Biotechnology: Scope and Importance PDF

CHA PTER Biotechnology : Scope and Importance A. WHAT IS BIOTECHNOLOGY ? 1917 a Hungarian Engineer, Karl Ereky coined the term In biotechnology to describe a process for large scale production of pigs....

CHA PTER Biotechnology : Scope and Importance A. WHAT IS BIOTECHNOLOGY ? 1917 a Hungarian Engineer, Karl Ereky coined the term In biotechnology to describe a process for large scale production of pigs. According to him all types of work are biotechnology by which products are produced from raw materials using living organisms. During the end of 20th century biotechnology emerged as a new discipline of biology integrating with technology; but the route of biotechnology lies in biology. There was no sudden sprout of this discipline, but some of the methods for production of products were developed centuries back. Therefore, biotechnology is concerned with exploitation of biological components for production of useful products. Biotechnology is defined by different organisations in different ways. It has been broadly defined as, the barley and a top-fermenting yeast (Saccharomyces development and utilization of biological cerevisiae) with water at about 60-75°F processes, forms and systems for obtaining maximum benefits to man and other forms of life". Biotechnology is the science of applied biological process" (Biotechnology : A Dutch Perspective, 1981). Following are some of the definitions given by other organisations : 1 Copyrgtited matoriat  2 A Textbook of Biotechnology Biotechnology is the application of scientific and engineering principles to the processing of materials by biological agents to provide goods and service [The Organisation for Economic Cooperation and Development (OECD), 1981]. The integrated use of biochemistry, microbiology and engineering sciences in order to achieve technological application of the capabilities of microorganisms, cultured tissue, cells, and parts their of [The European Federation of Biotechnology (EFB), 1981; O'Sullivan, 1981]. The application of biochemistry, biology, microbiology and chemical engineering to industrial process and products and on environment [International Union of Pure and Applied Chemistry (IUPAC), 1981.] Biotechnology is the controlled use ofbiological agents such as microorganisms or cellular components for beneficial use" U.S. National Science Federation). In the definition given by OECD, "scientific and engineering principles" refer to microbiology, genetics, biochemistry, etc. and biological agents means microorganisms, enzymes, plant and animal cells. The meaning of these three definitions and others given by many organisations are more or less similar. A unified definition of genetic engineering has been given by Smith (1996) as "theformation of new combinations of heritable material by the insertion of nucleic acid molecules produced by whatever means outside the cell, into any virus, bacterial plasmid or other vector system so as to allow their incorporation into a host organism in which they do not naturally occur but in which they are capable of continued propagation". B. HISTORY OF BIOTECHNOLOGY If we trace the origin ofbiotechnology, it is as old as human civilization. Development of biotechnology can be studied considering its growth that occurred in two phases: () the traditional (old) biotechnology, and (ii) the new (modern) biotechnology. 1. Traditional Biotechnology Really the traditional biotechnology is the kitch en technology developed by our ancestors using the fermenting bacteria. Kitchen technology is as old as human civilisation. During vedic period (5000-7000 BC), Aryans had been performing daily Agihotra or Yajna. One of the materials used in Yajna is animal fat (i.e. ghee) which is a fermented product of milk. Similarly, the divine 'soma' (a fermented microbial product used as beverage) had been offered to God. Summarians and Babylonians (6000 BC) were drink ing the beer. Egyptians were baking leavened bread by 4000BC. Preparation of curd, ghee, wine beer, vinegar, etc. was the kitchen technology. In spite of alldevelopment, preparation of curd, ghee, vinegar, alcoholic beverages, jalebi, idli, dosa, have become an art of the kitchen of all Indians (Table 1.1). Churning of yogurt in eathen pot makes cultured butter in India Copynghted matenat  Biotechnology : Scope and Importance 3 > Table 1.1 Traditionalfermented foods prepared in different parts of India Types of Substrate Regions Quality and uses foods Ambali Millet and rice South Steamed cake used in snack Bhatura Wheat flour (maida) North Flat fried, lavened bread used with chhola Chhurpi Milk Himalaya Cheese like, mild sour, soft mass, used as curry Dosa Rice and black gram South Spongy, shallow fried, used as staple food Dahi Milk North Sour, thick gel with whey Dhokla Bengal-gram West Spongy cake used as snack Gundrum Leafy vegetable Himalaya Sun-dried, sour taste, used as-soup or pickles Idli Rice and black gram South Steamed spongy cake used in breakfast Jalebi Wheat flour (maida) North Crispy, deep-fried used as sweet confectionery Khaman Bengal gram West Spongy cake, used in breakfast Khalpi Cucumber Himalaya Sour pickles Mesu Bamboo shot Himalaya Sour pickles Mishti dahi Milk East Thick sweet gel Nan Wheat flour (maida) North Leavened flat baked bread used as staple food Papad Black gram flour North Circular wafers used as snack (besan) Paneer Milk North Soft cheese used as fried curry Rabadi Mixture of butter-milk- West Cooked paste used as staple food wheat/pearlmillet/barley Srikhand Milk West Concentrated sweetened preparation Sinki Radish tap root Himalaya Sun-dried sour soup pickles Tari Date palm East Sweet alcoholic beverage Tharra Mahua North Sweet alcoholic beverage obtained through distillation Vadai (wada) Black gram North Deep fired cake used as snack Wari Black gram North Spongy cake used as snack The traditional biotechnology refers to the conventional technology which have been used for many centuries. Beer, wine, cheese and many foods have been produced using traditional biotechnol ogy. Thus, the traditional biotechnology includes the process that are based on the natural capabilities of microorganisms. The traditional biotechnology has established a huge and expanding world market. In monetary term, it representsa major part of allbiotechnology financial profits. In India who can forget the story of Makhan Chori (butter stealing) by Lord Sri Krishna during Mahabharat period ? Butter would have been produced following the same kitchen art. Besides breeding of strong productive animals, selection of desirable seeds for enhanced crop production has been the part of human activity since the time immemorial. The Egyptians (about 2000 BC) used to prepare vinegar from crushed dates by keeping for longer time. But the crushed dates produce intox icants at first. In Egypt, Mesopotamia and Palestine (about 1500 BC) the art of production ofwine from crushed grapes, and beer from germinated cereals (malt) using a bread leaven (a mass of yeast) was established. In Indian 4yurved, production of Asava' and 'Arista' using different substrates, Copyrignted materal  4 A Textbook of Biotechnology and flowers of mahua (Madhuca indica) or dhataki (Wodfordia fructicosa) has been well characterised till today since vedic period. In these methods various substrates are transformed into a number of products. Hence, the odour, colour and taste of final products are changed. Moreover, use of salt for preservation of various food has been earlier developed even in Europe. Still preservation methodology of the mummies of Egypt are noteworthy. Possibly, mummification would have been done through dehydration of dead body followed by use of mixture of salts largely sodium carbon ate. Thus, the traditional biotechnology was an art rather than a SCience. Role of Microorganisms in Fermentation: The causes of fermentation could be discovered after observing microorganisms