Plant Tissue Culture

1. Plant Tissue Culture

2. What is Plant Tissue Culture?Tissue culture is the culture and maintenance of plant cells, tissues or organs (explants) in sterile, nutritionally (synthetic media) and environmentally (controlled) supportive conditions (invitro).

3. What conditions do plant cells need to multiply in vitro? • Freedom from competition • Nutrients and removal of waste products • A controlled environment

4. Why Tissue Culture?

5. Seed culture • Increasing efficiency of germination of seeds that are difficult to germinate in vivo • Precocious germination by application of plant growth regulators • Production of clean seedlings for explants or meristem culture

6. Embryo culture • Overcoming embryo abortion due to incompatibility barriers • Overcoming seed dormancy and self-sterility of seeds • Embryo rescue in distant (interspecific or intergeneric) hybridization where endosperm development is poor • Shortening of breeding cycle

7. Ovary or ovule culture • Production of haploid plants • A common explant for the initiation of somatic embryogenic cultures • Overcoming abortion of embryos of wide hybrids at very early stages of development due to incompatibility barriers • In vitro fertilization for the production of distant hybrids avoiding style and stigmatic incompatibility that inhibits pollen germination and pollen tube growth

8. Anther and microspore culture • Production of haploid plants • Production of homozygous diploid lines through chromosome doubling, thus reducing the time required to produce inbred lines • Uncovering mutations or recessive phenotypes

9. In vitro pollination • Production of hybrids difficult to produce by embryo rescue

10. Organ culture • Any plant organ can serve as an explant to initiate cultures

11. Shoot apical meristem culture • Production of virus free germplasm • Mass production of desirable genotypes • Facilitation of exchange between locations (production of clean material) • Cryopreservation (cold storage) or in vitro conservation of germplasm

12. Somatic embryogenesis • One major path of regeneration • Mass multiplication • Production of artificial seeds • As source material for embryogenic protoplasts • Amenable to mechanization and for bioreactors

13. Organogenesis • One major path of regeneration • Mass multiplication • Conservation of germplasm at either normal or sub-zero temperatures

14. Enhanced axillary budding • Micropropagation

15. Callus Cultures • In some instances it is necessary to go through a callus phase prior to regeneration via somatic embryogenesis or organogenesis • For generation of useful somaclonal variants (genetic or epigenetic) • As a source of protoplasts and suspension cultures • For production of metabolites • Used in in vitro selection

16. In vitro mutagenesis • Induction of polyploidy • Introduction of genetic variability

17. Protoplast isolation, culture and fusion • Combining genomes to produce somatic hybrids, asymmetric hybrids or cybrids • Production of organelle recombinants • Transfer of cytoplasmic male sterility

18. In vitro flowering • This can be done in some instances but I am not sure there are any practical applications

19. Micrografting • Overcoming graft incompatibility • Rapid mass propagation of elite scions by grafting onto rootstocks that have desirable traits like resistance to soil-borne pathogens and diseases • To allow survival of difficult to root shoots • Development of virus free plants

20. Genetic transformation • Many different explants can be used, depending on the plant species and its favored method of regeneration as well as the method of transformation • Introduction of foreign DNA to generate novel (and typically desirable) genetic combinations • Used to study the function of genes

21. Development of pre-existing structures De novo formation based on totipotentiality Axillary bud Adventitious bud

22. Tuberization Flowering in vitro

23. Ovary development Rhizogenesis

24. Grafting in vitro Bulbil development

25. Embryogenesis

26. Somatic embiryogenesis in cotton

27. History of plant tissue culture • TOTİPOTENSİ Cell theory • SCHLEIDEN 1838 in plants, • SCHWANN 1839 in plants and animals • Among the lower plants any cell can be separated from the plant and continue to grow. Thus, entire plants may consist of cells whose capacity for independent life can be clearly demonstrated.

28. Haberland, 1902 (first aseptically culture attemp in a nutrient solution

29. White,1934 First root culture

30. The in vitro cultivation of plant tissues for indefinite periods of time was simultaneously achieved by WHITE working with a Nicotiana hybrid and GAUTHERET and NOBECOURT working with carrot Daucus carota in 1939. This achievement in both cases was due to a fortunate choice of plant material as both are considered to be relatively easy to culture now in comparison with many other species but this in no way negates the magnitude of their efforts.

31. Gautheret, first callus culture

32. Skoog, 1954

33. Murashige ve Skoog medium Murashige

34. Maheswari, 1960 first anther culture

35. Nitsch, 1974 microspore culture

36. Cocking, 1960 Protoplast culture

37. Morel, 1960 micropropagationMelchers, 1978 protoplast fusion Pomato

38. Nickell, Sekondary metabolite production

39. Plant tissue culture studies in TURKEY • First at Agricultural Resarch Institions and Universities with micropropagation studies • Aegean and Ankara Universities and Bornova Agricultural Resarch Instition are pioneering • Today more than 30 Universities, Rsearch Institions, TÜBİTAK and private sector has tissue culture facilities.

40. Growth chambres * * laminar flows *