Teknik & Aplikasi KuljarDalam Produksi & Kualitas Tanaman

“… the art and science of multiplying plants in vitro.”Dr. Sukendah, MSc

Pembentukan Benih Sintetik

Artificial seed coatSomatic 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

ProtoplastCreated 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

Thank You