using a microscope by Antony van Leeuwenhoek (1673-1723) at Delft (Holland). During 18th century a significant contribution was made by the chemists on the process and products of fer mentation. In 1757, it was demonstrated that a milky precipitate could be obtained when the gas evolved from fermentation was passed through the lime water. It was called lime water test. A Antony Van micrOSCope. similar gas comes out from burning of charcoal. Henry Cavendish demonstrated that the gas evolved from brown sugar in water treated with yeast was absorbed by sodium hydroxide solution. Schele and J Priestley (1772-1774) confirmed the identity of oxygen gas present in air. Using analytical technique for carbon estimation, Antoine Lavoisier gave the chemical basis of alcoholic fermentation. A French man, Nichola Appert (1810) described the method of food preservation. In the same year Peter Durand also gave the use of tin container for food preservation. It was done by putting an air-tight vessel containing food material in the boiling water. It increased the importance of canning industry. Lack of oxygen in such a closed and heated vessel was reported by Gay Lussac. He concluded that oxygen was required for initiation of alcoholic fermentation, but not for further progress of fer mentation. After 1830, Charles B. Astier gave the concept that air is the carrier of allkinds of germs. In 1837, Theodore Schwann after a series of experimentation demonstrated that the develop ment of the fungus (sugar fungus) on fruit juice causes fermentation'. He was the first to observe and describe the yeast in growing process. Charles Cag niard-Latour (1838) observed yeast budding using a microscope allowing 300-400 power magnification. Justus von Liebig (1839), a well known chem ist, proclaimed that all the activity of yeast cells was the result of chemical and physical reactions going on in the mediunm. The study of microbiology was started since the first report of Louis Pasteur (1857) on lactic acid fermentation from sugar. He isolated the micro organisms (lactic yeast) that were associated with lactic acid and formed curd. The cells of lactic yeast were smaller than that of beer yeast. Lactic acid pro duction got increased when he added chalk powder Louis Pasteur Copynghted materat  Biotechnology : Scope and Importance 5 to fermentation medium. Pasteur showed the presence of lactic acid in curd by using a polarimeter. In 1860, Pasteur provided a detailed report on the use of synthetic medium for microbiological studies. He concluded that: (i) fermentation is carried out anaerobically by the living cells. (ii) the yeast increased the weight with increase in C and N contents of overall batch during alcoholic fermentation in synthetic medium. The increase in yeast protein in synthetic me dium was accompanied by a related decrease in the ammonium nitrogen in the medium. (ii) fermentation of sugar was required for multiplication of yeast cells. (iv) similar phenomenon occurred in fermentation of lactic acid, tartaric acid, butyric acid, etc. (v) because of not using pure culture, some of the fermentation processes were stopped. (vi) growth and physiology of yeast differ when they are grown under aerobic and anaerobic conditions (it was later on called as Pasteur effect'). Under anaerobic conditions a large amount of sugar was converted into alcohol, while under aerobic conditions large amount of sugar was converted into yeast cell mass. Pasteur suggested that high percentage of microbial population is killed by heating the juice at 62.8° C(145° F) which is now called as Pasteurization. Robert Koch (1881) gave the method for establishing relationship of apathogen with a disease which is known as Koch's postulates' or 'pathogenicity test'. Following this technique he proved that the anthrax disease is caused by Bacillus anthracis. Edward Buchner (1897) was the first to demonstrate enzymatically-mediated fermentation reactions. He showed that cell-free yeast juice mixed with concentrated sugar solution evolved upon incubation the carbon dioxide and produced ethyl alcohol. Similar products were also pro duced in aqueous solution of the other sugars such as glucose, sucrose, fructose and maltose. Buchner called the dissolved substance in juice responsible for sugar fermentation as 'zymase'. This work showed the improved techniques of fermentation. Fermentation could also be demonstrated in test tubes without a living organism. The work of chemistry had more role in understanding the fermentation phenom enon than the microbiology. Edward Jenner During 1890s, Hans Buchner and Martin Hahn developed more effective method of getting cell-free extracts after disrupting the microbial cells. They ground the microbial cells with quartz and adding Kieselguhr (to get sufficient consistency in the resulting paste). Thus, sucrose fermentation using cell-free extracts from yeast cells (obtained by the above method) was demonstrated. Discovery of viruses and their role in disease was possible when Charles Chamberland (1884) constructed a porcelain bacterial filter. Edward Jenner (1798) used vaccination by taking out liquid material from cowpox lesions and introducing into people having smallpox. But Pateur and Chamber land developed an attenuated anthrax vaccine against anthrax disease. After the discovery of toxins produced by Corynebacterium diphtheriae (causes diphtheria in human) its antitoxin was developed by Emil von Behring (1890) and Shibasaburo Kitasato. He injected the inactivated toxin into rabbit that induced to produce antitoxin in the blood. The antitoxin inactivated the toxin and protected against the disease. Similarly, tetanus antitoxin was developed by Behring. Copyrg ted matoral  6 A Textbook of Biotechnology 2. Modern Biotechnology The two major features of technology differentiates the moden biotechnology from the classical biotechnology: (i) capability of science to change the genetic material for getting new products for specific requirement through recombinant DNA technology, and (i) ownership of technology and its socio-political impact. Now the conventional industries, pharmaceutical industries, agro-industries, etc. are focusing their attention to produce biotechnology-based products. (a) Emergence of Modern Biotechnology: The new or modern biotechnology embraces all methods of genetic modification by recombinant DNA and cell fusion technologies. It also includes the modern developments of tradi tional biotechnological processes. The newW aspects of biotechnology founded in recent advancement of modern biology, genetic engineering and fermentation process technolo gy are now increasingly find wide industrial application. But the rate ofapplication will depend on: (i) ad equate investment by the industries, (ii) improved system of biological patenting, (i) marketing skill, (iv) Canning process. economics of the new methods, and (v) public perception about the biotechnology products. With the end of 19th century, the traditional biotechnology associated with fermentation was gradually industrialised. This resulted in gradual growth of industries producing beer, whisky, wine, rum, canned food, etc. In 1920, for the first time, the Leeds City Council (U. K.) established the Institute of Biotech nology. In the late 1960s, OECD was set up to promote policies for sound economic growth of the member countries. In 1978, the European Federation of Biotechnology was set up. In 20th century, biotechnology brought industries and agriculture together. During World war Ifermentation processes were developed which produced the acetone from starch and paint solvent from automobile industries. During World war II the antibiotic penicillin was discovered. Manufacture of penicillin shifted the biotechnological focus towards pharmaceuticals. Linking the fermentation with biochemistry, bioprocess, chemical engineering and instrument designing helped substantially in the progress of industries. During