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Teknik & Aplikasi Kuljar Dalam Produksi & Kualitas Tanaman. “… the art and science of multiplying plants in vitro .”. Dr. Sukendah , MSc. Pembentukan Benih Sintetik.
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Teknik & AplikasiKuljar DalamProduksi & KualitasTanaman “… the art and science of multiplying plants in vitro.” Dr. Sukendah, MSc
PembentukanBenihSintetik Benihsintetik (Synthetic/Artificial Seed) adalahproduksibenihdariembriosomatik yang dibungkusdalam mantel tiruan Artificial seed coat Somatic embryo Artificial endosperm
TeknikPembuatanBenihSintetik • 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.
TeknikPembuatanBenihSintetik BahanBenih: EmbrioSomatik Seed 3% Sodium alginate + MS free calcium salt solution Sodium alginate Calcium cloride Pembentukanseedcoat Seed Coat 75 mMCaCl2.2H2O 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 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