plant tissue culture n.
Download
Skip this Video
Loading SlideShow in 5 Seconds..
Plant Tissue Culture PowerPoint Presentation
Download Presentation
Plant Tissue Culture

Loading in 2 Seconds...

play fullscreen
1 / 62

Plant Tissue Culture - PowerPoint PPT Presentation


  • 152 Views
  • Uploaded on

Plant Tissue Culture. Original by Linda Rist Modified by Georgia Agricultural Education Curriculum Office July, 2002. T.C. Refers to technique of growing plant cells, tissues, organs, seeds or other plant parts in a sterile environment on a nutrient medium. History.

loader
I am the owner, or an agent authorized to act on behalf of the owner, of the copyrighted work described.
capcha
Download Presentation

PowerPoint Slideshow about 'Plant Tissue Culture' - arlen


An Image/Link below is provided (as is) to download presentation

Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author.While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server.


- - - - - - - - - - - - - - - - - - - - - - - - - - E N D - - - - - - - - - - - - - - - - - - - - - - - - - -
Presentation Transcript
plant tissue culture

Plant Tissue Culture

Original by Linda Rist

Modified by Georgia Agricultural Education Curriculum Office

July, 2002

slide2
T.C.
  • Refers to technique of growing plant cells, tissues, organs, seeds
  • or other plant parts in a sterile environment on a nutrient medium
history
History
  • In 1902 Haberlandt proposed that single plant cells could be cultured
haberlandt
Haberlandt
  • did not culture them himself
1930 s
1930’s
  • White worked on T.C.
  • discovery of plant growth regulators
1930 s1
1930’s
  • importance of vitamins was determined for shoot and root culturing