Gulf War (1991) the work on microorganisms dominated for the preparation of biological warfare, antibiotics and fermentation process. Suspected preparation of biological and chemical warfare led to US attack on Iraq in 2003. After the discovery of double helix DNA by Watson and Crick (1953), Werner Arber (1971) discovered a special enzyme in bacteria which he called the restriction enzymes. These enzymes can cut the DNA strand and generate frag ments. The cut ends of two fragments are single stranded sticky ends because the single stranded ends having identi cal base pairs can re-join. In 1973, S. Cohen and H. Boyer removed a specific gene from a bacterium and inserted into another bacterium using restriction enzymes. This discovery A research scientist preparing marked the start of recombinant DNA technology or genetic plasmid DNA engineering. In 1976, Baltimore and Baltimore successfully Copyigtited matorial  Biotechnology : Scope and Importance 7 transferred human growth hormone gene into a rabbit. In 1978, the European Federation of Biotech nology was created. In 1978, a U.S. company Genctech' used genetic engineering technique to produce human insulin in E. coli. In 1980, trials of new hormone was conducted in the U.S.A., France, Japan, and the United Kingdom. The US Food and Drug Administration gave marketing approval to Humulin' i.e. human insulin made by Eli Lilly (U.S.A) by the end of 1982. Another hormone 'somatotropin was produced on industrial scale. In 1993, the first genetically engineered tomatoes, FlavrSavr, were sold in market. In 1996, the first clone lamb Dolly' was borne successfully by the efforts of scientists of Scotland. Thereafter, several cloned animals were produced. In 2001, the sequence of the Human Genome was published in Nature and Science. Human Ge nome Project was completed by March 2003. In 2002, a designer baby was born to cure the genetic disease of her elder sister. On December 27, 2002, a claim was made for the birth of a clone baby Eve' by the scientists of "Human Cloning Society', the Clonaid of France. In February 2005, in the meeting of United Nations many countries opposed gene cloning in humans, while a few countries supported with request to allow for the sake of research only. In May 2005, scientists in South Korea have used a method called therapeutic cloning to pro duce stem cell lines. These are genetic matches to patients. Such stem cell lines could be used for disease research. The U.S.A. condemned this approach. In this method human embryos were produced through cloning (as done for Dolly) and stems cells were obtained from blastocyst. The excised stem cells could be grown in vitro and used further. It is, therefore, interesting that the scientists are engaged in doing the most challenging task of mass production of growth hormones, insulin, vaccines, immunogenic proteins and polypeptides, gene therapy, biofertilizers, biopesticides, producing disease and stress-resistant plants, biomass, enzymes, antibiotics, acids, fuels, etc. Many biochemical companies such as National Pituitary Agency (U.S.A.), E. Lilly (U.S.A.), Kabi Vitram A B (Sweden), Genetech Co. (U.S.A.), Biogen (Switzerland), etc. are producing/trying to produce some of the above products by using genetic engineering techniques. Many Nobel Laureates including Dr. Har Govind Khorana, are associated with these companies. The area of main interest, at present, where such works are being done throughout the world are given in Table 1.2. Many international agencies are making efforts to solve the national and international problems via. collaborative works. Various programmes including molecular and environmental engineering have been taken up by the European Communities with frame work of their collaborative activities. For training purposes, under several biotechnological Genetically engineered human insulin programmes, centres have been established, available in large scale. for example, the International Centre for Cooperative Research and Training in Microbial Engineering (Japan) and Institute of Biotechnological Studies (U.K.). Many countries have developed collaborative networks on several aspects such as Regional Microbiology Network for South-East Asia" (supported by Japan and UNESCO). "Microbiological Resource Centres" (MICRCENS) [Supported by UNESCO]; United Nations Environmental Programmes (UNEP), International Cell Research Organisation (1CRO). These two organisations have participations of groups in many countries. Copyngtited material  A Textbook of Biotechnology > Table 1.2 Area of biotechnology. Area of interest Products 1. Recombinant DNA technology Fine chemicals, enzymes, vaccines, growth (genetic engineering) hormones, antibiotics, interferon. 2. Treatment and utilization of bio Single cellprotein, mycoprotein, alcohol materials (biomass) and biofuels. 3. Plant and animal cell culture Fine chemicals (alkaloids, essential oils, dyes, steroids), somatic embryos, encapsulated seeds, interferon, monoclonal antibodies. 4. Nitrogen fixation Microbial inoculants (biofertilizers) 5. Biofuels (bioenergy) Hydrogen (via photolysis), alcohols (from biomass), methane (biogas produced from wastes and aquatic weeds). 6. Enzymes (biocatalysts) Fine chemicals, food processing, biosensor, chemotherapy. 7. Fermentation Acids, enzymes, alcohols, antibiotics, fine chemicals, vitamins, toxins (biopesticides). 8. Process engineering Effiuent, water recycling, product extraction, novel reactor, harvesting. (b) Biotechnology as an Interdisciplinary Area: Biotechnology is not a sudden discovery, rather a coming of age of atechnology that was initiated several decades ago. By the middle of 20th century there had been a tremendous growth in the area of chemistry, physics and biology. Further knowledge of each branch has been advanced substantially. Multidisciplinary strategies were made for the solution of various problems. A novel spectrum of investigation occurred through the true interdisciplinary synthesis. This led to the evolution of biotechnology which is an outcome of integrated effort ofbiology with technology, the root of which lies in biological science (Fig 1.1). The key difference between biology and biotechnology is their scale of operation. Usually the biologist works in the range of nanograms to milligrams. Biotechnologists working on the production of vaccine may be satisfied with milligram yields, but many other projects aims at kilograms or tonnes. Thus, the main objective of biotechnologists consists of scaling-up the biological processes (Smith, 1996). The main discipline ofbiology are microbiology, biochem istry. genetics, molecular biology, immunology, cell and tissue Biotechnologist lsolating the gene for protein X. culture. However, on the engineering side it includes chemical and biochemical engineering such as large scale cultivation of microbes and cells, their upstream and down stream process, etc. These processes can be separated into five major operations: () strain selection and improvement, (ii) mass culture, (iii) optimisation of cell responses, (iv)process operation, and (v) down stream processing (i.e. product recovery). Many areas ofbiotechnology have arisen through the interaction between various parts ofbiology and engineering, biochemistry, biophysics, cellbiology, colloid chemistry, embryology, ecology, genetics, immunology, molecular biology, medical chemistry, pharmacology, polymer chemistry, Copyngited matorial  Biotechnology : Scope and Importance 9 Biomolecular engineering feed technology 3 Plant/animal cel culture l Wutilzaon aste technology technolgy Biomining teEcnhvirlongmyental Recombinant DNA Fermentation technology technology Biotechnology Biochemicalengineering P h y s i o l o&g b yi o c h e m i s t r y Maths & computer science Fig. 1.1. A Biotechnology tree. thermo-chemistry and virology. The modern biotechnology has developed several technologies extracting the basic knowledge from