historical Developments: PTC
• In 1959, discovery of kinetin promoted by F. Skoog along with C.O.
Miller and co-workers and demonstration of induction of
regeneration of shoots in tobacco callus paved the way for
multiplication of plant by tissue culture.
• In 1960s, E. Cooking for the first time developed a method for
isolation of protoplasts in large quantities using the fungal enzyme
obtained from Myrothecieum sp.
20. Historical Developments: PTC
• In 1960 Jones et al. designed a microculture method for growing
single cells in hanging drops in a conditioned medium
• The first plant from a matured plant cell was regenerated by Braun in
1959.
21. Murashige and Skoog medium
Murashige and Skoog medium (MSO or MS0 (MS-
zero)) is a plant growth medium used in the
laboratories for cultivation of plant cell culture. MSO
was invented by plant scientists Toshio
Murashige and Folke K. Skoog in 1962 during
Murashige's search for a new plant growth regulator.
A number behind the letters MS is used to indicate
the sucrose concentration of the medium. For
example, MS0 contains no sucrose and MS20
contains 20 g/l sucrose. Along with its modifications,
it is the most commonly used medium in plant tissue
culture experiments in the laboratory
22. Murashige and Skoog medium
• As Skoog's doctoral student, Murashige originally
set out to find an as-yet undiscovered growth
hormone present in tobacco juice. No such
component was discovered; instead, analysis of
juiced tobacco and ashed tobacco revealed
higher concentrations of specific minerals in
plant tissues than were previously known. A
series of experiments demonstrated that varying
the levels of these nutrients enhanced growth
substantially over existing formulations. It was
determined that nitrogen in particular enhanced
growth of tobacco in tissue culture.
23. Historical Developments: PTC : Indian Story
In India, work on tissue culture was started during
mid 1950s at the Department of Botany
(University of Delhi) by Panchanan Maheshwari
who is regarded as father of embryology in India.
During 1960s the Botany School at the University
of Delhi, led by P. Maheshwari, became actively
engaged with in vitro culture of reproductive
organs of flowering plants.
• Kanta, 1960 developed the technique
'intraovarian pollination’ and 'test-tube
fertilization'
24. Historical Developments: PTC : Indian Story
• Different tissue culture methodologies were involved for
morphogenic studies involving ovary, embryo, endosperm, ovules,
etc.
• At the University of Delhi, Sipra Guha Mukherjee and S.C.
Maheshwari for the first time developed the haploid through anther
and pollen cultures.
• Haploid plants from pollen grains were first produced by Maheswari
and Guha in 1964 by culturing anothers of Datura.
26. CONCLUDING REMARKS
• Modern plant biotechnology, defined as the genetic modification
of plants, resulted from a century-long combination of basic
research findings and technological innovations. The basic
scientific findings that underlay this include in vitro tissue culture,
auxin/cytokinin regulation of organogenesis, single cell culture,
discovery of cellular totipotency, the bacterial cause of crown
gall disease, opines as markers of transformed cells,
transfer of virulence between Agrobacterium strains, T-DNA, the
genes that determine tumor morphology (tms1, tms2, and tmr),
disarmed plasmids, and regeneration of transformed cells.
• The technological innovations include aseptic tissue/cell culture,
hanging drop culture, micropipettes, nurse cultures, binary plant
vectors, and gene gun transformation.
