Plant Tissue Culture | BiotechStudies

Plant Tissue Culture

What is Plant Tissue Culture?

•Plant Tissue culture is a technique of growing a part of plant on a nutrient medium under aseptic conditions. Part of plant used is called “explant”. It may be seed, cell, tissue, shoot apex, leaflet, root or any other part.
•Tissue culture involves both plant and animal cells
•The basic principle of plant tissue culture is Totipotency (the Ability of a plant cell to develop into a whole plant).

Why do Plant Tissue Culture?

•A single explant can be multiplied into several thousand plants in less than a year - this allows fast commercial propagation of new cultivars
•Taking an explant does not usually destroy the mother plant, so rare and endangered plants can be cloned safe.
•Disease free plant (new meristem tissue is usually virus free) can be cultivated to provide virus free plants
•Plant ‘tissue banks’ can be frozen, then regenerated through tissue culture.
•Mass propagation.
•Tissue culture clones are ‘true to type’ as compared with seedlings, which show greater variability.
•Conservation of Biodiversity.
•Easily Exportable.

Instuments used in PTC

•LAF It is necessary to provide sterile condition. It contain HEPA filters, UV lamp.
•Cultural vessel firstly autoclave and the used.
•Dissection Instruments and glassware sharp forceps and bent forceps.
•Autoclave.
•PH meter.
•Vortex, shaker
•Refrigerator, deep freeze.

What’s the Background?

•It is known as Father of plant tissue culture.
•Plant Tissue Culture had its origins at the beginning of the 20th century with the work of Gottleib Haberlandt.
•Firstly it cultured stamen hair cells ( Knops solution ).
•Enlargement of cells.
•Failed to induce cell division.
•(1838) schwann and schleiden gives totipotency theory.
•Hanning (1904) first attemp at embryo culture of selected crucifier.
•Robbins(1922) in vitro culture of isolated root tips.
•Skoog(1944) first in vitro culture of tobacco used to study adventitious shoot formation.
•Skoog and Tsui(1946) Formation of adventitous shoots and roots of tobacco determined by ratio of auxin: adenine
•Skoog and Miller (1957) regulation of organ formation(roots and shoots) by modifying cytokinin/auxin ratio. Higher ratio of auxin to cytokinin favoured root formation, the reverse shoot formation and that intermediate ratios promoted callus proliferation in tobacco callus.
•Murashinge and skoog (1962) The development of famous Murashige and skoog medium.

Laboratory and Practical Method

•Washing room
•Media preparation room
•Sterlization room
•Storage room
•Green house.

Case Study of Brahmi

• Mother Selection of Healthy plant
• Excision of explant (2cm)
• Treatment with tween20 (10 minutes)
• Rinsed with water (4-5 times)
• Treatment with 0.2% Bavistine and 0.2% sterptocycline (90 minutes)
• Rinsed with distilled water (4-5 times)
• Preparation of Media(Basal MS media +BAP +Sugar + Agar)
• Sterilization of Media (autoclaved)
• Storage of Media
• Inoculation in Laminar
• UV light on (10 minutes)
• Wiping with spirit
• Quick dip of Explant in 70% ethanol
• Rinsed with double distilled water (4-5 times)
• Sterilized Explant with 0.1HgCl2 (5 minutes)
• Rinsed with double distilled water (4-5 times)
• Inoculated in media
• Transferred to growth chamber

Application

•Disease free plant
•Formation of pharmaceuticals.
•Rejuvenation of plant material
•Anther culture
•Hybridization/ somatic Hybridization
•Bio transformation
•Embryo rescue
•Clonal propagation
•Somaclonal variation
•Germplasm conservation