biology. (c) Global Impact and Current Excitement of Biotechnology: Each and every organism performs its function within its optimum limits. The excitement about the modern biotechnology Copyrighted mater1al  10 A Textbook of Biotechnology is that the scientific methods (such as genetic engineering) have enhanced the natural capabilities of natural production of organisms. What a miracle is that a mouse of the size of rabbit can be produced ? Bacteria like E. coli are producing mammalian hormons such as insulin, somatostatin, somatotropin, etc. Yeast cells have been genetically manipulated to produce vaccine against hepatitis B virus (hepatitis disease). Myeloma cells (cancerous cells of bonemarrow) and B-cells of immunised mice were hybridised to produce hybrid cells that consisted the characteristics of both the cells which were cell division and antibody production. Now the hybrid cells (hybridomas) are being used for production of monoclonal antibodies. In 1982, interferons (a, B and y) were produced by genetically engineered E. coli cells. Techniques have been developed to produce rare and medicinally valuable molecules to change hereditary traits of plants and animals, to diagnose diseases, to produce useful chemicals and, to clean up and restore the environment. In this way biotechnology has great impact in the fields of health, food/agriculture and environmental protection. Due to rapid development the present situation that there is no difference between pharmaceutical firms and biotechnology industry. However, approved products in the pipeline and renewed public confidence made it one of the most promising areas of economic growth in future. India offers a huge market for the products as well as cheap manufacturing base for export (Padh, 1996). Following are some of the arcas where biotechnology has done the best. () Health care : The maximum benefits of biotechnology has been utilised by health care. Biotechnology derived proteins and polypeptides form the new class of potential drugs. For example, insulin was primarily extracted from slaughter animals. Since 1982, human insulin (Humulin) has been produced by microorganisms in fermenters. Similarly, hepatitis B vaccines viz., Recombivax HB® (from Merk), Guni (from Shantha Biotechnics Ltd, Hyderabad), Shanvac® (Biological E. Laboratory), etc. are the genetically engineered vaccines produced biotechnologically. Since 1987, the number of biotechnology-derived new protein drugs has surpassed the new chemical drugs. Table 1.3 shows some of the important products produced through genetically modified organisms. With the advancement of gene manipulation in organisms the science have led to a new revo lution in biology which is called gene revolution. Obviously, it is a third revolution in the science after industrial revolution and computer revolution. Thus the roots of today's biotechnology lies in chemistry, physics and biology. Table 1.3. Example of some therapeutic products produced through recombinant DNA technology. Products Application Interferon Cancer and viral infection Human urokinase (tPA) Plasminogen activator used in vascular disorder Insulin Treatment of diabetes Human factor IV Clotting factor for haemophilia Lympholines Auto-immune functioning Serum albumin In surgery Attenuated pseudorabies virus antigen Vaccine against rabies Tissue plasminogen activator In treatment of heart attack Somatostatin Treatment of human growth disorder Copyrighted material  Biotechnology : Scope and Importance 11 There are about 35 biotechnology-derived therapeutics and vaccines approved by the USFDA alone for medical use, and more than 500 drugs and vaccines to reach in market (Table 1.4). Similarly, about 600 biotechnology diagnostics are worldwide available in clinical practices (Table 1.5). About 130 gene therapy protocols have been approved by the US authorities. India relies on imports of many immunodiagnostic kits. Table 1.4. Biotechnology-derived drug products (Source : H. Padh, 1996) Products Manufacturers Human insulin Eli Lilly, Novo Nordisk, Hoechst Growth factors Eli Lilly, Novo Nordisk, Genetech, Pharmacia Blood factors Amgen, J & J, Sankyo, Chugai, Sandoz, Immunex Interferon Roche, Wellcome, Daiichi, Sumitomo Monoclonal antibodies J& J, Cytogen Vaccines Smithkline, Merk, Shionogi Table 1.5. Approved biotech diagonistics. (Source :H. Padh, 1996) Type Infectious Tumour Analyte Blood Total disease marker & drug SCreening Monoclonal antibody 127 2 433 571 DNA probe 42 0 11 55 Recombinant DNA 11 1 13 Total 180 3 445 637 (ii) Agriculture : Biotechnology is making new ground in the food/agriculture area. Current public debate about BSTC, bovine somatotropin (a hormone administered to cows to increase milk production) typifies an example of biotechnology product testing public acceptance. Similarly, the FlavrSavrTM tomato (produced by transgenic plants engineered by antisense technology to preserve flavour, texture and quality) is a new breed of value added foods. Food biotechnology offers valuable and viable alternative to food problems, and a solution to nutritionally influenced diseases such as diabetes, hypertension, cancer, heart diseases, arthritis, etc. A transgenic Golden rice' has been produced by introducing three genes for the production of vitamin A in Taipei' rice. Several insect resistant transgenic Bt plants have been produced by incorporating insecticidal toxin producing Bt gene of Bacillus thuringiensis into the desired plants. A transgenic cotton named Ingaurd' was released in Australia which contained Bt gene and provided resistance against insects. We can recall the Bt cotton prepared for Andhra Pradesh but sown in Gujarat in 2001 which raised debate throughout the country. Biopesticides are coming to the market and their sales are increasing. Molecular Pharming is a new concept where therapeutic drugs are produced in farm animals, for example, therapeutic proteins secreted in goat milk. There are about a dozen companies that produce lactoferrin, tPA, haemoglobin, melanin and interleukins in cows, goat and pigs. However, it is not surprising that vegetables producing vaccines, insulin, interferon and growth hormones would be available in market in 21st century, beside, human clones and several other miracles. (iii) Human Genome Project (HGP): The major landmark in human history is the human genome sequence. The HGP is an international research programme. Almost the whole human genome has been sequenced and chromosome map has been developed in various laboratories world-wide through Gopyrighted materal  12 A Textbook of Biotechnology co-ordinated efforts. Human chromosome mapping was completed by March 2003. There are about 33,000 functional genes in human. More than 97% genes are non-functional. They do not encode any polypeptide chain. Objectives of human genome project are to: (i) construct the detailed genetic and physical map of human genome, (ii) determine the complete nucleotide sequence of human DNA, (ii) store information in database, (iv) locate the estimated 50,000-100,000 genes within the human genome, (v) address the ethical, legal and social issues (ELSI) that may arise from the project, and (vi) perform similar analysis on the genomes of several other organisms. (iv) Environment : The natural biodegradability of pollutants present in environment has increased with the use of biotechnology. The bioremediation technologies have been found successful to combat the pollution problems (see Chapter 25, Environmental biotechnology). Bioremediation is the use of microorganisms to detoxify pollutants, present in the environment usually as soil or water sediments. The pollutants cause several health problems. Microorganisms which show potential to degradation of oil, pesticides and fertilizers belong to the genera of bacteria Pseudomonas, Micrococcus, Bacillus, and fungi Candida, Cladosporium, Torulopsis, Trichoderma, etc. (v) Genomics and Proteomics: Computer-based