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Tissue Culture

Course CodeBHT306
Fee CodeS3
Duration (approx)100 hours
QualificationStatement of Attainment


Learn to Grow Plants with Tissue Culture

  • Grow Ferns, Orchids, Cut Flowers or Other Plants
  • Produce large quantities of new varieties fast and relatively cheap
  • Develop a High Tech small business at home
  • Discover what is possible with minimal investment using only a backyard garden

Tissue culture involves growing plants from very small sections (sometimes microscopic) in a laboratory. It is a propagation method which is being increasingly used. Tissue culture is not appropriate for many plants, but for others such as orchids, some indoor plants and in particular, many new plant varieties, it is a very popular propagation method.

Lesson Structure

There are 9 lessons in this course:

  1. Introduction including a review of basic plant nutrition.
    • Stages in tissue cultured plant development
    • Introduction to Plant Growth Science, biochemical processes and cell biology
    • Transpiration, Photosynthesis and Respiration
    • Plant Parts -Stems, Leaves, Roots, Buds,Flowers and fruits
    • What happens as Tissue Matures
    • Types of Plant Tissue
    • Methods of Shoot Induction and Proliferation
    • Advantitious Roots
    • Terminology
  2. Plant Nutrients
    • Major Elements
    • Minor (Trace) Elements
    • Total Salts
    • How Plants Grow
    • Factors Affecting Nutrient Uptake
    • Nutrient Solution Preparation
    • Hydroponic Nutrients
    • Chelates
    • Growing Media for Tissue Culture
    • Water in Tissue Culture
    • Chemical Analysis
  3. The Laboratory
    • The Tissue Culture Laboratory
    • Preparation Area
    • Transfer Chamber
    • Culture Growing Area
    • Siting a New Lab
    • Equipment Requirements for a Lab
    • Chemicals
  4. Micropropagation Techniques
    • Stock Plants -selection, planting, management
    • Uses for Tissue Culture
    • Problems with Tissue Culture
    • Procedures
    • Explants
    • Sterilisation
    • Nutrient Media
    • Shoot Induction and Proliferation
    • Rooting and Planting Out
    • Stages in Plant Development
    • Treating Plant Tissue with Sterilants
  5. Plant Hormones
    • Chemical Growth Modification
    • Principles of Using Plant Hormones
    • Auxins, Cytokinins, Gibberellins, Abscisic acid and Ethylene.
    • Other Chemical Treatments
  6. The Tissue Culture Environment
    • Media Types -Filter Bridge, Agar, Liquid
    • Nutrient Media Composition
    • Cleanlines
    • Light and Temperature
    • Hormones
    • Artificial Light
    • Water Quality
    • Water Treatgments
    • Carbon Dioxide Effects
    • Greenhouses
    • Diagnosis of Plant Disorders
  7. Commercial Applications
    • Understanding Genetics and Plant Breeding
    • Biotechnology
    • Cell Fusions
    • Overcoming Pollination Incompatibility
    • Pollination Biology
  8. Taking Plants out of Culture
    • Hardening off Plants
    • Growing Rooms or Chambers
    • Rockwool Applications with Micropropagation
  9. Culture of Selected Species
    • Begonia
    • Cattleya
    • Cymbidium
    • Review of a range of other plants

Each lesson culminates in an assignment which is submitted to the school, marked by the school's tutors and returned to you with any relevant suggestions, comments, and if necessary, extra reading.

Aims

  • Explain the nature of plant growth processes, in the tissue culture environment.
  • Determine growing media to use for tissue culture.
  • Specify appropriate micropropagation procedures for different purposes.
  • Explain the management of environmental control equipment used in tissue culture.
  • Design a layout for a commercial tissue culture facility.
  • Determine appropriate commercial applications for tissue culture.

