Tissue Culture Services

General

Our services start from. ………………….

• Searching a suitable land according to the expectation & assisting to acquire the land
• Levelling of the land
• Erection of fencing
• Digging of borewells
• Designing and Construction of Plant Tissue Culture laboratory.
• Designing and Construction of Poly house and shade house.
• Procuring of Laboratory equipments.
• Purchasing of Chemicals.
• We will identify suitable Mother plant for research and production purpose
• We will prepare a work plan for research and production purpose.

Technical

Ex-plant Sterilization:

• Collect the ex-plants from the source (Mother Plant) (hedge garden)
• Trim the ex-plants properly and taken in a sterile containers.
• Washed in running tap water 10 min until dirt gets dispense.
• Pour the ex-plants in a container with detergent solution upto 10 min (Tween – 20 or Teepol solution) i.e., till to reduce surface tension.
• Wash the ex-plants thoroughly and place it in fungicidal & antibiotic solutions upto one hour.
• Wash it and place in PVP solution (only for woody species) to absorb secondly metabolites.
• Enter into the sterile surface area (inoculation room)
• Wash the ex-plants in ascorbic & citric acid (0.1 g / lt) solution for 2 mins.
• Wash with sterile water.
• Dip the ex-plants in 70% alcohol for 30 seconds.
• Then wash the ex-plants in HgCl2 (0.1%) for 3 minutes.
• Wash the ex-plants with sterile water and tunic the ex-plants and leave for drying up to 10 mins.
• Inoculate the ex-plants in the sterilized medium.

MULTIPLICATION:

• Cut the newly sprouted shoots and placed on sterilized medium for multiplication (Sub culturing for multiplication).
• For every 4-6 weeks the plant material should be transferred to fresh medium.
• Before transferring onto fresh medium cut all the newly sprouted shoots into nodes and shoot tips
• Incubation period 4 – 6 weeks – stage / cycle.
• Subculture for multiplication for every 4-6 weeks to multiply the number i.e., up to 15-20 cycles.

ROOTING

• Rooting is of two types.
  • In vitro rooting
  • Ex vitro rooting
• In vitro rooting: cut the elongated shoots of 4 – 6 cm length and placed on rooting medium.
• Ex vitro rooting: place the shootlets in water and then in fungicide (0.1% Bavistin) solution for 15 – 20 mins.
• Dip the shootlets in rooting hormone.
• Plant the shootlets in root trainers containing vermiculate.
• Formation of roots after 4 weeks can be observed in both in vitro and ex vitro rooting.

HARDENING

• Pre hardening: place the bottles containing in vitro rooted shoots in mist chamber for 4 weeks.
• After 4 weeks take out all the in vitro rooted shoots and place the shoot lets in water and then in fungicide (0.1% Bavistin) solution for 15 – 20 mins.
• Transfer all the rooted plants into root trainers containing soil, coir peat, compost and vermicompost.
• Place these root trainers in mist chamber for 4 weeks.
• After 4 weeks transfer the root trainers from mist chamber to lath house.
• Secondary hardening: The plants are transferred into poly bags with in 25 days the poly bags contain 10 parts of red soil, 5 parts of compost and 2 parts of sand.
• These bags are kept in the poly house for maintain relative humidity and temperature. It takes 10 days for initial hardening stage.
• Then the poly bags are kept outside the poly house.
• This hardening treatment is given to the plants because, they get adjusted to the out side environment after transplantation, plants are watered once every day.

Plant protection

• The plants in the nursery are to be protected from all possible pests. Normally application of furadan 3G combined with good nursery hygiene would keep the pest problem of white grubs, termites and crickets under check. Defoliators and sap sucking insects are the usual pests of teak.
• For the control of defoliators and sap sucking pests spray Nuvacron @ 1 ml/litre of water (or) Metasystox @ 1 ml/litre of water (or) Hostathion @ 1 ml/litre of water (pr) Tafethion @ 1 ml/litre of water.

TRANSPLANTATION:

• Now the plants are ready for planting, before transplantation, 1 – Foot depth pits of 1x1 size pits are done they exposed to sun for killing the soil born pathogens.
• The dump in the pit,1 part of red soil and 1 part of manure and planting the plants in the center of the pit.
• Irrigation done on alternative days for 2 months and after that irrigation is done for 10 days from there onwards.