Plant tissue culture involves growing plant cells, tissues or organs in sterile conditions on nutrient media. Some key points: - Callus culture involves growing an unorganized cell mass (callus) from explants on auxin-cytokinin media, which can then be used for micropropagation. - Cell suspension culture breaks up callus into single cells that grow in liquid media, allowing for easier scaling up than callus. - Micropropagation is the process of rapidly multiplying plant materials like shoots, roots or embryos in culture to produce many clonal copies of plant materials like orchids or strawberries. - Protoplast isolation and culture allows the transfer of genes directly into plant cells without the

1. PLANT TISSUE
CULTURE
PRESENTED BY: MOHD NADEEM
M.Sc. BIOCHEMISTRY 3rd SEMESTER

2. Definition
the culture of plant seeds, organs,
tissues, cells, or protoplasts on
nutrient media under sterile
conditions.

3. HISTORY

4. STERILIZATION METHODS

9. Callus
 Equimolar amounts of auxin and cytokinin
stimulate cell division. Leads to a mass
proliferation of an unorganised mass of cells
called a callus.
 Requirement for support ensures that scale-
up is limited (Ginseng saponins successfully
produced in this way).

10. Cell suspension culture
 When callus pieces are agitated in a liquid
medium, they tend to break up.
 Suspensions are much easier to bulk up than
callus since there is no manual transfer or
solid support.

13. Types of In Vitro Culture
 Culture of intact plants (seed and seedling
culture)
 Embryo culture (immature embryo culture)
 Organ culture
1. shoot tip culture
2. root culture
3. leaf culture
4. anther culture
 Callus culture
 Cell suspension culture
 Protoplast culture

16. Micropropagation
 Embryogenesis
 Direct embryogenesis
 Indirect embryogenesis
 Organogenesis
 Organogenesis via callus formation
 Direct adventitious organ formation
 Microcutting
 Meristem and shoot tip culture
 Bud culture

17. Steps of Micropropagation
 Stage 0 – Selection & preparation of the mother plant
 sterilization of the plant tissue takes place
 Stage I - Initiation of culture
 explant placed into growth media
 Stage II - Multiplication
 explant transferred to shoot media; shoots can be constantly
divided
 Stage III - Rooting
 explant transferred to root media
 Stage IV - Transfer to soil
 explant returned to soil; hardened off

18. Features of Micropropagation
 Clonal reproduction
 Way of maintaining heterozygozity
 Multiplication Stage can be recycled many times
to produce an unlimited number of clones
 Routinely used commercially for many ornamental
species, some vegetatively propagated crops
 Easy to manipulate production cycles
 Not limited by field seasons/environmental influences
 Disease-free plants can be produced
 Has been used to eliminate viruses from donor plants

19. Embryo Culture
 Embryo culture developed from the need to rescue embryos
(embryo rescue) from wide crosses where fertilization occurred,
but embryo development did not occur
 These techniques have been further developed for the production
of plants from embryos developed by non-sexual methods (haploid
production discussed later)

21. Anther/Microspore Culture Factors
 Genotype
 As with all tissue culture techniques
 Growth of mother plant
 Usually requires optimum growing conditions
 Correct stage of pollen development
 Need to be able to switch pollen development from gametogenesis to
embryogenesis
 Pretreatment of anthers
 Cold or heat have both been effective
 Culture media
 Additives, Agar vs. ‘Floating’

22. PROTOPLAST ISOLATION

29. DIRECT GENE TRANSFER