TISSUE CULTURE

Learn to Grow Plants using Tissue Culture 

Lesson Structure

There are 8 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
   • Adventitious 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 Treatments
   • Carbon Dioxide Effects
   • Greenhouses
   • Diagnosis of Plant Disorders
7. Commercial Applications
   • Understanding Genetics and Plant Breeding
   • Biotechnology
   • Cell Fusions
   • Overcoming Pollination Incompatibility
   • Pollination Biology
   • Taking Plants out of Culture
   • Hardening off Plants
   • Growing Rooms or Chambers
   • Rockwool Applications with Micropropagation
8. Culture of Selected Species
   • Begonia
   • Cattleya
   • Cymbidium
   • Review of a range of other plants

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 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 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 different plant genera.
• Distinguish between the unique requirements for successful micropropagation of different 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.

HOW YOU CAN LEARN TO GROW ANYTHING WITH TISSUE CULTURE

A good knowledge of both tissue culture procedures, coupled with an appreciation of a particular plant's normal cultural requirements, should enable you to make a logical and calculated determination for an appropriate procedure and requirements to be applied in tissue culturing that plant.

Learn What Different Chemicals Can Do

A big part of tissue culture is learning what different chemicals can do to affect plant growth; then understanding how different plant species develop. Armed with that knowledge; the experienced micropropagator can set about determining what chemicals to use, when, where and with what plant species, to affect the way that plant develops in the tissue culture situation.
Consider just one type of chemical.

Gibberellins

At high concentrations, gibberellins inhibit root formation. If natural levels of gibberellin which occur in a plant are lowered, then root development should be improved. Chemicals such as "Alar" and "Arest" are commercial preparations which work by interfering with the affect of gibberellin in a plant, hence reducing the inhibition to certain types of growth.

Gibberellins are present in varying amounts in all parts of the plant, but the highest concentrations are found in immature seeds. The best known of the group is GA3, gibberellic acid.

Gibberellins stimulate cell division and cell elongation, and affect both leaves and stems. They have the following physiological effects:

• Stem elongation. Stems become long and thin, and leaves become pale when gibberellins are applied.
  
  
• Juvenility. Gibberellins promote juvenility when applied to the adult branches in plants that have distinct juvenile and adult phases.
  
  
• Flowering. Some rosette-forming plants produce elongated flowering stems after exposure to cold or long days (or both). When gibberellins are applied to such plants, they flower without the requirement for vernalisation or photoperiod. Gibberellin is thus used for early seed production of biennial plants such as cabbage.
  
  
• Pollen and fruit development. Gibberellins stimulate the pollen germination and the growth of pollen tubes in some plants. Similar to auxins, they can cause the development of parthenocarpic fruits, including apples, cucumbers and eggplants. Gibberellic acid is applied to commercial plantings of some grape varieties to create fuller, looser clusters of grapes.
  
  
• Seeds. Gibberellins overcome the need for dormancy-breaking cold or light requirements in certain plants, thus hastening seed germination.
  
  
• Root formation. At high concentrations, gibberellins inhibit root formation. If natural levels of gibberellins which occur in a plant are lowered, then root development should be improved. Chemicals such as "Alar" and "Arest" are commercial preparations that work by interfering with the effect of gibberellin in a plant, hence reducing the inhibition to certain types of growth.
  
  

There are over 130 different types of gibberellins. They are able to be classified both on the basis of either function or chemical structure.

In terms of chemical structure; there are two important groups of gibberellins:

• Nineteen carbon gibberellins  -These are biologically active forms. They are created from the 20 carbon types.
• Twenty carbon gibberellins.
  
  

The biological activity of a particular type of gibberellins is strongly affected by:

• number of hydroxyl groups linked to the carbon atoms
• location of these hydroxyl groups
  
  

Consider how much more complex ttissue culture can become when you are using all sorts of other chemicals, as well as gibberellins, and trying to manage the way they work together to control the development of thousands of plants in a laboratory environment

TISSUE CULTURE IS COMPLEX - IT DOES TAKE TIME TO LEARN, AND PRACTICE TO PERFECT

BUT ONCE YOU MASTER IT, YOU HAVE A SIGNIFICANT AND VALUABLE ADVANTAGE FOR MASS PRODUCTION OF NURSERY STOCK