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Culture contamination
Vitrification
Acclimatization
Post culture behavior
Culture Contamination
Two sources:
Specific tests:
ELISA Viruses and bacteria
DNA probes All organisms
Rapid diagnostic kit Bacteria
Fatty acid profiling Bacteria
Vitrification
Translucency
Hyperhydration
Succulency
Glassines
lower RH
higher light levels
autotrophic growth
septic environment
In vitro rooting
Disadvantage:
Lack root hair
Died and collapsed after plantlet was removed from culture,
however new lateral and adventitious root formed during
acclimatization
The transition zone between root and shoot was abnormal
The vascular connection were poorly form
Restricted water uptake
Labor intensive and expensive
Ex-vitro rooting
Direct rooting during acclimatization
Cause:
When shoots are derived from dedifferentiated cell
Rapid proliferation of single cells or multi-cellular primordia through
organogenesis or embryogenesis
In vitro process or by added biochemical and stress agent
Temporary or heritable deformities
Cause
Variation that existed in the source plant
Chimera
Non chimeric chromosomal variation
Genetic changes
Mitotic abnormality
Somatic crossing over
polyploidy
Epigenetic or physiological effect
A non heritable change in phenotype that occurs in a substantial
percentage of the propagated population through an inducible
directed and reversible process
Chimera
Plant or plant part composed of genetically
different sort of cells as a result of mutation
or grafting
Plant with two or more distinct genotypes
Chimera
Cell origin
Mericlinal chimera
a section of one or two of the histogenic layers are
different
Sectorial chimera
all histogens in a sector are different
Periclinal chimera
one histogen is different from the others
Mimicked by variegated or mosaic forms
Due to en-even distribution of viruses in plant
tissue
Histogen
Cell layers in all higher plant tissue that trace back to distinct
layers in the apical meristem
3 layers in angiosperm
LI an outer epidermal layer
LII an internal tunica layer
LIII a cortical layer
LII layer produce gametic tissue and some surrounding
maternal tissue
The remaining maternal tissue is also formed by LIII and LI
Root derived from LII and LIII layers
GROWING POINT (APICAL MERISTEM)
Layer Gives rise to:
L-I Epidermis of all organs;
Monocot leaves - L-I contributes to the outermost region of
the leaf mesophyll giving rise to a strip along the leaf
margin.
Dicot leaves - L-I usually gives rise to only the colorless
epidermis, thus cannot be seen; sometimes L-I gives rise to
small islands of tissue along the margin
L-II Stem and roots:
Outer and inner cortex and some of vascular cylinder leaves:
mesophyll in outer region of leaf
L-III Stem and roots:
inner cortex, vascular cylinder and pith leaves: mesophyll in
central region of leaf
Non chimeric
chromosomal variation
Breakage in heterochromatic region
Somatic crossing over (mitotic exchange
between homologous chromosomes)
Gene amplification due to mutagenic
agent
Permanent genetic change
Somaclonal variation
Genetic change
Polyploidy Aneuploidy and breakage
Micronucleus formation Bi- or multi nucleate cells
Duplication Recombination
Inversion Amplification
Simple base pair change Organelle genome variation
Isozyme differences
Expression of cryptic transposable element
Change in chromosome structure
Definition
• Euploidy
An even increase in number of genomes (entire
chromosome sets)
• Aneuploidy
An increase in number of chromosomes within a
genome
Euploid
Euploid Symbol Somatic (2n)
monoploid x (ABC)
diploid 2x (ABC)(ABC)
triploid 3x (ABC)(ABC)(ABC)
autotetraploid 4x (ABC)(ABC)(ABC)(ABC)
(missing 2 different
double monosomic 2x-1-1 (AB)(AC)
chromosomes)
(2 additional different
double trisomic 2x+1+1 (ABC)(ABC)(A)(B)
chromosomes)
(2 additional chromosomes -
tetrasomic 2x+2 (ABC)(ABC)(A)(A)
same)
(missing a chromosome +
trisomic-monosomic 2x+1-1 (ABC)(AB)(A)
additional chromosome)
Plant variation from
dedifferentiated cell
Mitotic asynchrony caused by growth regulator
effect on DNA biochemistry (2,4,5-T; 2,4-D;
antibiotic; alkaloid; physical mutagen)
Disorientation or dysfunction of the mitotic
apparatus (spindle fiber)
Selection pressure due to the change in plant’s
environment
2,4-D
Juvenility:
The condition of a seedling plant that prevents
flowering or sexual gameto-genesis