A,D,E,K,C, B complex

1930 s2
1930’s
  • Indole-Acetic Acid
  • IAA
  • discovered in 1937
slide8
IAA
  • 2,4-D
  • Dicamba
  • NAA
  • IBA
  • all synthetic hormones
1957 58
1957-58
  • Miller and Skoog
  • University of Wisconsin - Madison
  • discovered Kinetin
kinetin
Kinetin
  • a cytokinin
  • plays active role in organogenesis
slide11
1958
  • Steward developed somatic embryo from carrot cells
1958 60
1958-60
  • Morel cultured orchids and dahlias
  • freed them from a viral disease
slide13
1962
  • Murashige and Skoog
  • published recipe for MS Medium
60 s 70 s
60’s & 70’s
  • Murashige cloned plants in vitro
  • promoted development of commercial plant T.C. labs
slide15
1966
  • raised haploid plants from pollen grains
slide16
1972
  • used protoplast fusion to hybridize 2 species of tobacco into one plant
  • contained 4N
slide17
4N
  • all chromosomes of both plants
70 s 80 s
70’s &80’s
  • develop techniques to introduce foreign DNA into plant cells
  • beginning of genetic engineering
t c media
T.C. Media
  • functions
  • provide H2O
  • provide mineral nutritional needs
t c media1
T.C. Media
  • provide growth regulators
  • Provide vitamins
  • provide organic compounds
t c media2
T.C. Media
  • provide access to atmosphere for gas exchange
  • serve as a dumping ground for plant metabolites
t c media3
T.C. Media
  • H2O is usually distilled
  • minerals must provide 17 essential elements
  • energy source and carbon skeletons - sucrose is preferred
vitamins
Vitamins
  • thiamine
  • pyridoxin
  • nicotinic acid
  • biotin
vitamins1
Vitamins
  • citric acid
  • ascorbic acid
  • inositol
growth regulators
Growth Regulators
  • auxins and cytokinins
  • gibberellic acid
  • abscissic acid
ph of media
pH of media
  • usually 5.0-5.7
media
Media
  • must be sterile
  • autoclave at 250 F at 15 psi for 15 minutes
t c stages
T.C. Stages
  • Explanting- Stage I
  • get plant material in sterile culture so it survives
  • provide with nutritional and light needs for growth
stage ii
Stage II
  • rapid multiplication
  • stabilized culture
  • goal for a commercial lab
  • difficult and time consuming to maintain
stage ii1
Stage II
  • occurs in different pathways in different plants
rooting stage iii
Rooting - Stage III
  • may occur in Stage II
  • usually induced by changes in hormonal environment
  • lower cytokinin concentration and increase auxin
rooting
Rooting
  • may skip stage III and root in a greenhouse
stage iv
Stage IV
  • transplantation and aftercare
  • usually done in greenhouse
  • keep RH high (relative humidity)
stage iv1
Stage IV
  • gradually increase light intensity and lower RH after rooting occurs
  • allows plants to harden and helps plants form cuticle
cuticle
Cuticle
  • waxy substance promotes development of stomates
  • plants in T.C. don’t have cuticle
explant
Explant
  • portion of plant removed and used for T.C.
  • Important features
  • size
  • source - some tissues are better than others
explant1
Explant
  • species dependent
  • physiological age - young portions of plant are most successful
explant2
Explant
  • degree of contamination
  • external infestation - soak plant in sodium hypochlorite solution
explant3
Explant
  • internal infection - isolate cell that is not infected
  • roots - especially difficult because of soil contact
explant4
Explant
  • herbaceous plants
  • soft stem
  • easier to culture than woody plants
patterns of multiplication
Patterns of multiplication
  • stage II - light 100-300 foot candles
  • callus - shoots - roots
  • stage III - rooting - light intensity 1000-3000 foot candles
genetic transformation
Genetic transformation
  • permanent incorporation of new or foreign DNA into genome of cell
transformation methods
Transformation methods
  • protoplast fusion
  • cell wall is removed by enzymes from cell
protoplasts
Protoplasts
  • naked plant cells
  • from 2 different plants can be mixed together and forced to fuse
protoplast fusion
Protoplast fusion
  • results in heterokaryon
  • cell containing two or more nuclei from different cells
  • homokaryon - from same cell
protoplast fusion1
Protoplast fusion
  • allowed to regenerate cell wall and then grow into callus
  • callus turns to shoots
shotgun approach
Shotgun approach
  • DNA coated micro bullets of gold or tungston
  • shot into growing cells
  • DuPont holds the patent
shotgun approach1
Shotgun approach
  • injures cells
  • random success rate
slide49
PEG
  • Polyethylene glycol
  • pores open similar to electroporation
ti plasmids
Ti Plasmids
  • Tumor inducing
  • Agrobacterium temefasciens
  • infect cells with agrobacterium which contains desired DNA
ti plasmids1
Ti Plasmids
  • monocots resist agrobacterium infection
  • researchers are working to overcome this
luciferase
Luciferase
  • an enzyme
  • put into tobacco using Ti plasmid
luciferase1
Luciferase
  • when transformed tobacco plants are watered with solution containing Luciferin
  • they break it down and emit light
luciferase2
Luciferase
  • glowing in the dark
  • like a fire fly
screening techniques
Screening techniques
  • used to identify if culture has taken on desired new trait
examples
Examples
  • sensitivity to antibiotics
  • color
  • sensitivity to excess deficiencies of substances in growth media
conventional
Conventional
  • plant breeding
  • egg cell gives half the chromosomes and almost all of the cytoplasm
  • male only gives its chromosomes
slide58
Cont…….
  • This condition is called maternal cytoplasmic inheritance
microinjection
Microinjection
  • single cells from culture are held stationary with gentle suction
  • injected with a tiny syringe loaded with DNA
microinjection1
Microinjection
  • done under electron microscope
electroporation
Electroporation
  • desired DNA in solution outside cell
  • high energy pulses - 50,000 volts
  • for a millisecond
electroporation1
Electroporation
  • cause tiny pores to open
  • allows DNA to enter the cell