Aseptic_Procedures - Aseptic Procedures Necessity Plants...

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1 Aseptic Procedures Necessity . Plants have certain basic requirements that enable them to live, grow, and reproduce. The same is true for tissues and cells. One of these requirements is a source of energy, which for the intact plant, is light for photosynthesis. Photosynthesis is restricted to cells and tissues that contain chlorophyll. Hence, non-photosynthetic tissues meet their energy requirements by cellular respiration of organic materials. Sucrose, a source, not only of energy but also of carbon, which is required for growth, is supplied to non-photosynthetic tissues via transport systems. Thus, cells throughout the plant body, photosynthetic or not, rely on the energy of respiration to synthesize organic compounds. Excised or cultured plant tissues retain their requirements for specific organic substances. If the tissues in culture require only a general carbon and energy source, a simple carbohydrate such as sucrose may be all that is necessary to be supplied in the culture medium. However, if the requirements are more specific, other organic substances may be necessary; for example, vitamins, amino acids, growth regulators, and so on. If the cultured tissues are unable to synthesize essential substances, these substances must be supplied in the culture medium. Bacteria and fungi, like cultured plant tissues, have many of the same basic requirements for life. However, microorganisms, unlike plant cells, multiple very rapidly, especially in a culture medium containing sugars. Their presence can have a deleterious effect on the plant tissues. Therefore, special techniques, referred to as Aseptic Procedures, must be used to eliminate and prevent microbial contamination. Without the use of aseptic techniques, plant tissue culture would be extremely difficult, if not impossible. Equipment . Although the equipment for plant tissue culture may vary, there are certain basic elements that are essential. These basic items will be described. Transfer Hood . The laboratory room is a major source of contamination. Therefore, the working area must be further confined, preferably to a small, enclosed chamber—the transfer hood. Transfer hoods are generally “box-like” with a front opening. The hood that we are using in this class is known as a laminar flow hood. A blower is used to force air from the top or back to the front of the hood. The air passes through a hepa-filter, which theoretically makes it free of bacteria and fungi. The blower is turned on before the hood is used and remains on while the hood is in operation. If contamination arises, this usually means that the user has not employed proper sterile technique or that the filter needs to be changed.
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