1. The Three Pillars of Yield
BREEDING
Strategically breed plants to create new, more robust seeds that perform better – and longer – in the field.
AGRONOMICS
Use precision ag, planting density, plant health protection, and conservation tillage to make acres more productive.
BIOTECHNOLOGY
Supplement breeding advancements by adding special beneficial genes to the plant.
ALL THREE ARE CRITICAL IN DELIVERING YIELD TODAY – AND TOMORROW 1

2. In Vitro Culture
The culture of organized living material cell under artificial condition in tubes, glasses, dishes
The process of growing organized living materials artificially in the laboratory
The culture and maintenance of organized living materials, in sterile, nutritionally (synthetic media) and environmentally (controlled) supportive conditions

3. Organized living material
Heterogeneous (organs)
Homogeneous (tissues)

4. Tissue
Hooke (1665) and Leeuwenhoek (1677) using microscope discovered tissues were made of cells

5. The cells of a particular tissue had a common structure.

6. Cell Taken to its simplest form A plasma membrane…
Surrounding cytoplasm…
Containing hereditary material.

7. Level of complexity Organism Organ-system Organ Tissue Cell
Xavier Bichat ( ): An organ is composed of different tissues
Several organs can be grouped together as an organ system (e.g. the digestive system)
Organism
Organ-system
Organ
Tissue
Cell

8. Level of complexity
Purkinje (1835) Observed a fertilised hen's egg (a single cell) could develop into an embryo (many specialised cells in a compact mass)
C19th botanists showed that plant tissues consist of many different types of cells.

9. THE CELL THEORY
Schleiden (1838) & Schwann (1839) “The cell is the basic unit of living tissue”
The cell is an autonomous unit (“a citizen”) grouped together to form an organism (“the society”).

10. « Omnis cellula e cellula »
Virchow (1858) “all cells come from pre-existing cells”

11. Cell culture
The process by which prokaryotic, eukaryotic or plant cells are grown under controlled conditions.
Cell culture was first successfully undertaken in animal cell by Ross Harrison (1907) and in plant cells by Haberlandt (1902)

12. ORGANISMAL THEORY Reichert (a morphologist):
An organism has a structured plan
Image Credit Frog embryo fate map

13. Cells are connected in an organism sometimes by cytoplasmic bridges
Plasmodesmata
Strasberger (a cytologist):
Cells are connected in an organism sometimes by cytoplasmic bridges

14. Some organisms do not have cellular compartments
Acellular organisms
Some organisms do not have cellular compartments

15. Unicellular organisms
Some organisms only consist of a single cell
But these do usually have the cell components (nucleus, membrane, etc)

16. Tissue culture
In vitro cultivation of organs, tissues & cells at defined temperature using an incubator & supplemented with a medium containing cell nutrients & growth factors
Cells can be cultured away from a body
But this often requires elaborate support systems (technology)

17. Organized living material
Animal Cells
Plant Cells
Plant Tissue Culture

18. Plant Tissue Culture
“the aseptic culture of plant protoplasts, cells, tissues or organs under conditions which lead to cell multiplication or regeneration of organs or whole plants “

19. Totipotency … living plant cells can re-differentiate
…. each living cell has a complete genetic blueprint and therefore has the potential to develop into an entire plant.
… cells differentiate
… unlike animal cells, living plant cells can de-differentiate and then re-differentiate to form different cell types
… living plant cells can re-differentiate

20. Early Plant Cell Culture
…Haberlandt .. early 1900’s … proposed concept of totipotency … cells cultured under right conditions
Callus cultured from tree cambium Gautheret et al. (1930).
… cells kept alive but did not develop

21. Early Plant Tissue Culture
- dependent on discovery of “growth regulators”
Cell enlargement … role of auxins
Cell division ... role of cytokinins
Regeneration from tobacco pith (Skoog and Miller) … interaction of auxin and cytokinin gives differentiation.

22. Carrot plants from root cells
(Stewart, 1964)

23. Tobacco plants from single cells (Vasil & Hilderbrandt, 1965)

24. Requirement
Appropriate tissue (some tissues culture better than others)
A suitable growth medium containing energy sources and inorganic salts to supply cell growth needs. This can be liquid or semisolid
Aseptic (sterile) conditions, as microorganisms grow much more quickly than plant tissue and can over run a culture
Growth regulators, both auxins & cytokinins.
Frequent sub-culturing to ensure adequate nutrition and to avoid the build up of waste metabolites

25. Course Outline Introduction to in vitro culture
Determining factors in plant tissue culture
Micropropagation
Pattern of development
Micropropation enterprice