BSc III.pdf Plant tissue culture study material

BSc III.pdf Plant tissue culture study material

1. PLANT TISSUE CULTURE TECHNIQUES —
Dr. Rajesh Kumar
Assistant Professor of Botany, MGGAC, Mahe

2. WHAT IT IS?
•In vitropropagation? Or
•Micropropagation?Or
•In vitroculture?
“… THE ASEPTİC CULTURE OF PLANT…”
Implies-regeneration
-multiplication

3. •True-to-type clones
•A single explant can be multiplied into several thousand
•Year-round production
•Rare and endangered plants can be cloned safely
•To produce virus free plants
•Long-term germplasm storage with ‘tissue banks’
•Plant cultures easier to export than are soil-grown plants
•Production of difficult-to-propagate species
•Worldwideindustrymultibillion Euros
IMPORTANCE OF PLANT TISSUE CULTURE TECHNIQUES

4. FUNDAMENTAL AB ILITIES OF PLANTS
HOW CAN A PLANT CELL OR TISSUE DEVELOP?
Totipotency
Dedifferentiation
Competency
Therefore, tissue can be regenerated from
explants such as cotyledons, hypocotyls,
leaf, ovary, protoplast, roots, anthers,
etc.

5. WHAT’S THE BACKGROUND
1902–Haberlandt –The Concept
1920s –Knudson –Simple Orchid Germination–First commercial use
1930s –Thimann & Went –Auxin
1930s –White/Gautheret/Nobecourt –Root Cultures
1950s –Skoog’s group –Cytokinins,
–The discovery of the structure of DNA by Crick and Watson
1960s –Morel-Orchid micropropagation, thermotherapy
1970’s –Genetic engineering took off
1990s –by Calgene –genetically engineeredpotatoes
Gottleib Haberlandt

6. WHAT IS NEEDED?
•Appropriate tissue
•A suitable growth medium
•Aseptic (sterile) conditions
•Growth regulators:The ratio of auxins and cytokinins
•Frequent subculturing

7. TYPESOF PLANT TISSUE CULTURE TECHNIQUES
1.Culture of intact plants (Seed orchid culture)
2.Embryo culture (embryo rescue)
3.Organ culture:Micropropagation
A.Organogenesis in solid or semi solid medium
1. Meristem and shoot tip culture
2. Bud culture
3.Root culture
4. Leaf culture
5. Anther culture
B. Somatic embryogenesis
C. Organogenesis and somatic embryogenesis in bioreactors
D. In vitro micrografting
E. Thin cell layer technology (TCLs)
F. Photoautotrophic culture
4. Callus culture
5. Cell suspension and single cell culture
6. Protoplast culture, somatic hybridization

8. Culturing (micropropagating)
Plant Tissue -the steps
•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

9. ORGANOGENESIS OF PISTACHIO
Culture initiation
Shoot proliferationRootingHardened plantlets
Fresh apical tip (a)
or nodal bud (b)1 2 3
6 5 4
Lignified shoots

10. ADVENTITIOUS ORGANOGENESIS IN PISTACHIO
Adventitious Buds
Regeneration of Plantlets
from Single Leaflet2
2
Elongation and
3
4
2
6 5
1 2 3
4

11. SOMATIC EMBRYOGENESIS IN PISTACHIO
Isolated kernels
Swollen SEs Maturation of SE
Ebryogenic tissue Ebryogenic tissues
in liquid medium
Development of SEs
Germinated SEs Acclimatised somatic
seedlings
Immature fruits
Isolated SEs for
germination
6 months after transplanting
somatic seedlings
Ebryogenic tissues

12. MICROPROPAGATION IN BIOREACTORS
Bioreactor;tissue culture containersAutomated culture system The design of separate compartments

13. WHAT IS MICROGRAFTING?

14. The thin cell layer (TCL) system consists of explants of small size excised from different
plant organs either longitudinally (lTCL) or transversally (tTCL)

15. PHOTOAUTOTROPH IC CULTURE

16. Through micropropagation, it is now possible to provide clean and uniform planting materials in
plantations for several plant species such as oil palm, plantain, pine, banana, abaca, date, rubber
tree; field crops –eggplant, jojoba, pineapple, tomato; root crops –cassava, yam, sweet potato; and
many ornamental plants such as orchids and anthuriums (Alfonso, A. 2007; Singh et al. 2011).
Bioreactor cultures are being established in several commercial laboratories for micropropagation
of
ferns, spathiphylum, philodendron, banana, potato, lilies, poinsettia, sugar-cane, and some forest tree
species such as eucalyptus, poplar, and early stages of conifer somatic embryos (Aitkin-Christie, 1991;
Mehrotra et al 2007;Gross and Levin, 1999; Cervelli and Senaratna 1995).And plant products,
pharmaceuticals, food ingredients and cosmetics(Perulllini et al., 2007; Vongpaseuth and Roberts
2007; Pavlov et al. 2007)
Micrografted seedlings are commercialised to avoid the serious crop loss caused by infection of soil-
borne diseases for fruit trees and several vegetables(Navarro et al., 1975;Navarro 1981;Jung-
Myung Lee et al. 2010).
Transverse thin layer section technology may be ideal for large scale micropropagation of
ornamental plants(Jain et al. 1998).
Photoautotrophic flow-through systems for enhanced micropropagationfor Gerbera; Hypericum,
Myrtus communis, Momordica grosvenori, Eucalyptus (Nguyen and Kozai 2005; Xiao et al.2011)
The development of transgenic methods and the growth of agricultural biotechnology started
during the 1980s and the global biotech crophas increased hundred million of hectares area
Palmer et al. 2005; Thomas et al. 2003
Efficient doubled haploid technology enables breeders to reduce the time and the cost of new
cultivar development relative to conventional breeding practices.
APPLICATIONS OF MICROPROPAGATION

17. •Adaptation of tissue culture technology to more
species
•Fast and mass propagation of transformed plants
with“designer genes”
•Chloroplast transformation methods
•Efficient, computer-controlled flow-through
systems to cut down the labor cost
•Mass production of plant constituents
•New technologies
WHAT’S IN THEFUTURE