Transcript No Slide Title

Teknik & Aplikasi Kuljar
Dalam Produksi & Kualitas Tanaman
Dr. Sukendah, MSc
“… the art and
science of multiplying
plants in vitro.”
Pembentukan Benih Sintetik
Benih sintetik (Synthetic/Artificial Seed) adalah
produksi benih dari embrio somatik yang dibungkus
dalam mantel tiruan
Artificial
seed coat
Somatic
embryo
Artificial
endosperm
Teknik Pembuatan Benih Sintetik
- MS medium + phytohormone +
carbon
sources.
- Sodium alginate was added to the medium and
mixed well to make a homogenous solution.
- The explants were taken and dipped into the
alginate solution.
- Explants dipped in alginate solution were taken
up by a forcep and placed to the beaker of
CaCl2
- Each explant then became a hard ball encoded
by alginate.
Teknik Pembuatan Benih Sintetik
Bahan Benih:
Embrio Somatik
Seed
3% Sodium alginate +
MS free calcium salt
solution
Sodium alginate
Calcium cloride
Pembentukan
seedcoat
Seed Coat
75 mM CaCl2.2H2O
Artificial Seed
Haploid Plant Production

Embryo rescue of interspecific crosses
 Creation of alloploids (e.g. triticale)
 Bulbosum method

Anther culture/Microspore culture
 Culturing of Anthers or Pollen grains (microspores)
 Derive a mature plant from a single microspore

Ovule culture
 Culturing of unfertilized ovules (macrospores)
 Sometimes “trick” ovule into thinking it has been fertilized
Anther/Microspore Culture
Anther/Microspore Culture Factors

Genotype
 As with all tissue culture techniques

Growth of mother plant
 Usually requires optimum growing conditions

Correct stage of pollen development
 Need to be able to switch pollen development from
gametogenesis to embryogenesis

Pretreatment of anthers
 Cold or heat have both been effective

Culture media
 Additives, Agar vs. ‘Floating’
What do you do with the haploid?
Weak, sterile plant
 Usually want to double the chromosomes, creating a
dihaploid plant with normal growth & fertility
 Chromosomes can be doubled by

 Colchicine treatment
 Spontaneous doubling
○ Tends to occur in all haploids at varying levels
○ Many systems rely on it, using visual observation to detect
spontaneous dihaploids
○ Can be confirmed using flow cytometry
Protoplast
Created by degrading the cell wall using enzymes
Protoplast fusion
Protoplasts are made from two species
that you want to cross
 The membranes are made to fuse

 osmotic shock, electrical current, virus
Regenerate the hybrid fusion product
 Contain genome from both organisms
 Very, very difficult
