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Micropropagation (or in vitro propagation) is the most common term used for clonal, true‐to‐type propagation of plants by a variety of tissue, cell and organ culture methods. It implies the aseptic culture of small sections (i.e., explants) of tissues and organs, in closed vessels with defined culture media and under controlled environmental conditions. Micropropagation, in addition to genetic engineering, is at present the most commercially efficient and practically‐oriented plant biotechnology, resulting in rapid generation of a large number of clonal plants of many plant species, which are in many cases also virus‐ or other pathogen‐free. It is now the technical link in the generation of transgenic plants and somatically‐bred plants. Following a brief history and a presentation of the objectives of micropropagation and related applications, we discuss the basic biological principles of micropropagation, including totipotency of plant cells, cell division and callus formation, and the major forms of differentiation and regeneration (axillary bud proliferation, organogenesis, and somatic embryogenesis). The practice of micropropagation is dependent on precise implementation of several technical aspects and developmental/growth stages, including the use of proper explants, asepsis (axenic culture), culture media composition and use of appropriate growth vessels, and the monitoring and control of macro‐ and micro‐environmental conditions within the laboratory and acclimatization facility. We include a review of the design and maintenance of the laboratory and further discuss the advantages and disadvantages of micropropagation and the organizational, economic and financial aspects of commercial micropropagation. In addition, we address critical limiting factors such as recalcitrance, somatic variation and trueness‐to‐type, and contamination. New trends in improving the efficiency of micropropagation conclude this chapter.
Micropropagation (or in vitro propagation) is the most common term used for clonal, true‐to‐type propagation of plants by a variety of tissue, cell and organ culture methods. It implies the aseptic culture of small sections (i.e., explants) of tissues and organs, in closed vessels with defined culture media and under controlled environmental conditions. Micropropagation, in addition to genetic engineering, is at present the most commercially efficient and practically‐oriented plant biotechnology, resulting in rapid generation of a large number of clonal plants of many plant species, which are in many cases also virus‐ or other pathogen‐free. It is now the technical link in the generation of transgenic plants and somatically‐bred plants. Following a brief history and a presentation of the objectives of micropropagation and related applications, we discuss the basic biological principles of micropropagation, including totipotency of plant cells, cell division and callus formation, and the major forms of differentiation and regeneration (axillary bud proliferation, organogenesis, and somatic embryogenesis). The practice of micropropagation is dependent on precise implementation of several technical aspects and developmental/growth stages, including the use of proper explants, asepsis (axenic culture), culture media composition and use of appropriate growth vessels, and the monitoring and control of macro‐ and micro‐environmental conditions within the laboratory and acclimatization facility. We include a review of the design and maintenance of the laboratory and further discuss the advantages and disadvantages of micropropagation and the organizational, economic and financial aspects of commercial micropropagation. In addition, we address critical limiting factors such as recalcitrance, somatic variation and trueness‐to‐type, and contamination. New trends in improving the efficiency of micropropagation conclude this chapter.
Research has revealed that most chlorophyllous explants/plants in vitro have the ability to grow photoautotrophically (without sugar in the culture medium), and that the low or negative net photosynthetic rate of plants in vitro is not due to poor photosynthetic ability, but to the low CO 2 concentration in the air-tight culture vessel during the photoperiod. Moreover, numerous studies have been conducted on improving the in vitro environment and investigating its effects on growth and development of cultures/plantlets on nearly 50 species since the concept of photoautotrophic micropropagation was developed more than two decades ago. These studies indicate that the photoautotrophic growth in vitro of many plant species can be significantly promoted by increasing the CO 2 concentration and light intensity in the vessel, by decreasing the relative humidity in the vessel, and by using a fibrous or porous supporting material with high air porosity instead of gelling agents such as agar. This paper reviews the development and characteristics of photoautotrophic micropropagation systems and the effects of environmental conditions on the growth and development of the plantlets. The commercial applications and the perspective of photoautotrophic micropropagation systems are discussed.
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