Micropropagation Lab Report

MICROPROPAGATION: PRINCIPLE
The practice of propagation of a plant under ‘aseptic conditions’, for the production of its clones, is known as ‘micropropagation’. It is a technique which comes under plant tissue culture. This technique allows large number of plants to be produced from the explants in less time. Through micropropagation, various ‘difficult-to-root’ species can be effectively reproduced clonally; but the cost is high which has resulted in its prevention for a wider application in market [32]. The following stages are involved during this process:
• Stage 0 – Preparative/ Pre-initiation stage: Explant selection and sterilization takes place.
• Stage I - Initiation (of aseptic culture) and Multiplication stage: Explant is placed

According to plant biology, the cells which cover a plant wound are callus cells. The induction of callus formation can be done by placing the plant tissues (after their surface sterilization) on the media in-vitro. . Many types of media are available but the ones which are most used are Murashige and Skoog (MS) medium [33], woody plant medium [34] and White's medium [35]. Various plant growth regulators are used for the initiation of callus, such as, equal amounts of cytokinin and auxin result in the production of this unorganized mass of cells. There are two broad categories of callus-cultures: friable or compact. Callus is very useful in micropropagation where it can be efficiently used to grow ‘genetically identical copies’ of plants with desirable

1 ml of NaOH (1N) was added to it. The powder was uniformly dissolved by shaking the tube gently. After this, the final volume was made up to 10 ml by addition of double distilled water. All this was done inside the Laminar Air Flow unit. If necessary, the tube was vortexed to ensure complete dissolution. Lastly, the tube was stored in the refrigerator (4 degree Celsius).
• The same procedure discussed above was followed for the preparation of 1mg/ml stock solutions of TDZ and 2, 4- D.

PREPARATION OF STOCK SOLUTIONS OF MURASHIGE-SKOOG MEDIA:
The nutrient formulation which supports the plant-growth (either liquid or solid: gel- like) is known as plant-growth media. The growth of the plant is governed by this media in-vitro. The same nutrients, as that of the plant grown in natural environment, are required by the plant in culture. But, some specific components are required for the promotion of optimum growth of the plant-culture in labs. The principle contents of the media include: carbon source, inorganic nutrients, organic supplements, growth regulators and a gelling agent. Murashige and Skoog medium (MS media) is the most commonly used ‘plant growth’ media for the purpose of plant-cell culture. It was invented in 1962, by T. Murashige and F. K. Skoog [37]. There are 4 stock solutions of MS media:

STOCK A (macronutrients)- 100X FOR 1L
KNO3