Plant tissue culture involves growing plant cells, tissues, organs, or whole plants in vitro on a nutrient medium under sterile conditions. It allows for mass propagation of plant materials, rapid plant breeding through selection of variants, and genetic manipulation. The key principles involve using plant hormones like auxin and cytokinin to induce cell differentiation and regeneration into whole plants. Advantages include rapid multiplication, disease elimination, genetic transformation, and conservation of endangered species.

1. INTRODUCTION TO
PLANT TISSUE CULTURE
TECHNOLOGY

2. PLANT TISSUE CULTURE ?
Any technique of growing cells/tissues/organs/seeds/
embryos or other plant parts in a controlled environment
on a nutrient medium
Cultural techniques for regeneration of functional
plants from different tissues/organs/cells/protoplasts
Also known as micropropagation
The growth or maintenance of plant cells, tissues,
organs or whole plants in vitro.
 the science of growing plant cells/tissues/organs
isolated from the mother plant on artificial media.

4. • Faster propagation  reduce the time
• Speed up the selection process for crop
improvement  save time
• Disease free plants
• Save space  reduce cost
• Pharmaceutical properties
• Conservation of endangered species
• Management of genetic resources
WHY?

5. WHY?
 Mass propagation/production
 Plant breeding
 selection of somaclonal variants
 mutation selection
 haploid/dihaploid production
 In-vitro hybridization – protoplast fusion
 Genetic manipulation
 Secondary product synthesis

6. HISTORY – Chronology of Important Development
 1902 (Gottlieb Haberlandt)
 culture isolated, differentiated cells from leaves of various
plants on salt solution (Knop’s) with sucrose.
 maintain cells alive for a period but not divided (lack of
plant hormones).
 continued cell division in explanted tissues on
artificial medium.
 1922 (Robbins & Kotte)  culture of isolated root tips.
 1934 (White)  culture of tomato roots (long period).
 culture tomato meristematic cells on medium containing
salts, yeast extract, sucrose, vitamin B (pyridoxine, thiamine,
nicotinic acid)

7.  1939 (Gautheret & Nobecourt)  1st long term
callus culture involving cambial tissues of carrot.
 1939 (White)  Callus culture of tobacco tumor
tissues from interspecific hybrid of Nicotina glaucum x
N.longsdorffi.
 1941 (Van Overbeek)  Discovery of nutritional
value
 1942 (White & Braun)  Experiments on crown-
gall and tumor formation in plants, growth of bacteria
free crown-gall tissues.
 1948 (Caplan & Steward)  Use of coconut milk +
2,4-D for proliferation of carrot and potato.
HISTORY-(cont.)

8.  1950 (Morel)  Culture of monocot tissues using
coconut milk.
 1953 (Muir)  Innoculation of callus in liquid medium.
Development of technique for single isolated cell culture.
 1953 (Tulecke)  Haploid culture from pollen
gymnosperm (Ginkgo).
 1955 (Miller et al.,)  Discovery of cytokinins
(kinetin).
 1955 (Ball)  Culture of gymnosperm tissues (Sequoia).
 1957 (Skoog & Miller)  Hypothesis that shoot and
root initiation in cultured callus is regulated by the
proportion of auzin and cytokinins.
HISTORY-(cont.)

9.  1960 (Cocking)  Enzymatic isolation and culture of
protoplast.
 1960 (Morel)  Development of shoot apex culture
technique.
 1964 (Morel)  Use of modified shoot apex
technique for orchid proportion.
 1966 (Guha & Maheshwari)  Cultured anthers
and pollen and produce haploid embryos.
 1974 (Nitsch)  Culture of microspores of Datura
and Nicotina to double up the chromosome number.
HISTORY-(cont.)

10.  1978 (Melchers)  Production of somatic hybrids
from attached to plasmid vectors into naked plant
protoplast.
 1983 (Barton et al.,)  Insertion of foreign genes
attached to plasmid vectors into naked plant protoplast.
 1983 (Chilton)  Production of transformed tobacco
plants following single cell transformation or gene
insertion.
HISTORY-(cont.)