study and designing of genome is called ge nomics. Genomics deals with sequencing of thecomplete genome of aparticular organism. Similarly, study of proteins present on genome using computer is called proteomics. The proteomics can be defined as the study of all the proteins present in genome of an organism. With the help of proteomics and genomics the new molecules that can interact with the other partners could be identified. This gives us deep insight into biological pathway. Now the whole genome is available to biologists for scrutiny. There may be new small molecules as potential drug candidates. Therefore, interaction of some molecules can be studied in greater detail. The 33,000 genes of human beings are on a microchip. It has helped to design specific drugs for genetic diseases, for which there is no cure so far. For example, a specific gene (Her-2 Neu) over-expresses in breast cancer patients. A designed drug (Herceptin) is good for treatment of breast cancer. Thus, the field of genomics has helped the growth of pharmacologic, toxicologic and protein studies on animals, therefore, the new areas are called pharmacogenomics, toxicogenomics and proteogenomics, respectively (see Chapter 8). (vi) Bioinformatics: It is a new field of biotechnology linked with information technology. Bioinformatics may be defined as application of information sciences (mathematics, statistics, and computer sciences) to increase the understanding of biology, biochemistry and biological data. The most remarkable success of bioinformatics to date has been its use in the 'shotgun sequencing' (breaking of a large piece of DNA into smaller fragments) of human genome. D. BIOTECHNOLOGY IN INDIA AND GLOBAL TRENDS 1. Biotechnology in India The recombinant DNA technology has become the major thrusts in most of the developing countries. In 1982, Government of India set up an official agency, the National Biotechnology Board' (NBTB) which started functioning under the Department of Science and Technology (DST). In 1986, NBTB was replaced with a full-fledged department, the Department of Biotechnology (DBT), under the Ministry of Science and Technology for planning, promotion and coordination of various biotechnological programmes. The DBT is making effort in promoting post graduate education and research. Special M.Sc. courses in Biotechnology in selected group of institution with scholarship are provided by the DBT. The selection of students is done via National Test. In addition, it also provides trained manpower for the rapidly growing biotech industry. It has also raised the level of biology education in certain areas of biotechnology in the country. Moreover, a considerable amount of basic biochemical and molecular biology is imported in these courses. Copyrignted material  Biotechnology: Scope and Importance 13 India has the DBT, DST, CSIR, ICAR, ICMR and IARI, and other agencies which are working under the Government. These agencies and the other National and International Industries are man ufacturing Biotech products and marketing them after clinical trials. A Technology Development Board (TDB) has been set up by the Government for the promotion of product development. The TDB works with universities, industries and the National Institutes. The Technology Information, Forecasting and Assessment Council (TIFAC) has prepared a Vision 2020° document which consists of biotechnology also. Since 1980s, India has supported a lot to biotechnology industry and its products. Teaching and research ofbiotechnology have been included in University's syllabi both at Under Graduate and Post graduate levels. DBT- supported departments are running in several Universities and Institutions. It is hoped that India will play a key role in future as one of the largest market of the world, and as a producer of biotech products. () International Centre for Genetic Engineering and Biotechnology (1CGEB). The United Nations Industrial Development Organisation (UNIDO) recognised the potential of genetic engineering and biotechnology for promoting the economic progress of the developing countries. The initiation taken by UNIDO has led to the foundation of 1CGEB. In 1981, in a meeting convened by UNIDO it was proposed to establish an international centre of excellence to foster biotechnology in the developing world. In 1982, this concept was approved by a high level conference of developed and developing nations in Belgrade. The statutes of the centre were signed by 26 countries with the entry into force of statutes on February 3, 1994. The lCGEB has become a fully autonomous international organisation composing of at present 33 member states. The ICGEB has its two centres, one located in Trieste (Italy), and the other in Jawaharlal Nehru University, New Delhi (India). The Trieste component is currently occupying about 5,700 m area, whereas the New Delhi component is occupying about 10,000 m² area. This centre is functioning in a proper way since 1982. The organs of ICGEB are the Secretariat, the Board of Governors and the Council of Scientific Advisors. The secretariat component is the Director, two Heads of the components and the scientific and administrative staff operating with the framework of the ICGEB programme. The Board of Governors consists of a representative of each Member State. The Council of Scientific Advisors is composed of eminent scientists and overseas scientific excellence of ICGEB. Funds are provided by the government of Italy and India. From 1999, all Member States have started to finance ICGEB through a scale of assessment adopted by the Board of Governors. The activities of ICGEB are aimed specifically at strengthening the R & D capability of its Member States by : (1) providing the developing countries with a necessary 'critical mass' environment to pursue and advance the research in biotechnology; host research facilities that are technology and capital demanding and, therefore, inaccessible to the great majority of developing countries, (ii) training schemes and collaborative research with affiliated centres to ensure that significant members of scientists from Member states are trained in state-of-art technology, in areas of direct relevance to the specific problems of their countries, and (iii) acting as the coordinating hub of network of affliated centres that serve as localized nodes for distribution of information and resources located at ICGEB. Initially a total of six centres were set up at various Universities/Institution namely, Jawaharlal Nehru University (New Delhi), Madurai Kamraj university (Madurai), Tamil Nadu Agriculture University (Coimbatore), National Botanical Institute (Lucknow), and Bose Institute (Kolkata). In addition, the other centres for biotechnology in India are: Indian Agricultural Research Institute (LARI), Jawaharlal Nehru University (JNU), Delhi University, Indian Veterinary Research Institute Copyrigted materal  14 A Textbook of Biotechnology (IVRI), Izzatnagar (U.P.); Central Food and Technology Research Institute (CFTRI), Mysore; Na tional Dairy Research Institute (NDRI), Karnal (Haryana); Malaria Research Centre (MRC), Delhi; Regional Research Laboratory (RRL) Jammu; Central Drug Research Institute (CDRI) and Central Institute of Medicinal and Aromatic Plants (CIMAP), Lucknow; Indian nstitute ofTechnology (IIT), Kanpur, Madras, Bombay and New Delhi. Other centres to which DBT has provided infrastructural facilities are Banaras Hindu University, Varanasi; Allahabad University; M.K. University, Madurai; Anna University (Madras); Indian Institute of Science (Bangalore); Pune University, Pune; AIl India Institute of Medical Sciences, New Delhi; Bhabha Atomic Research Centre (BARC), Bombay, etc. Mr. Rajiv Gandhi, the Late Prime Minister of India, laid a foundation stone on October 4, 1988 at the Centre I.A.R.I. with the name Lal Bahadur Centre for Biotechnology". Many public and private institutions working under the Government departments