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Teknik & Aplikasi Kuljar

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Presentation on theme: "Teknik & Aplikasi Kuljar"— Presentation transcript:

1 Teknik & Aplikasi Kuljar
Dalam Produksi & Kualitas Tanaman “… the art and science of multiplying plants in vitro.” Dr. Sukendah, MSc

2 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

3 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.

4 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

5 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

6 Anther/Microspore Culture

7 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’

8 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

9 Created by degrading the cell wall using enzymes
Protoplast Created by degrading the cell wall using enzymes

10 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


12 Thank You

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