A Brief History of Plant Tissue Culture Tissue culture when interpreted into German called Gewebe culture or tissue

culture (UK) or weefsel Kweek or weefsel cultuur (Netherlands). Tissue culture or cultured in vitro is a method to isolate parts of plants such as protoplasm, cells, tissues or organs are completely sterile, grown on artificial media are sterile, in a sterile culture bottle and in aseptic conditions, so that the parts can be memperbayak themselves and regenerate into complete plants. Basic theory used is totipotensi theory written by Schleiden and Schwann that states that the theory totipotensi is part of the living plant has totipotensi, when cultured in appropriate media, will be able to grow and develop into a perfect plant, that can reproduce, multiply normally through seeds or spores. Application of tissue culture in the first place is to plant propagation. Furthermore, the use of tissue culture grow even more is to produce disease-free plants, germplasm collection, improve the properties of plant genetics, production and extraction chemical substances that are useful from the cells - the cells cultured. Many benefits can be obtained from the vegetative propagation of genetic properties can be maintained so as to produce the same plant with its parent. The use of tissue culture techniques started by Gottlieb Haberlandt in 1902 in an attempt cultured hair cells of the monocot plant leaf mesophyll tissue. But the effort failed because the cells do not have cleavage, it is alleged a failure because it does not use growth regulators required for cell division, proliferation and induction of embryos. In 1904, Hannig planting embryos isolated from several plant crucifers. In 1922, Knudson and Robbin separately each conduct business and culture of planting orchid root tips. After the 1920s, the discovery and development of tissue culture techniques continues. The following table shows the historical development of the field of plant tissue culture which was adapted from various sources. Tissue culture techniques require certain conditions to be fulfilled in its implementation. Principal requirement is the implementation of tissue culture laboratory with all amenities. The laboratory must provide the tools work, means of supporting the creation of controlled aseptic conditions and basic amenities like, water electricity and fuel. Implementation requires tissue culture is also qualified software. In making the implementation of tissue culture, implementers must have a background of certain basic sciences of botany, plant physiology ZPT, chemistry and physics are adequate. Implementing is engaged in work directly related to basic sciences. Implementing will deal with various kinds of chemicals, process plant physiology (biochemistry and physics) and various kinds of analytic work. Sometimes background knowledge of microbiology, cytology and histology. Executor is also required in terms of work skills; perseverance and patience are high and have to work intensively. Works include tissue culture: media preparation, isolation of planting material, sterilization of explants, explants inoculation, acclimatization and transfer the business results of tissue culture plants to the field. Implementers have to work carefully and seriously, because each stage of the job requires the handling of its own with its own knowledge base. The characters that play a role in the history of the commencement of tissue culture of knowledge include: y The use of tissue culture techniques started by Gottlieb Haberlandt in 1902 in an attempt cultured hair cells of the monocot plant leaf mesophyll tissue. But the effort failed because the cells do not have cleavage. Allegedly his failure was due to disuse of plant growth regulators required for cell division, proliferation and induction of embryos. y 1904, Hannig perform embryo culture on cruciferous plants. y Knudson successfully sprouted orchid in vitro in the year 1922, the same year Robbins cultured root tips in vitro. y In 1924 Blumenthal & Meyer: The formation of callus from carrot root explants y In 1929 Laibach & Hered: Culture of embryos to overcome the incompatibility in plants Linum spp. y In 1936 LaRue: Culture of embryos in some gymnosperms plant y Gautheret, nobecourt and White who found Auxin and has managed to cultivate callus in 1939. y Skoog et al. have found cytokines and the first to succeed in doing tissue culture in 1939. y In 1940 Gautheret cambium does tissue culture in vitro on Ulmus plants to study the formation of adventitious buds.

