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Plant Tissue Culture

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Plant Tissue Culture Maheswari, 1960 first anther culture Nitsch, 1974 microspore culture Cocking, 1960 Protoplast culture ... – PowerPoint PPT presentation

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Title: 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 (in vitro).
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
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25
Embryogenesis
26
Somatic embiryogenesis in cotton
27
History of plant tissue culture
  • TOTIPOTENSI 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
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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 in UNS
  • Shoot culture
  • Callus culture
  • Micro tuber
  • Somatic variability
  • Root culture
  • Plant improvement
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