and organisations have advised the DBT to formulate the biotechnology programmes under the following areas () plant molecular biology and agricultural biotechnology, (ü) biochemical engineering, process optimisation and bioconversion, (iüi) aquaculture and marine biotechnology, (iv) fuel, fodder, biomass and green cover, (v) medical biotechnology, (vi) microbial and industrial biotechno logy, (vii) large scale use of biotechnology, (viii) integrated systems in biotechnology, (ix) Veterinary biotechnology and () Infrastructural facilities. In India, the phamaceutical industry is very strong and vibrant with expertise for chemical drugs. It has little experience in biotech diagnostics and no experience in biotech therapeutics. Moreover, pharma industry is located between Mumbai and Ahmedabad (90% of drug production in India is in Gujarat and Maharashtra). There is no Government institution or university with expertise in this area to help pharma industry. However, for a variety of reasons, the Indian pharmaceutical industry will sooner or later enter in manufacturing of biotechnology-based diagnostics and therapeutics. (iü) Needs for Future Development. A few developing countries like India, have scientists and technologists related to biotechnology where national strategies of development in biotechnology could be implemented. The scientific and technical manpower has to be properly shifted towards new biotechnology with the aim to produce expertise in biotechnology. In a keynote address, Bachhawat and Banerjee (1985) have described the impact of biotechnology on third world countries. They emphasized "Indian bioscientist must be trained to utilize their knowledge and expertise for application and orientation, for example, a microbiologist must be trained in microbial genetics to be really useful in fermentation technology, or a botanist must be trained in cell culture, protoplast fusion or DNA recombination for practical utility and similarly, people from traditional disciplines in life science may be trained to reorient their knowledge towards application and process of training readjusted according to need." In India, most of the universities have started teaching biotechonology at under-graduate level. However, at post graduate level teaching and research have been initiated only by a few universities/ institutes on all India entrance test basis. Government of India has selected many thrust areas of national and international relevance as described earlier. 2. Global Scenario Day-by-day biotechnology products are increasing in world market. The high value added biotechnology product to be used in medical field are now in domination for the last few years. The countries which have boosted up biotechnology R& D during the past two decades are the U.S.A., U.K., Japan, France, Australia, Russia, Poland, Germany, as well as India (among the developing countries). The most effective means of promoting international cooperation is through networks. The Copyrignted matcrial  Biotechnology : Scope and Importance 15 international networks devoted to applied microbiology/biotechnology are the Regional Microbiology Network for South-East Asia, and the network of Microbiological Resource Centres (MICRENs). To foster biotechnology inventions, the U.S.A. promotes enterprises through policy development and support. Funding for basic scientific research at the National Institute of Health (NIH) has been supported. The U.S. administration has boosted up the process for improving new medicines so that these may be quickly and safely available in market. The private sector research investment and small business development have also been encouraged through the incentives. Position of therapeutic proteins and vaccines up to June 1998 by the USA has been given in Table 1.6. > Table 1.6. Position of therapeutic proteins and vaccines up to June 1998 (based on Phamaceutical Research nd Manufacturers of America; Biotechnology Industry Organisation). Therapeutic proteins/vaccines Status Approved In development Blood clotting factor 3 3 Gene therapy 38 Growth factors 21 Human growth hormones 5 Inteferons 7 9 Interleukins 9 Monoclonal antibodies 12 72 Recombinant human proteins 6 4 Vaccines 5 77 Others 12 52 The international markets have been opened for biotechnology research and biotech products. One of the world's best examples of partnership has becen established by developing public databas es. It enables the scientists to tie up with an enterprise. This has helped in developing partnership among University research-Government and private industries. Science education has been improved. Guidelines have been prepared so that science based regulatory programmes: () can promote public biosafety, (ii) earn public confidence, and (iii) can guarantee fair and open international market. The other countries (such as U.K., Japan, Germany, France, Australia, etc) have also prepared similar guidelines for promotionof biotechnology products and bio-business, E. POTENTIAL OF MODERN BIOTECHNOLOGY The modern biotechnology is expected to solve many problems arising at the global level. In 21st century growth and economy of acountry will certainly depend on operation of biotechnology. A revolution may occur in some of the areas like medical and health care, agriculture, industry and environment. DNA fingerprinting has successfully helped the forensic science in the search of criminals mak ing identity of individuals, solving parentage dispute, etc. Human diagnostics and therapeutic drugs have been discovered and commercialised. Gene therapy is hoped to solve the problems of genetic diseases. Biotechnology-based vaccines are the best as compared to conventional vaccines, and so is the insulin. They have no side effects and pose no risk for the presence of live form of viruses in vaccines. Many transgenic plants and animals have been genetically improved. Now they are capable of producing new or improved products. The questions may be raised on complex, ethical, spiritual and philosophical issues. Copyrigted materal  16 A Textbook of Biotechnology Hundreds of transgenic plants (a) (b) have been produced and many of them are being sown in field and products are available in market. The plant biotechnology will reduce the dependence of farmers on pesticides and will help to utilise the new tech nologies. In near future papers and chemicals with less energy and less pollution may be produced through biotechnology. Abatement of pollution us ing potential microorganisms (i.e. bioremediation) has been used in many countries. Thus the potential (a) Buds of cotton plants are vulnerable to worm attack microorganisms are looked for better (b) Buds of a modified cotton plant-resist attack. health, better products and better environment of future. Instead of chemical pesticides, biopesticides have been produced and com mercialised by some industries in India and abroad. Similarly, biofertilizers (formulation of nitrogen fixing bacteria and blue green algae, i.e. cyanobacteria, and phosphate-solubilising bacteria and fungi) have also been made available to farmers in India. On the other hand, biotechnology has generated new jobs for the youth and stimulated the growth of small business. It has also encouraged the innovations in the industrial and agricultural sectors. In the U.S.A. alone more than 15 lakh youth have been employed in industries. E. ACHIEVEMENT OF BIOTECHNOLOGY In recent years, it has become possible to map the whole genome of an organism to find out the function of the genes, cut and transfer into another organism (see Chapters 3 and 4). Owing to the success achieved from gene cloning, many products have been obtained through genetically engineered cells, and hopefully many can be produced during the current decade. Recombinant DNA technology has made it easier to detect the genetic diseases and cure before the birth of a child or suggest accordingly. Gene bank and DNA clone bank have