y 1941 The use of coconut water to mix media in cultured Datura by Van Overbeek. y The first adventitious shoot formation in tobacco culture in vitro by Skoog in 1944. y New in 1946, the first complete plant can be produced from shoot tip explants in culture and Tropaeolum Lupinus by Ball. y In 1948 Skoog and Tsui: Formation of adventitious shoots and roots of tobacco. y In 1949 Nitsch: in vitro culture of fruit trees. y In 1950, Ball tried to plant crops Sequoia sempervirens callus tissue and may result in organs. y Muir managed to grow a complete plant from a single cell culture in 1954. y In 1955 Miller et al. finding kinetin which can stimulate cell division. y Production of the first haploid plants generated by Guha in 1964. y Laminar flow is used first in the late '60s. y Power tries to make the union (fusion) Protoplast first time in 1970. y New in 1971 produced complete plants from protoplast by explants Takebe. y To obtain disease resistant plants, Larkin in 1981 to conduct research first somaclonal variation. y One way to get kultuvar excel is to do the transformation. The first cell transformation by Horch in 1984. y The first plant transformation performed by the IPTC in 1986. y Transformation of wheat by Vasil in 1992. y In 1996 the first release of genetically transformed plants. Platform for Tissue Culture The foundation of tissue culture based on three basic capabilities of the plant, namely: 1. Totipotensi is the potential or ability of a cell to grow and develop into a whole plant if properly stimulated and appropriate hearing. Implications of totipotensi are that all the information about the growth and development of an organism found in the cell. Although in theory the whole cell is totipotensi, but the best success is a cell expressing a meristematic. 2. Rediferensiasi is the ability of mature cells (mature) back into the condition and meristematic and develop from one point of new growth, followed by rediferensiasi who became an organ capable of performing the new reorganization. 3. Competencies describe the potential of endogenous cells or tissues to grow and thrive in one particular path. Cantohnya embrioagenikali competent cells are the ability to develop into an embryo functional full. In contrast is a non-competent or morfogenetikali not have the ability. The development of tissue culture in Indonesia The development of tissue culture in Indonesia was very slow, even almost said roads in place when compared with other countries, it is no wonder that orchid seeds imported in the form of 'flasks' could flood the nursery, orchid nurseries in our country. In addition to the technology gap in the line of academics, research institutions, public and orchid lovers, one of the causes of this technology is becoming very slow progress is due to a perception that the required investment is 'very expensive' to establish a tissue culture lab, and only fit or 'feasible' for the company. Indonesia has a remarkable biodiversity, one of which is the orchid, an estimated 5000 species of orchid species spread over forest area of Indonesia. The potential is very valuable for developers and lovers of orchids in Indonesia, particularly the genetic potential to produce a hybrid orchid which has a high commercial value. This potential will be meaningless when logging and massive exploitation of our forests occur, not to mention the blatant theft or "hidden" under the pretext of cooperation and contribution of research both by our society and foreigners. Meanwhile, only a small party that is able to perform the development and utilization of orchid species, especially those related to tissue culture technology. No doubt that the best method to date in the preservation and propagation of orchids is by tissue culture, because through tissue culture many things that can be compared with the conventional method. In principle, tissue culture lab can be simplified by modifying the equipment and materials used, so it is possible to tissue culture such as 'home industry'. This can be seen in the group of farmers seed orchid culture' in Malang that has been so many.

Tissue culture techniques The main principle of this technique is multiplication plant tissue culture by using vegetative parts of plants using artificial media is done in a sterile place. Tissue culture method was developed to help reproduce the plant, especially for plants that are difficult bred generative. Seedlings produced from tissue culture has several advantages, including: having the nature of which is identical to the parent, can be propagated in large numbers so as not to need a large place, able to produce seeds in large quantities in a short time, better health and quality of seeds guaranteed, seedlings grew faster speed compared with conventional propagation. Steps being taken in plant propagation by tissue culture techniques are: 1) Making media 2) Initiation 3) Sterilization 4) Multiplication 5) Rooting 6) Acclimatization Media is a key factor in the multiplication by tissue culture. Composition of media used depends on the type of plants to be propagated. Media used are usually composed of mineral salts, vitamins, and hormones. In addition, also required additional materials such as agar, sugar, and others. Growth regulators (hormones) that is added will also vary, both type and number, depending on the purpose of tissue culture is performed. Media that had already been placed in test tubes or glass bottles. The medium used must also be sterilized by heating it with an autoclave. Initiation is a collection of explants from the plant to be cultured. The plant is often used for tissue culture is the bud. Sterilization is that all activity in tissue culture should be performed in a sterile, in the laminar flow and using the tools that are also sterile. Sterilization was also made to the equipment; the use of ethanol is sprayed evenly on the equipment used. Technicians who perform well tissue culture must be sterile. Multiplication is the activity of candidate plants reproduce by growing explants on the media. This activity is performed in a laminar flow to avoid any contamination that caused the failure of explants growth. Test tube that has been planted explants placed on the shelves and placed in a sterile room temperature. Rooting is the phase in which the explants would indicate the presence of root growth that marked that the process is carried out tissue culture started going well. Observations were made every day to see the growth and development of roots and to see the existence of contamination by bacteria or fungi. Contaminated explants would show symptoms such as white or blue (due to fungus) or rot (caused by bacteria). Acclimatization is an activity to move out of the room aseptic explants into beds. The transfer is done carefully and gradually, by providing containment. Lid is used to protect seedlings from outside air and pest and disease because the results of tissue culture seedlings are very susceptible to pests and diseases outside air. Once the seeds are able to adapt to his new environment, the lid is released gradually and maintenance of seedlings performed in the same manner with the maintenance of generative seed. Excellence is of interest to seed producers to begin to develop a business culture of this tissue. Already, there is some forest plants bred with tissue culture techniques, such as: teak, sengon, acacia, etc. Results of tissue culture seedlings grown in some areas showed good growth, even teak tissue culture results are often called the golden teak can be harvested within a relatively shorter compared with teak plants originating from the generative seed, regardless of the quality of wood that has not tested in Indonesia. This is very profitable business because it will get quicker results. Advantages of the Use of Tissue Culture y Procurement of seedlings did not depend on the season. y Seeds can be produced in large quantities with a relatively faster (than a bud eye that had a response within 1 year can be produced at least 10,000 plantlets / seed). y Seedlings produced uniform. y Disease-free seedlings produced (using a particular organ). y Seedling transportation costs are relatively less expensive and easier. y In the process of nurseries are free of pests, diseases, and other environmental flogging.