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Basic Techniques of Plant Tissue Cultures

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A description of techniques used to successfully grow tissue cultures, including several videos of specific steps.

Asceptic Technology

Yang Yen
College of Agriculture and Biological Sciences at South Dakota State University, USA

Patricia Hain
Department of Agronomy and Horticulture at University of Nebraska-Lincoln, USA

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Contaminated Culture

Plant tissue culture is the in vitro growing of parts of a plant such as organs, tissues or cells on an artificial nutritional medium. This culture medium is also an excellent resource for the growth of contaminates such as fungus and bacteria, and the growth of the contaminates easily outpaces that of the cultured plant cells. Therefore, general procedures of microbiology are applied to avoid contamination. Basically, anything that may have the chance to make a direct contact with the culture medium and/or explants has to be sterilized and manipulations should be designed such that there is no direct link between the culture and its nonsterile surroundings.

Good Culture.
Good Culture

Tissue culture practice usually uses three levels of containment to preserve sterile conditions during culture manipulations: using sterilized medium, tools, containers and explants; working under a sterilized environment; and practicing sterilizing techniques. Autoclaving, surface treatment and filtration are three means to achieve sterilization of the tools and materials used.

Development of this lesson was supported in part by Cooperative State Research, Education, & Extension Service, U.S. Dept of Agriculture under Agreement Number 98-EATP-1-0403 administered by Cornell University and the American Distance Education Consortium (ADEC).  Any opinions, findings, conclusions or recommendations expressed in this publication are those of the author(s) and do not necessarily reflect the view of the U.S. Department of Agriculture.


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