TOPIC 9: PLANT CELL AND TISSUE CULTURE Flashcards
Plant cell culture
Plant tissue culture is a collection of techniques used to maintain or grow plant cells, tissues, or organs, under sterile conditions on a nutrient culture medium of
known composition.
Plant tissue culture is widely used to produce clones of a plant in a method known as micropropagation
Explant
Living tissue transferred from a plant to an artificial
medium for culture. It can be any portion of the shoot, leaves, roots, flower, or cells from a plant.
How does explant works?
Adult plant cells are totipotent, meaning they have the ability to give rise to a fully differentiated plant. Because of this, it is
possible to collect cells from a mature plant and use those cells to produce clones of that plant.
3 fundamental principals:
• The plant parts must be isolated from the rest of the body
• The plant parts (explant; isolated cells that is not affected) must be in a controlled, preferably defined, nutrient medium
• Aseptic conditions must be maintained
Types of plant cultures
• Callus culture
• Cell suspension culture
• Anther culture
• Ovule culture
• Embryo culture
• Protoplast culture
• Micropropogation
Callus Culture Techniques
When an excised and isolated piece of tissue is cultured on a nutrient medium, an unorganized mass of cell appears, is called callus. This callus is transferred on to different media to regenerate plants. This technique is called tissue culture.
Cell Suspension Culture
It is the culture of isolated cells or very small cell aggregates dispersed in liquid medium. The cell suspension is obtained by agitating pieces of callus in liquid medium on gyrating shaker.
Anther/Pollen Culture
Anther culture is a mean to produce plants with a gametic number of chromosomes by aseptic culture of anther. The technique give rise to haploid plants either directly or through
Ovule Culture
Ovule culture techniques is an important technique in modern plant breeding. it is much easier to culture whole ovule than to isolate a single embryo, especially in small seeded plants. E.g., tobacco
Embryo Culture
The embryos are isolated from young seeds and placed on a solid medium containing nutrients and vitamins. Embryos are cultured at 25°c,first in dark until seedlings are about 2 cm long and root formation has started, and than in light
Protoplast Culture
It is one of the most significant and recent developments in the field of plant tissue culture. the protoplast are usually isolated from cultured cell or leaf mesophyll cell by treating them with enzyme solutions. the isolated protoplast may be
used to regenerate the plants directly, or for the production of somatic hybrids through fusion.
Micropropagation
It is in vitro asexual propagation of crop plants. This
technique is advantages over the conventional practice of asexual propagation as only a small amount of plant is needed, species highly resistant to conventional bulk propagation can be propagated by this method and it is non season dependent.
Micro propagation is used for rapid multiplication of stocks, elimination of diseases, germ plasm preservation, and induction of mutation.
3 important aspects in vitro culture
1) Nutrient medium
2) Aseptic condition
3) Aeration of the tissues
Nutrient Medium
1) Depends upon the type of plant tissues or cell used for culture
2) Generally nutrient consists of
i) Inorganic salts
ii) A carbon source
iii) Vitamins
iv) Amino acids
Aseptic condition
1) Nutrient medium contains sugar which increases growth of microbes
2) These microbes compete with growing tissue and finally kill it
3) It is important to maintain aseptic condition
4) Sterilization is very important to stop the growth of microbes
