Micro-propagation Tissue Culture

Micro-propagation Ability to establish and maintain Plant organs & Plant tissues Done in aseptic culture Regenerate new plants

Micro-propagation (Tissue Culture) This is a technique of producing new plant from single cells, tissue, or small pieces of vegetative material (explants). The tissue obtained (mature or immature) is cultured in an artificial medium under sterile condition (invitro).

Stages First changes into a mass of undifferentiated cells called callus From the callus differentiation into shoot and roots or embryos may later occur. The technique can be used in both herbaceous and woody plants.

Basic Techniques Callus culture – mass of undifferentiated cells that can be induced or arise naturally as a result of wounding and growth substances Organ culture – the dedifferentiation of the parenchyma cells to produce centers of meristematic activity. Cell suspension culture – the disassociation of cells from each other on a liquid medium .

Facilities Required Preparation Area Transfer Area Growing Area Service Area

Methods involved Choosing a suitable explants Surface sterilization of the plant material Preparation of the stock solution –salts, growth regulators etc. Culturing Sub-culturing Removing plants from culture and place into suitable rooting medium

Developmental stages in micropropagation Stage I - Establishment Stage II – Multiplication Stage III – Root Formation Stage IV - Acclimatization

Flow Diagram

Advantages of Plant Tissue Culture Useful for rapid asexual multiplication of plants eg. Dracenas, carnations, orchids, ferns etc. Produce large quantities of a given variety/clone Produce plants which are slow and difficult to propagate Produce the ability to recover disease free material eg. From viruses.

Disadvantages Is costly Time consuming and difficult compared to other methods of plant propagation Require technical expertise or trained individual

Simple Micropropagation

Callus - dedifferentiate

Differentiation of callus tissue

Tissues on growth medium

Meristematic dome of a Plant

Important Terminologies Callus – an unorganized, proliferating mass of mostly undifferentiated cells Culture- a plant growing invitro Cell culture – culture of single cells Explant – part of a plant used to start a culture. Clones – plants produced asexually from a single plant.

Terminologies Growth regulator – organic compounds that influence growth and multiplication, such as cytokinins and auxins Invitro Exvitro Invivo Laminar air flow HEPA Growth room

Class Assignment List and explain the use of nine (9) equipment used in Tissue Culture. In order to develop a viable Tissue Culture unit which is successful. Four areas are required, name then and explain what happens in each area.

References Acquaah, G. (2009). Horticulture: principles and practices 4th ed. http://www.uky.edu/Ag/Horticulture/Geneve/teaching/plantpropwebpages/micropropagation/tctypesmicropropagation.pdf

THE END Courtesy:M.Gager, CASE