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Media preparation room, Stage 0: Selection & Preparation of explant,…
Media preparation room
Aseptic transfer room
Primary growth room
Store Growth room
Washing room
Stage 0: Selection & Preparation of explant
Stage 1: Sterilization and preparation of media
Stage 2: Multiplication
Stage 3: Rooting
Stage 4: Hardening/ Acclimatization
Deflasked and gradually decreasing the humidity
young plant with no cuticle will cause water loss
Environment factor
Light(photoperiod): 16day: 6 night
Temperature : 25+/-2
Sterility
Media
Frequent Subculturing
Avoid shortage of nutrient & build up of metabolic toxic
Sterilization
Dry heat
Moist heat
Chemical
Ray
Ultrafiltration
Sensitive media(GA, enzyme, antibiotic)[cellulose filter]
Plastic ware(UV ray)
Plant tissue(sodium hypochloriteðanol)
Concentration
Duration
Types of sterilant
Combination
Autoclave[121C &103kPa]
Metal appratus(except knife)& glasswear(not plastic)
MS media preparation
MS media
Macronutrient
Micronutrient
Vitamins
PGR
Auxin(2,4-D; NAA): stimulates root growth
Cytokinin(6-benzylaminopurine, kinetin): stimulate shoot growth
GA
etylene
Abscisic acid
thiamin, niacin, pyridoxine
FeMoMnCuIBZn
NPK CaMgS
White media
LS medium
Support
Agar, Gelrite
Organic additive
Coconut water
Sugar
Sucrose
Less differentiated, young explant
Genetics (diff genotype will responses diff with diff treatment)
Cell suspension culture
Callus culture
Embryo culture
Surface disinfaction
Trim
Soak with strilant
Rinse with water