Plant tissue culture techniques tools in plant micropropagation
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Plant tIssue culture technIques — Tools In plant mIcropropagatIon. Ahmet Onay , Department of Biology, Faculty of Science, University of Dicle, 21280 Diyarbakir, Turkey. EUROPEAN BIOTECHNOLOGY CONGRESS September 28 – October 1 , 20 11 İstanbul – TURKEY. W HAT IT IS?.

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Plant tIssue culture technIques — Tools In plant mIcropropagatIon

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Plant tissue culture techniques tools in plant micropropagation

PlanttIssueculturetechnIques—ToolsInplantmIcropropagatIon

Ahmet Onay, Department of Biology, Faculty of Science, University of Dicle, 21280 Diyarbakir, Turkey

EUROPEAN BIOTECHNOLOGY CONGRESS

September 28–October 1, 2011

İstanbul –TURKEY


W hat it is

WHAT IT IS?

  • In vitro propagation? Or

  • Micropropagation?Or

  • In vitro culture?

    “… THE ASEPTİC CULTURE OF PLANT…”

    Implies- regeneration

    - multiplication


Plant tissue culture techniques tools in plant micropropagation

IMPORTANCE OF PLANT TISSUE CULTURE TECHNIQUES

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

Worldwideindustry multibillion Euros


Fundamental ab i l i t i es of plants how can a plant cell or tissue develop

FUNDAMENTAL ABILITIES OF PLANTSHOW 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.


What s the background

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


What i s needed

WHAT IS NEEDED?

  • Appropriate tissue

  • A suitable growth medium

  • Aseptic (sterile) conditions

  • Growth regulators:The ratio of auxins and cytokinins

  • Frequent subculturing


Type s of plant tissue culture techniques

TYPES OF PLANT TISSUE CULTURE TECHNIQUES

  • Culture of intact plants (Seed orchid culture)

  • Embryo culture (embryo rescue)

  • 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


Culturing micropropagating plant tissue the steps

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


Organogenesis of pistachio

ORGANOGENESIS OF PISTACHIO

Fresh apical tip (a)

or nodal bud (b)

1

2

3

Culture initiation

Lignified shoots

Hardened plantlets

6

5

4

Rooting

Shoot proliferation


Plant tissue culture techniques tools in plant micropropagation

1

2

3

5

4

6

ADVENTITIOUS ORGANOGENESIS IN PISTACHIO

Adventitious Buds

2

Elongation and

2

2

3

from Single Leaflet

Regeneration of

Plantlets

4


Somatic embryogenesis in pistachio

Isolated kernels

Immature fruits

Ebryogenic tissue

Ebryogenic tissues

in liquid medium

Maturation of SE

Ebryogenic tissues

Development of SEs

Swollen SEs

Isolated SEs for germination

Germinated SEs

Acclimatised somatic seedlings

6 months after transplanting somatic seedlings

SOMATIC EMBRYOGENESIS IN PISTACHIO


Micropropagation in bioreactors

Bioreactor;tissue culture containers

Automated culture system

The design of separate compartments

MICROPROPAGATION IN BIOREACTORS


Plant tissue culture techniques tools in plant micropropagation

WHAT IS MICROGRAFTING?


Plant tissue culture techniques tools in plant micropropagation

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


Photoautotroph i c culture

PHOTOAUTOTROPHIC CULTURE


Plant tissue culture techniques tools in plant micropropagation

APPLICATIONS OF MICROPROPAGATION

  • 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 micropropagation for 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.


Plant tissue culture techniques tools in plant micropropagation

WHAT’S IN THEFUTURE

  • 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


Acknowledgements

ACKNOWLEDGEMENTS

All photos in this talk were taken by Ahmet Onay and Engin Tilkat, except the ones ontheslayt 12 and 15 which was obtainedfromthe WEB pages.

TUBITAK TOVAG - 3355 has grantedpart of thesestudies…

And the technical assistance of allco-authorsaregratefully acknowledged.

Ahmet Onay, Department of Biology, Faculty of Science, University of Dicle, 21280 Diyarbakir, Turkey


Plant tissue culture techniques tools in plant micropropagation

THANK YOU!


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