Escolar Documentos
Profissional Documentos
Cultura Documentos
Introduction
Regeneration of tissues cultures
Germplasm preservation In Vitro Approaches
Micropropagation techniques in horticulture and crop
improvement
Somatic hybridization
Key parameters for manipulation of plant tissue
cultures
Sterile techniques
Genetic manipulation of crop plants
Advantages of tissue culture over intact plants
Zygotic embryo
Seedling
Somatic embryos
Regenerant plants
Pathways for plant regeneration
Dedifferetiation Induction Realization
Aseptic Abundant
explant cell division
(Organs, tissues)
Callus + Determined Organgenesis
competent cells Cells or embryogenesis
Indirect
Protoplast regeneration
Abundant
No regeneration
cell division
Callus
From Su 2002
CSS451
Plant regeneration
Somatic embryogenesis--New plants are
formed from somatic embryos. Somatic embryos
are formed in plant tissue culture from plant cells
that are not normally involved in the development of
embryos, i.e. ordinary plant tissue.
CSS451
Organogenesis
Somatic Embryogenesis
a Embryogenic calluses
b Formation of somatic embryos
c Mature somatic embryos
d Plantlets
Proliferation
Proliferation
Rate
9.6 9.7 5.5
27.3 18.0 17.2
Regeneration
Germplasm Preservation In Vitro Approaches
Cold storage
storage at non-freezing temps, from 1-9 C dep. on spp.
storage of shoot cultures (stage I or II)
works well for strawberries, potatoes, grapes, and etc.
transferred (to fresh medium) every 6 mo. or on a yearly basis
1. Nutrient Media
2. Culture Explants
Provide water
Provide mineral nutrition
Provide vitamins
Provide growth regulators
Access to atmosphere for gas exchange
Removal of plant metabolite waste
Formulations of culture medium
Commercially,
Many available
Differ in salt concentrations
Differ in presence or absence of salts
Micronutrients
Macroelements
Iron (Fe) 1 m M
Manganese (Mn) 5-30 m M
Potassium (K) 20 -30 mM
Zinc (Zn)
Phosphorous (P) 1-3 mM
Boron (B)
Calcium (Ca) 1-3 mM
Copper (Cu) 0.1 m M
Magnesium (Mg) 1-3 mM
Molybdenum (Mo) 1 m M Plant growth regulators
Sulfur (S) 1-3 mM
Cobalt (Co) 0.1 m M
Element Function
Surface-sterilizing
Medium, tools, vessels plant materials
Alcohol
Autoclaving Bleach
Ethylene oxide gas Calcium hypochlorite
UV radiation Mercuric chloride
Dry heat Hydrogen peroxide
Microwave Rinsing
Autoclaving
ETOH /
2,4-D Autoclave 0.01-5.0
1N NaOH
Auxins 0.002- 2-4 times less active than
Picloram Autoclave Water
20.0 2, 4-D
NAA Autoclave 1N NaOH 0.1-10.0
Objectives:
For agar medium, cool down till to 50-60C; add antibiotics if required , mix well
and pour on the sterile plates.
Experiment 1: Plant tissue culture