Tissue Culture Theory

Tissue culture is a method for multiplying plants by growing plant parts such as protoplasm, cells, groups of cells, tissues, or organs into complete plants under aseptic conditions. In 2014 Badak LNG began designing and building a plant tissue culture laboratory in collaboration with the plant tissue culture laboratory department of Biology, Faculty of Mathematics and Natural Sciences, Mulawarman University. The objective of the tissue culture laboratory program is to cultivate rare, endemic, or protected plants from Kalimantan in large quantity,  in order to increase biodiversity around the company. Until the end of September 2021 the number of living cultivated plants in the tissue culture incubation room reach 845 explants from 26 species (13 plants native to Kalimantan) while in the green house there are 711 plants from 16 plant species (8 plants native to Kalimantan).

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Tissue culture has a procedure consisting of: sterilization of tools and materials, preparation of media, initiation of culture, subculture/multiplication, and acclimatization.
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Step Description
1 Sterilization All tools must be Autoclaved first to eliminate contamination. Autoclave is a sterilizer with high pressure (17.5 psi) and high temperature (121 ͦC). This tool uses the principle of wet sterilization with high pressure hot water steam to keep the agar media in a good shape.
2 Media Preparation The planting media are the composition of basic media such as Murashige and Skoog (MS), Vacin and Went (VW), and Woody Plant Media (WPM) plus agar and sugar. In addition, sometimes researchers also put some growth regulator such as auxin and cytokinin with particular concentration to promote development of organs (leaves, stem, root).
3 Initiation Explant planting (initiation) is performed in the Laminar Air Flow Cabinet with aseptic conditions. Sterilized explants (ready for planting) are cut into pieces using scalpel and tweezers in a petri dish. The explant pieces are planted in a culture bottle that has filled the growing media. The culture bottle is closed with plastic and tightened with rubber.
4 Subculture Explants that have been successfully initiated (free from contamination of bacteria and fungi) can proceed to the subculture process. This process is the key to the mass propagation of plant tissue culture.
5 Acclimatization All explants that have become plantlets (quite large) are washed in running water until clean, then given a fungicide. Planlets are then planted in the media: Fern roots / Moss spaghnum / Burned husk. The planlet is kept in enclosed container or pot to maintain moisture until they become stable.
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