been constructed to make available different types of genes of its known function. Thus, recombinant DNA technology has made it possible to develop vaccines against viral and malarial diseases, growth hormones and interferon (see Chapter 7). Biotechnology has caused revolution in agricultural science. Cell culture and protoplast fusion techniques have resulted in hybrid/cybrid plants through inter generic crosses which generally are not possible through the conventional hybridization techniques (see Chapter 12). It has also helped in the production of encapsulated seeds, somaclonal variants, disease resistant plants, Transgenic plant developed by recombinant DNA technology. herbicide-and stress-resistant plants, and nif gene and Copynghted matenat  Biotechnology : Scope and Importance 17 nod gene transfer as well. Through cell culture techniques, industrial production of essential oils, alkaloids, pigments, etc. have been boosted up. However, many more works are to be done on horticulture and forestry plants as far as micropropagation and establishment of mycorrhizal fungi are concerned (see Chapter 13). For better yield of agricultural crops, use of biofertilizers (seed bacterization, algalization and green manuring) has become an alternative tool for synthetic chemical fertilzers. The biofertilizers are non-toxic to micro- and macro-biota and to humans as well. This would reduce the constrain on fossil-fuel based industries (see Chapters 19 and 20). Moreover, to discourage the use of synthetic pesticides, biocontrol agents have been developed and conditions have been investigated when phenomenon of antagonism takes place (see Chapter 21). For the protection of environment and abatement of pollution, treatment of sewage, transformation of domestic wastes and xenobiotic chemicals have drawn much attention in recent years. To combat these problems such bacterial plasmids have been developed that could be used to degrade the complex polymers into non-toxic forms. Strains of cyanobacteria, green algae and fungi have been developed which could be used for the treatment of municipal and domestic sewage and industrial discharges into nontoxic forms and renew them as source of energy. Biotechnology has helped the bio-industries in producing the novel compounds and optimization, and scale up of products, for example alcohols, acids, antibiotics and enzymes (see Chapters 17 and 22) and single cell protein and mnycoprotein (see Chapter 18). Technologies have also been developed to seek an alternative source of energy from biomaterials generated from agricultural, industrial, forestry and municipal sources (see Chapter 23). Social forestry and short rotation tree plantation will help to reduce the pressure on forests to meet the demand of fuel in rural sector. In industries, biomass fired system have been developed to meet the energy requirement of engines, such as sugar cane mills. Moreover, urban sewage and plant weeds are used for the production of biogas for cooking and lighting purposes (see Chapter 24). 1. Ban on Genetic Food It is a growing concern all over the world that the genetic food may pose risks to human health, ecology and the environment. However, it has forced the government of many countries to re-think on introduction of such designer crop. For the first time the European Commission's Scientific Advisors have recommended that a genetically engineered potato be withheld from the market because they cannot guarantee its safety. Worried at the growing acts of vandalism against the genetically engineered crops in Britain, the environment minister had gone on recod that his government was considering a three year ban on transgenic crops grown for commercial use. The United States, the world's biggest producers of genetically modified foods, has also threatened New Zealand to ban his genetically engineered foods. In Europe, the boom in the stock market for biotech products also is in wane. British Biotech, Europe's Flagship Biotechnology Company has lost its share value by more than fifty percent. However, it is a general opinion that in solving the global problems of hunger and food shortages biotechnology cannot make food cheaper at any cost. It is true that cotton pests, especially American bollworm has become resistant to certain pesticides. Moreover, several pests have developed immunity against the Bt gene (Bacillus thuringiensis gene, see Chapter 21). If the cotton bollworm too develops resistance against the Bt-cotton, willforce a still large number of farmers to commit suicide as happened in Andhra Pradesh in 1998 and in subsequent yearS as in Gujrat in 2002. Copyrignted materal  18 A Textbook of Biotechnology F. PREVENTION OF MISUSE OF BIOTECHNOLOGY In many countries the prevention of misuse of biotechnology is stressed because it may cause a lot of mischief when given a free play in the hands of transnational companies engaged in ruthless persuit of profits. One of these concerns is the rapid pace of genetic erosion. This will lead to a situation where the base genetic material is available to only a few multinational companies in their gene banks. However, this genetic erosion has to be checked by saving the genetic diversity in its own environment, but not gene bank/germplasm bank, by the involvement of people. We are proceeding from 'green revolution' to 'gene revolution'. New transgenic plants and animals are being produced, and seeds, embryos, sperms are preserved. A day may come when farmers would have to depend on genetically engineered seeds. Still, there is doubt whether such seeds will suit for sustainable agricultural practice free from chemical poison. The genetically modified organisms (GMOs) should be carefully researched and monitored to ensure that the hazards to users and environment will not occur. In addition, concerted action should be undertaken to ensure that necessary consideration is given to the ethical and social effects of such studies. People should be known about the impacts of GMOs and genetically engineered products. Such efforts are already being made in some of the countries. The German Green party has called for a 5-year moratorium on commercial release of GMOs. The UK genetics forum is complaining for a partial mentally iresponsible applications of biotechnology. In the USA, a number of groups are strongly opposing the deliberate release of GMOs. In India, gene campaign has been carrying out a public campaign against the patenting of life forms and the misuse of biotechnology. However, such efforts need to be strengthened to check the misuses ofbiotechnology. G. BIODIVERSITY AND ITS CONSERVATIONS Biodiversity, is a new name for species-richness (of plants, animals and microorganisms) occurring as an interacting systen in a given habitat. Biodiversity cannot be replaced because the species becomes adapted in a given habitat after a long course of time. That is why, due to plausticity in their nature and unsustainable resource utilization, over 2.5 lakh species are lost and thousands are threatened to extinction. If a species extincts, it means whole of the gene pool extincts. The real value of biodiversity lies in the informations that are enclosed in the genes. Therefore, there is urgent need, for future, to protect the genes from destruction. Biodiversity may be defined as the inherent and externally imposed variability within and among the living organisms present in terrestrial, marine and other ecosystems at the specific time. Diver sity includes the variability in genes, genotype, species, genera, family and ecosystem at a particular time in a specific region. Thus, biodiversity is an expression of both numbers and differences and differences can be seen as a measure of complexity. Commonly biodiversity may be considered at the following three different levels: () Genetic Diversity: Genes are the functional entities of all organisms because they determine the physical and biological features of organisms. Genes of one organism differ from that ofthe other organisms. Besides, variations arise due to mutation brought about in genome which is the cause of genetic diversity. (ii) Species Diversity: A species is a group of organisms which are genetically identical and interbreed to produce progeny. In contrast horses and zebra are two different species but genetically similar. They can interbreed but produce all infertile offspring. Usually species differ in appearance and thus one can differentiate one species from the other. Thus species diversity is estimated on the basis of total number of species within the discrete geographical boundaries. (iii) Ecosystem Diversity: There is diverse ecosystems, and organisms living in such ecosys Copyrighted materal  Biotechnology : Scope and Importance 19 tems are adapted to its respective ecosystems. Therefore, there arises diversity among them. Under ecosystenm diversity two phenomenon are frequently referred: the variety of species within different ecosystems i.e. more diverse ecosystems contain more species, and the variety of ecosystems found within a certain bio-geographical boundaries. 