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Teknik & Aplikasi KuljarDalam Produksi & Kualitas Tanaman
Teknik & Aplikasi KuljarDalam Produksi & Kualitas Tanaman
“… the art and science of multiplying plants in vitro.”
“… the art and science of multiplying plants in vitro.”Dr. Sukendah, MSc
Pembentukan Benih Sintetik
Artificial seed coat
Somatic embryo
Artificial endosperm
Benih sintetik (Synthetic/Artificial Seed) adalah produksi benih dari embrio somatik yang dibungkus dalam mantel tiruan
- 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
Teknik Pembuatan Benih Sintetik
Bahan Benih:Embrio Somatik
Sodium alginateCalcium cloride
Seed Coat
Seed
Artificial Seed
3% Sodium alginate + MS free calcium salt solution
75 mM CaCl2.2H2O
Pembentukan seedcoat
Haploid Plant Production Embryo rescue of interspecific crosses
Creation of alloploids (e.g. triticale)Bulbosum method
Anther culture/Microspore cultureCulturing of Anthers or Pollen grains (microspores)Derive a mature plant from a single microspore
Ovule cultureCulturing 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 anthersCold or heat have both been effective
Culture mediaAdditives, 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 treatmentSpontaneous 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 fuseosmotic shock, electrical current, virus
Regenerate the hybrid fusion product
Contain genome from both organisms
Very, very difficult
Thank You