Clonal Forestry
TISSUE CULTURE TECHNIQUES FOR TREE SPECIES

Facilities required for mass multiplication : Any full fledged tissue culture laboratory should need the following infrastructural facilities.

  • General working laboratory
  • Media preparation room
  • Inoculation room
  • Culture room
  • Hardening chamber/shade house

a) General Working Laboratory : This laboratory is meant for general purpose working which should contain the following

  • Double distillation unit
  • Hot air oven
  • Autoclave
  • Storage cabinet
  • Sink for washing

b) Media Preparation Room : This room should contain

  • Electronic balance
  • Microwave oven (gas connection)
  • Autoclave
  • pH meter
  • Magnetic stirrer
  • Refrigerator/deep freezer
c) Inoculation Room
A small dust tree preferably airconditioned room equipped with an overhead UV lamp and a laminar flow chamber will serve as the inoculation area. A compound microscope with photographic accessories is an optional requirement. A small anti room provided with overhead airshower/air curtain before entry in to the inoculation and culture area should be of added advantage.
d) Culture Room
All types of tissue culture are to be incubated under well controlled temperature, humidity, air circulation, light quality and duration. The temperature should be maintained around 25 ± 2º C with air conditioners controlled by temperature controller. The culture room should be provided with racks with florescent lamps ranging from 1000-10,000 lux with facility to adjust the photoperiod. A lux meter to regularly monitor the intensity of the light is necessary. Shakers and dark chambers may also be required for some species. A generator to maintain uninterrupted power supply is optional requirement.

e) Hardening Chamber
A mist house with controlled fogging system will help in gradual weaning of in vitro raised plants. Then, plantlets are exposed to different light regimes under shade house conditions.

Problems in Micropagation of trees

Tissue culture of trees unlike other horticultural plants is beset with very special problems. Some of these include the physiological nature of the material (Juvenile and mature phases), general recalcitrant response of the explants vis a vis medium, inadequate rooting of the regenerated shoots and the associated problems of poor transfer ratio of established plants into soil. Most of the problems arising at tissue culture level can indeed be sorted out but commercial exploitation of techniques developed for tree tissue culture into a technology calls for concerted efforts of tissue culturists, foresters and tree breeders.

Reference: Surendran, C., Parthiban, K.T., Vanangamudi, K., and Balaji, S., 2000. Vegetative Propagation of Trees, Principles and Practices.1-154, TNAU press,

 

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