What You Will Do

  • Describe botanical terms which may be relevant to tissue culture.
  • Explain different physiological processes which are relevant to tissue culture, including:
    • Photosynthesis
    • Transpiration
    • Respiration.
  • Differentiate between different types of plant tissue, including:
    • Collenchyma
    • Sclerenchyma
    • Parenchyma
    • Xylem
    • Phloem
    • Meristem.
  • Describe the stages of plant growth during tissue culture of a specified plant.
  • Explain the roles of the major and minor nutrients in tissue culture.
  • Explain how five different specified plant hormones can be used in tissue culturing plants.
  • Explain the functions of different types of components of media, including:
    • Nutrients
    • Carbohydrates
    • Vitamins
    • Growth regulators
    • Amino acids
    • Antibiotics.
  • Differentiate between appropriate applications for both liquid and solid media.
  • Compare two different specified formulae for tissue culturing, formulated for two different plant genera.
  • Explain fifteen different terms relevant to micropropagation procedures, including:
    • abscission
    • aseptic
    • autoclave
    • axenic
    • bridge
    • in vitro
    • deionize
    • differentiate
    • flaming
    • hardening off
    • indexing
    • pipette
    • precipitate
    • transfer
    • vitrification.
  • Describe different methods of shoot proliferation used in tissue culture.
  • Explain a method of sterilisation for plant tissue in an operation observed by you.
  • Distinguish between tissue culture operations which use different plant parts, including:
    • Meristem
    • Shoot tip
    • Organ
    • Cell.
  • Describe the steps in producing a plant by tissue culture, observed by you in a commercial facility.
  • Explain how to remove a specified plant from tissue culture, into open culture.
  • Compile a resource file of twenty different suppliers of environmental control equipment.
  • Determine guidelines for establishing an appropriate, controlled environment, for growing a tissue culture.
  • Describe two different greenhouse management methods for acclimatising tissue cultured plants.
  • Explain how knowledge of short-day, long-day and day-neutral plants is relevant to tissue culture.
  • Explain methods of ensuring water used in tissue culture is pure and sterile.
  • Determine the equipment needed to set up a tissue culture laboratory.
  • Describe the functions of the equipment listed.
  • Develop on-going maintenance guidelines for a tissue culture facility which has the range of equipment listed.
  • Determine consumable materials required for the day-to-day operation of a specified tissue culture facility.
  • Determine the minimum skills needed to set up a tissue culture laboratory.
  • Write a job specification for a tissue culture technician, which identifies skills needed in that job.
  • Draw a floor plan to scale, for a workable tissue culture laboratory, designed for a specified purpose.
  • Describe commercial micropropagation methods for three different plant genera.
  • Distinguish between the unique requirements for successful micropropagation of six different specified genera.
  • Analyse, from research, the use of tissue culture for plant breeding.
  • Determine criteria for assessing the commercial viability of using tissue culture for propagating a given plant.
  • Determine the number of plants of a specified plant variety which would need to be cultured, in order to make tissue culturing of that plant commercially viable.
  • Assess the commercial viability of a specified tissue culture enterprise.

What Investment or Facilities are Needed for Tissue Culture?

When someone starts a plant tissue culture business; they do not need  as much of an financial investment as with most other types of plant nurseries; largely because they do not need as much space (land).

There are nevertheless, certain things that you do need to have; somewhere sterile to propagate the plants, and a very clean environment in which to grow them on.

Commercially viable tissue culture businesses have been started in a space as small as a spare (converted) bedroom or a small backyard greenhouse.

Tissue Culture laboratories have operated successfully, employing a couple of full time staff in a business that uses as little as 500 square metres (dependant upon what is grown, how it is grown and at what stage it is marketed).

Once you have done this course, and better understand the process of tissue culture, you will be in a far better position to decide how to set up, what to grow, and how to grow plants with tissue culture techniques. You do not need to set up these facilities in order to complete this course. 

 

Buying or Constructing Growing Areas is often the Main Intitial Cost

A room where conditions can be controlled is the ideal facility for hardening off.

Temperature, humidity and light etc. can be gradually changed, allowing the plant to adapt through a series of small changes - as the environment evolves from a growing cabinet environment to an environment the same as the outdoors.

1. A simple growing room can be built in part of a greenhouse.

Temperature and light can easily be controlled by conventional heating and cooling equipment, and if necessary, artificial lights and shadecloth. Humidity can be controlled by partial or full coverage of containers with plastic tents or bell jars. Fungicides (such as Benlate) should be used at recommended intervals to control damping off & other fungal problems. (Damping off is one of the major dangers at this stage). An 11 inch X 21 inch tray covered with 0.4 ml plastic is a good unit to work with. Humidity is gradually lowered by first lifting one corner of the plastic cover and progressively removing more and more of the cover, as the plants harden.

2. Another method involves using a growing room as above, but instead of a plastic cover or tent over the plants, to use an automatic misting system to maintain humidity.

3. A third method of maintaining humidity is to sit containers of cultured plants in a bath (or tray) of water. The whole growing area might then be covered partially with a plastic tent, or a tent with openings at the ends.

4. Commercial Growth Cabinets of many types are available.

5. There is a trend towards rooting plantlets out of tissue culture in a growth cabinet or growing chamber. This has several advantages:

  • It is less expensive in terms of labour.
  • Stronger root systems tend to develop.
  • The use of the transfer room is reduced.
  • More space is made available in the growing room (A 50% peat/50% perlite mix has been successfully used for rooting begonias this way).

6. A cold Frame may in some cases provide an adequate transition stage from culture. ‑bottom heat may be needed to keep the roots developing.

Following a stage in a greenhouse, growth cabinet or some other structure as outlined above, tissue cultured plants would normally be then eased into the outside world....spending time in either (or both) a standard greenhouse and/or a shade house, before being placed into the open.