Somaklonal variation

Zulfahmi
 Definition
Somaclonal variation is the variation seen in
plant that have been produced by plant tissue
culture
 Types of somaclonal variants
genetic changes
point mutations (e.g., Adh mutants in wheat)
cytoplasmic (maternal inheritance)
gene amplification (e.g., incr. gene copy no.)
activation of transposable element
 Types of somaclonal variants
genetic changes
cytogenetic (changes to genome structure)
aneuploidy gain or loss of 1 or more chromosomes
polyploidy gain or loss of an entire genome
translocation arms of chromosomes switched
inversion piece of chromosome inverted
 Types of somaclonal variants
epigenetic
change in phenotype that isn't stable during sexual
propagation
may or may not be stable during asexual propagation
usu. undesirable in a breeding program, not always
undesirable in propagation
habituation (most studied epigenetic change)
 Types of somaclonal variants
epigenetic
habituation (most studied epigenetic change)
defn loss of exogenous requirement for a growth factor (
usu. a PGR); e.g., auxin, cytokinin habituation
detection callus may lose requirement for a PGR in the
process of several transfers to fresh medium
 Types of somaclonal variants
epigenetic
characteristics
often occur gradually
are regularly reversible (esp. in regenerated plants)
are not seed-transmitted
 Possible causes
the dedifferentiation redifferentiation process
axillary shoot proliferation vs. organogenesis &
embryogenesis
hypothesis of D'Amato
somaclonal variants are rare in micropropagated plants (when
multiplication is by axillary branching of shoot tips/buds)
more common during shoot organogenesis & somatic
embryogenesis (esp. w/a callus phase)
 Possible causes
the "culture environment"
the hypothesis that tissue culture is inherently
stressful to cultured plant cells
environmental stress is known to cause:
DNA methylation the methylation of cytosine is known to
cause gene inactivation; this may occur during the
redifferentiation process
gene amplification can result in incr. gene expression
 Possible causes
the "culture environment"
environmental stress is known to cause:
transpositional changes
inadequate control of the cell cycle (errors in microtubule
synthesis, spindle formation)
importance of PGRs
scant evidence of direct mutagenic action
more evidence for transient modifications of phenotype
(e.g., dwarfing)

Dalam kultur jaringan Mutasi yang terjadi adalah
mutasi somatik

Sangat dipengaruhi oleh kondisi sel itu sendiri

Sel yang mengalami mutasi Melakukan pembelahan
Membentuk sel yang berbeda dengan sel asalnya
Tanaman yang berbeda

Kultur Jaringan merupakan teknik perbanyakan
vegetatif, hal ini membuktikan teknik vegetatif dapat
menghasilkan tanaman yang tidak sama dengan tetuanya.

Kultur Jaringan Memiliki potensi yang sangat besar
dalam meningkatkan keragaman.
Li & Gray, (2005) Faktor-Faktor Yang mempengaruhi
Keragaman Somaklonal

Faktor Fisiologis

Perlakuan Auksin (2,4-D, 2,4,5-T) pada tanaman kelapa sawit
menghasilkan buah yang bersayap

Faktor Genetik
 Keragaman genetik selama proses regenerasi kultur jaringan
merupakan perubahan pada tingkat kromosom
 Fenotipe Seolah-olah sama tetapi secara genetik berbeda
 Penempatan Heterokromatin yang terlambat menyebabkan
terjadinya keragaman somaklon pada tan jagung

Faktor Biokimia
Faktor Biokimia merupakan faktor yang paling banyak terjadi
(Jayasankar, 2005)
Metabolisme carbon menyebabkan fotosintesa lintasan
karoten, biosintesa pati, Metabolisme nitrogen
menjadi rendah
 Induksi Keragaman somaklonal secara eksogen.
1). Pengaturan Penanaman eksplan
Pengaturan ZPT
Lamanya Pengkulturan
Organogenesis langsung
Induksi Kalus
Kultur Protoplas
2). Mutagen Penginduksi Keragaman Somaklonal
Kimia: (Kolkisin, EMS(Etil Metan Solfonat),DEMS(Dietil Metan
Sulfonat)DES(Dietil slfonat)EL(Etilen Amin), ENH(Etit Nitroso Urea), MNH
(Metil Nitroso Urea)

Fisik : Sinar X, Sinar Gamma,

 Somaclonal variation leads to the creation of
additional genetic variability. Characteristics
for which somaclonal mutants can be
enriched during in vitro culture includes
resistance to disease pathotoxins, herbicides
and tolerance to environmental or chemical
stress, as well as for increased production of
secondary metabolites.
 Negative consequences (of somaclonal var.)
introduction of deleterious genes
dominant alleles albinos, pollen sterility
recessive alleles off-types that show up in progeny of
seed-propagated cultivars
carry-over effect from tissue culture
common with asexually propagated plants
symptom excessive and multiple branching of
regenerated plants (lack of establishment of apical
dominance)
 Applications to crop improvement
overcoming barriers to genetic exchange
embryo culture of interspecific hybrids (Lolium,
Hordeum)
selection for stable mitosis, fertility in a hybrid; e.g.,
intergeneric hybrids of barley x canada wild rye
seed cultivar improvement (e.g., Fusarium
resistance in celery, white-flowered Torenia,
tomato cultivars with higher soluble solids, better
"flesh" color)
 Identifikasi keragaman somaklonal

1). Konvensional (Karakterisasi Morfologi)

2). Analisis Molokuler
RAPD (Ramdom Amplified Polymorphic DNA)
RFLF (Restriction Fragment Length
Polymorphism)
AFLF ( Amplified Fragment length
Polymorphism)
3). Analisis Isoenzim