11. BASIS
• Plant hormones
 Auxin
 Cytokinin
 The ratio can determine plant growth and
development
 Auxin > Cytokinin
 Auxin < Cytokinin
 auxin = Cytokinin

12.  Totipotency of plant cells
 ability to develop into whole plants/plant
organs in vitro
 not all cells are totipotent
 Differentiated cells can be dedifferentiated
 callus
 callus can be redifferentiated back to
somatic embryo which later regenerate the
entire plant
BASIS/PRINCIPLE

13.  Growth Media
 minerals, C source, hormones, etc.
 Environmental
 light, T, photoperiod, etc.
 Explant Source
 age, type of organ, etc.
 Genetics
 diff species or genotype have diFf
response
FACTORS

14. • Adventitious - Developing from unusual points of origin, such as
shoot or root tissues, from callus or embryos, from sources other
than zygotes.
• Agar - a polysaccharide powder derived from algae used to
solidify a medium.
• Aseptic - Free of microorganisms.
• Aseptic Technique - Procedures used to prevent the introduction
of microorganisms into cultures.
• Autoclave - An equipment that provides heat under high steam
pressure for sterilization.
• Axillary bud/shoot - bud/shoot arises from the axil of leaves or
normal origin.
• Callus - An unorganized, proliferate mass of differentiated plant
cells, a natural response of wound.
TERMINOLOGY

15. • Chemically Defined Medium - A nutritive solution for culturing
cells in which each component is specifiable and ideally of known
chemical structure.
• Clone - Plants produced asexually from a single source plant.
• Clonal Propagation - Asexual reproduction of plants that are
considered to be genetically uniform and originated from a single
individual or explant.
• Contamination - Being infested with unwanted microorganisms.
• Culture - plant growing in vitro.
• Differentiated - Cells that maintain, in culture, all or much of the
specialized structure and function typical of the cell type in vivo.
Modifications of new cells to form tissues/organs with a specific
function.
• Epigenetic changes - persistent changes in phenotype that involve
the expression of particular genes.
TERMINOLOGY

16. • Explant - Tissue taken from its original site and transferred to an
artificial medium for growth or maintenance.
• Hormones - Growth regulators, generally synthetic in occurrence,
that strongly affects growth (i.e. cytokinins, auxins, and
gibberellins).
• Internode - The space between two nodes on a stem.
• In vitro - isolated from living organism and artificially maintained.
• Media - Plural of medium.
• Medium - A nutritive solution, solid or liquid, for culturing cells.
• Micropropagation - In vitro clonal propagation of plants from
shoot tips or nodal explants, usually with an accelerated
proliferation of shoots during subcultures.
• Organ - a distinct and visibly differentiated part of an organism.
TERMINOLOGY

17. • Organogenesis - refers to forming organs, in particular both
roots and shoots from callus cells.
• Node - A part of the plant stem from which a leaf, shoot or
flower originates.
• Passage - Transfer or transplantation of cells/tissues with or
without dilution/division, form one culture vessel to another.
• Passage Number - The number of times the cells or tissues in
culture have been subcultured or passaged.
• Pathogen - A disease-causing organism.
• Pathogenic - Capable of causing a disease.
• Petiole - A leaf stalk; the portion of the plant that attaches the
leaf blade to the node of the stem.
• Plantlets - small complete plants that were produced via tissue
culture.
TERMINOLOGY

18. • Propagules - tissue that is divided and used for further
multiplication.
• Regeneration - a morphogenetic response to a stimulus that
results in the products of organs, embryos, or whole plants.
• Somaclonal Variation - Phenotypic variation, either genetic or
epigenetic in origin, displayed among somaclones.
• Somaclones - Plants derived from any form of cell culture
involving the use of somatic plant cells.
• Somatic - cells/tissues that are vegetative and have the
complete genetic material of the organism (plant).
• Sterile - (A) Without life. (B) Inability of an organism to
produce functional gametes. (C) A culture that is free of viable
microorganisms.
• Sterile Techniques - The practice of working with cultures in
an environment free from microorganisms.
TERMINOLOGY

19. • Subculture - See “Passage”. The process by which the tissue or
explant is first subdivide, then transferred into fresh culture
medium.
• Tissue Culture - The maintenance or growth of tissue, in vitro,
in a way that may allow differentiation and preservation of
their function.
• Totipotency - A cell characteristic in which the potential for
forming all the cell types in the adult organism are retained.
• Undifferentiated - With plant cells, existing in a state of cell
development characterized by isodiametric cell shape, very
little or no vacuole, a large nucleus, and exemplified by cells
comprising an apical meristem or embryo.
TERMINOLOGY

20. ADVANTAGES & APPLICATION
 Can grow plant cells in liquid culture on a large
scale
 Shortens the time taken to achieve uniform
homozygous lines and varieties
 Protoplast isolation and somatic fusion of
distantly related species increases the
possibility for the transfer and expression of
novel variation in domestic crops
 Increases the potential number of individuals
in a screening program

21.  Allows the production of large numbers of
uniform individuals of species from limited
starting material
 Enables very specific information to be
introduced into single cells which can then be
regenerated
 Overcomes seasonal restriction for
germination or cultivation
 Enable conservation, preservation,
international exchange of materials
ADVANTAGES

22. DISADVANTAGES?