Technique of In-Vitro Propagation Dr. Sukendah, MSc

Clonal in vitro propagation of plants from cells, tissues or organs cultured aseptically on defined media contained in culture vessels maintained under controlled conditions of light and temperature = MICROPROPAGATION Explant Cell, tissue or organ of a plant that is used to start in vitro cultures. Explant Cell, tissue or organ of a plant that is used to start in vitro cultures.

In vitro propagation Tissue culture propagation

Micropropagation Advantages Micropropagation Advantages From one to many propagules rapidlyFrom one to many propagules rapidly Multiplication in controlled lab conditionsMultiplication in controlled lab conditions Continuous propagation year roundContinuous propagation year round Potential for disease-free propagulesPotential for disease-free propagules

Conventional Propagation Advantages Equipment costs minimalEquipment costs minimal Experience can substitute for technical expertiseExperience can substitute for technical expertise Costs per unit are minimalCosts per unit are minimal Potential for disease-free propagulesPotential for disease-free propagules Specialized techniques for growth control (grafting onto dwarfing rootstocks)Specialized techniques for growth control (grafting onto dwarfing rootstocks)

Micropropagation Limitations Micropropagation Limitations Equipment/facility intensive operationEquipment/facility intensive operation Technical expertise in management positionsTechnical expertise in management positions Protocols not optimized for all speciesProtocols not optimized for all species Liners may not fit industry standardLiners may not fit industry standard Propagules may be too expensivePropagules may be too expensive

Micropropagation Applications Micropropagation Applications Rapid build up of stock of new, superior varietyRapid build up of stock of new, superior variety Elimination of endophytic diseaseElimination of endophytic disease Better suited to morphology/growth characteristics (few offshoots/sprouts; date palms, ferns, nandinas)Better suited to morphology/growth characteristics (few offshoots/sprouts; date palms, ferns, nandinas) Propagules have enhanced growth features (multibranched character; Ficus, Syngonium)Propagules have enhanced growth features (multibranched character; Ficus, Syngonium)

MACROPROPAGATION MICROPROPAGATION Small propagule Aseptic conditions Controlled environment Heterotrophic growth Rapid multiplication Greater initial costs Larger propagule Non-aseptic conditions Less environmental control Photoautotrophic growth Slower multiplication Nominal costs

 Perbaikkan formulasi media yang penting adalah pengembangan unsur makro yang universal, untuk mendukung pertumbuahan jaringan tumbuhan. Dalam media perlu ditambahkan ammonium dengan meningkatkan konsentrasi NO3- dan K +.  Media Knop  Media White  Media Knudson dan media Vacin and Went  Media Murashige & Skoog (media MS)  Media Gamborg B5 (media B5)  Media Schenk & Hildebrant (media SH)  Media WPM (Woody Plant Medium)