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                PLANT TISSUE CULTURE











     The conventional breeding methods are most widely used for crop improvements. But in certain situations, these methods have to be supplemented with plant tissue culture techniques either to increase their efficiency or to be able to achieve the objective, which is not possible through the conventional methods. One example of each situation would illustrate the point. Production of pure lines or inbreeds involves six to seven generations of selfing. Production of haploids through distant crosses or using pollen, anther or ovary culture, followed by chromosome doubling, reduces this time to two generations. this represents a saving of 4-6 years. The other example is the transfer of a useful bacterial gene say, cry (crystal protein) gene from bacillus thuringienis, into a plant cell and, ultimately, generation of whole plants contain and expressing this gene (transgenic plant). This can be achieved by only by a combination o tissue culture and genetic engineering, non of the conventional breeding approaches can ever produce such a plant.


     The term tissue culture is commonly used in a very wide sense to include in vitro culture of plant cells, tissue as well as organs. But in a strict sense, tissue culture denotes the in-vitro cultivation of plant cells in an unorganized mass, e.g. Callus cultures, another term, cell culture is used for in vitro culture of single or relatively small groups of plants cells, e.g., suspension cultures. But, in general, the term tissue culture is applied to both callus and suspension culture, and cell culture is often used for callus culture as well. When organized structure like root tips, shoot tips, embryo, etc. are cultured in-vitro to obtain their development as organized structures, it is called organ cultures. In this book, plant tissue culture is used in its broad sense to denote aseptic in-vitro culture of plant cell, tissue and organs.







     The in-vitro techniques were developed initially to demonstrate the totipotency of plant cells predicted by haberlandt in 1902. Totipotency is the ability of a plant cell to perform all the function of development, which are charterstics of zygote, i.e. its ability to develop into a complete plant. In 1902, haberlandt reported culture of isolated single palisade cells from leaves in knops salt solution enriched with sucrose. The cells remained silver for up to one month, increased in size, accumulated starch, but failed to divide. Efforts to demonstrate totipotency led to the development of techniques. For cultivation of plant cells under defined conditions. This was made possible by the brilliant contribution from R.J. gautheret in France and p.r. white in U.S.A. during the third and the forth decades of 20th century. Most of modern tissue culture media have been derived from the work of skoog and coworkers during 1950s and 1960s. the first embryo culture although crude, was carried out by hanning in1904, he cultured nearly mature embryo of certain crucifers and grew them to maturity. The technique was utilized by laiubach, in 1925 to recover hybrid progeny from and interspecific cross in linum, subsequently; contributions from several workers have led to be the considerable refinements in of these techniques.


     Haploid plants from pollen grains were first produced by maheshwari and guha in 1964 by culturing anthers of datura. This marked the beginning of anther culture or pollen culture for the production of haploid plants the technique have been further developed by many workers, more notably by j.p. nitch and c. nitch and coworkers. These workers showed that isolated microspores of tobacco produce complete plants.





Tissue culture is a major field of biotechnology witch is completed in a deep description. There are many several points to study tissue culture in briefly.-


      Historical background and terminology used in cell and tissue culture. Basic techniques of cell and tissue culture surface sterilization, aseptic tissue transfer, concept of totipotency. Nutrition requirement of cell in vitro, various types of nutrient media. Somatic embryogenesis and organogenesis in plants. Variability in tissue cultures, somoclonal and other variations, isolations of cells, single cell culture and cloning. Micro propagation and cloning of plants, application of micro propagation in agriculture, horticulture and forestry. Haploid production, various techniques, applications. Production of disease free plants by tissue culture methods. Protoplast isolation and culture, fusion of protoplasts. Somatic hybrids, selection methods, gene expression in somatic hybrid.





  • Enormous opportunities in the study of single cells and group of cells.
  • In the isolation of protoplast.
  • In cell cloning by the plating technique with or without specific treatment, e.g. Mutagens, amino acid analogues etc.
  • Development of cell lines for various types of resistance e.g. Salt and drought and toxin resistant lines.
  • In scale up technology using bioreactors of various types.
  • In the study of nutrition.
  • In secondary product formation, regulation and biosynthesis.
  • In the process of cell division and factors affecting cell division and related process.






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