1.Biodiversity in India India is rich in biodiversity to agriculture, animal husbandry, fisheries and forestry. Much has been described in Ayurved and other ancient literature about the indigenous system of medicine, knowledge and wisdom of people. These are supported by a very strong scientific and technological base. In India, over 1,15,000 species of plants and animals have already been identified and described. The country is an important Centre of Diversity and origin of over 167 important cultivated plant spices and domestic animals. A few crops which arose in India and spread throughout the world are : rice, sugarcane, vignas, jute, mango, citrus, banana, millets, spices, medicinal aromatics and ornamentals. No country in the world is as rich in biodiversity as ours. Himalaya itself has the natural wealth of plants, many of which are still unknown and many endangered. 2. Conservation of Biodiversity There is an urgent need for biodiversity rich countries to save it against destruction. However, in most of the developing countries, biodiverisity is attached to environment and forest agencies which have no idea about it. If such countries are not aware of conserving it for sustainable utilization, they would be compelled to export biodiversity and import products for well being of their people. The agreement between Institute of Biodiversity (INBio) in Costa Rica and Merck (USA) is hailed throughout the industrial world. Under this agreement, extracts, from the wild plants, insects and microorganisms from Costa Rica are supplied to Merck. In return, INBio receives from Merck over l1.35 million US dollars, and expects royalty on the commercial products. INBio has to contribute 50% of royalty to the Government of Costa Rica for National Park Service. The Government has given to INBio rights to bioprospects and share conservation work. Thus, the INBio represents an alliance between biologists/ biochemists and businessmen. In India, a large number of institutions are involved in conservation and utilization of biodiversity which come under Ministry of Environment and Forest, Agriculture, and Science and Technology. They deal conservation of biosphere reserve, national parks, wild life, sanctuaries, field gene banks, etc. The country needs more expertise and methodologies besides tiger-bird wild life syndome. India is predominantly an agricultural country. Therefore, the policy makers have to realize that conservation and sustainable utilization of biodiversity must be placed on the top of all developmental plannings. H. GENE BANK (GERMPLASM BANK) AND PLANT CONSERVATION Out of 2,50,000 different plant species, some are already lost and nearly 20,000 seed plants are threatened to extinction. Due to disappearing of natural resources at fast rate, genetic variabilities are being lost for ever, which will result in a dangerous future. Therefore, to save the threatened and gradually vanishing species, and to meet the world demand of food, we would have to conserve the natural heritage. The gene bank (germplasm bank) has taken the challenge of conserving the gene pool of economically, medicinally, socially and ecologically important plant species. In the first decade of the 20th century a Russian scientist, N.I. Vavilov, was the first to realize the need of conservation of plant genetic resources. Later, in 1920, he established the first genetic resource centre (GRC) of the world at Leningrad. Thereafter, a global concern was raised after the International Biological Programme in 1964. Copyrignted matcrial  20 A Textbook of Biotechnology 1. World Genetic Resource Centres During the last two decades, many regional and international GRCs have been set up in different countries. International Rice Research Institute, Manila (Philippines) has rice germplasm bank where 25,000 varieties of rice germplasm has been collected throughout the world. Similarly, maize germplasm have been stored at Maize and Wheat Improvement Centre, Mexico (with more than 12,000 varieties), Institute Colombiano Agropecuario, Colombia (more than 2000 varieties), Instituto National de Investigations Agricolas, Mexico (with more than 7000 varieties), and potato germplasm at International Potato Centre, Lima (Peru). A seed bank has been set up at the National Bureau of Plant Genetic Resources (NBPGR) (New Delhi) which is associated with a world network of gene resource centres coordinated by the International Bureau of Plant Genetic Resources of FAO. 2. Conservation and Storage For the conservation of genetic materials, seeds, pollen grains, vegetatively propagating parts, plant tissues or genome of plant cells are collected. These depend on: (1) ease to handle, (ii) nature of crops, (ii) longevity, (iv) expertise, (v) methodologies available, (vi) space required, (vi) problems of genetic erosion of stocks, and (vii) expenses. The seeds are compact and easy to handle; hence seed collection and preservation are commonly adopted. Seeds are stored in sealed hermetic containers, dried to a desired moisture level and maintained at desired low temperature. Viability of pollen grains differs in different species; the species having maximum viability are selected. The families showing the highest longevity of pollen grains are: Pinaceae, Primulaceae, Rosaceae and Saxifragaceae. The longest period of pollen storage recorded is 9 years at -20°C for apples. The vegetatively propagating parts (corn, bulb, tubers, etc) of some plants such as Dioscorea, potato, sweet potato, etc. are used for storage purposes. Techniques have been developed to culture the plant cell, tissue and organs. In vitro grown cultures (e.g. plantlet, apical meristem culture, etc.) are stored (see Chapter 10). Storage of in vitro grown cultures has many advantages over the others such as: (i) requirement of less space, (ii) cheap in maintenance, (ii) high propagation potential, (iv) least problem of genetic erosion of stock, and (v) maintenance of pathogen free stock. PROBLEMS 1. What is biotechnology? Discuss brief the different areas of interest. 2, Why can microbiology, genetics and biochemistry, without combining with technology, not be called as biotechnology? 3 What are the achievements of biotechnology? 4. Write an essay on global impact of biotechnology. 5. Write an essay on History of biotechnology. 6. Discuss in detail how did traditional biotechnology help in the evolution of biotechnology? 7. Give a brief account of emergence of modern biotechnology 8. Give a brief note on potential of modern biotechnology 9. Write short notes on the following : (i) NBTB (ii) DBT (üi) OECD (iv) ICGEB (v) Lal Bahadur Shastri Centre for Biotechnology (vi) Biotechnology in India, (vii) Biodiversity (viii) Ban on genetic food. (ix) Modern biotechnology (x) Traditional biotechnology Copyrignted material