Plant Tissue Culture A rice plant growing in nutrient rich agar A.K.A Micropropagation In vitro Culture faperta.ugm.ac.id/newbie/download/pak_tar/Plantbiotechnology/TissuCulturebiotech2007.ppt

Plant Tissue Culture Learning Objectives Introduction to Tissue Culture Picture Dictation Exercise

Our Learning Objectives in relation to Tissue Culture Reasons for propagating plants by tissue culture are stated The culture process is described Describe some common tissue culture methods Including – meristem culture and embryo culture Describe how the plant part is selected Describe how the culture medium is made up Explain the timing of plant selection

Introduction to Tissue Culture Tissue Culture (also known as Micropropagation or In vitro culture) is: The growing of plant cells, tissues, organs, seeds or other plant parts in a sterile environment on a nutrient medium.

Introduction to Tissue Culture Plant tissue culture is a bit like the equivalent of ‘Dolly the sheep’ but using plants.

Introduction to Tissue Culture Cloning Animal Cells Cloning Plant Cells Both these processes use undifferentiated cells

Picture Dictation Exercise Take some meristematic cells from a plant. These cells are called an explant Place the explant on a sterile nutrient rich agar The explant grows into a ball of cells This ball of cells is called a callus The callus then develops roots, stem and leaves This is called a plantlet Transplant the plantlet into a traditional growing media Answer

Introduction to Tissue Culture Is …the technique of growing plant cells, tissues, organs, seeds or other plant parts in a sterile environment on a nutrient medium You might say … it is the plant equivalent of dolly the sheep Back to picture dictation Back to Contents

What are undifferentiated cells In the human body we have lots of different types of cells Blood cells Muscle cells Bones cells Brain cells An undifferentiated cell is a cell that does not yet have a specialised job Next

What are undifferentiated cells Undifferentiated cells in animals Stem cells Undifferentiated cells in plants ?

You tube demonstrations http://www.youtube.com/watch?v=kje0YczE0Do

The Culture Medium (Media) The environment surrounding the developing explants must provide the correct ‘necessities’ for plant growth. Neccessities for plant growth

The Culture Medium (Media) From this list of neccessaties – which ones are provided by the agar? Neccessities for plant growth

Hormones in the agar Two Hormones Affect Plant Differentiation: Auxin: Stimulates Root Development Cytokinin: Stimulates Shoot Development Generally, the ratio of these two hormones can determine plant development:  Auxin ↓Cytokinin = Root Development  Cytokinin ↓Auxin = Shoot Development Auxin = Cytokinin = Callus Development

What is Callus development A callus is a blob of tissue – (mostly undifferentiated cells) A callus is naturally developed on a plant as a result of a wound This callus can be left to develop or can be further divided

The culture medium contains a gel (agar) with the proper mixture of nutrients, sugars, vitamins and hormones (growth promotors), These causes the plant part to grow at very rapid rates to produce new plantlets.  It has been estimated that one chrysanthemum apex placed in tissue culture could produce up to 1,000,000 new plantlets in one year.

Tissue Culture Transfer Protocol   Dr. Dan Lineberger of Texas A&M University demonstrates the protocol to transfer African violets from tissue culture containers where they were grown into a small �forest� of cloned plants (called multiplication culture tubes) to tissue containers where the young clones will form new roots.� After the roots are formed, they can be removed and potted into containers.  This procedure must be done in the sterile environment of a transfer hood. Sterilize the surfaces of the transfer hood. Sterilize all tools that touch the plants by first dipping in alcohol them flaming. African violet clones in a shoot multiplication tube. Remove the cluster of plants in the culture. Insert the cluster of plants into the new culture container. Break up the cluster of plants and spread them out. Seal the container with paraffin film. The culture before transfer (left) and after transfer (right).

Embry Culture

Plant Tissue Culture and G.E